Adhesive functional requirements for common transdermal patch designs.
\r\n\t
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Enzymes are one of those molecules that are globally recognized for their multifarious applications in industries. For instance, their utility in brewing, dairy products, detergents, food and feed, pharmaceutical production, and paper and pulp industry is huge. One of those most widely used enzymes is cellulase. According to recent global cellulase market analysis reports, the demand for this enzyme is exponentially increasing.
\nCellulose, the substrate of cellulase, is the most abundant polysaccharide present on earth. It is the main substance in plant materials. Anselme Payne was the very first person to discover and isolate this amazing compound from green plants [1]. It happened more than two centuries ago. From the past, cellulosic materials have played a crucial role in daily human life. They used it to fertilize their soil for crop cultivation. It was also fodder for their cattle. It was firewood for cooking, and they were igniting cellulosic material to generate heat whenever they needed to produce energy.
\nCurrently, the role played by cellulose is not that simple. Especially, as it is recognized as a cost-effective raw material, the useful applications of cellulose in the industrial sector have become much more complex. This has laid a huge platform for scientists to do cellulose-based research in multidisciplinary approaches. One such area is hydrolysis of cellulose. In nature, this is usually accomplished by cellulases. Cellulase is catalyzing hydrolysis of cellulose.
\nHowever, cellulase is not a single enzyme. It is a group of enzymes which is mainly composed of endoglucanase and exoglucanases including cellobiohydrolases and β-glucosidase. Fungi, bacteria, and actinomycetes are recorded to be efficient cellulase enzyme producers in the natural environment. These microorganisms must secrete cellulases that are either free or cell surface bound. Their enzyme production efficiency and the enzyme complex composition are always diverse from each other. Although both aerobic and anaerobic microorganisms produce these enzymes, aerobic cellulolytic fungi, viz.,
According to recent enzyme market reports, the key areas of the industry where cellulase enzyme is increasingly being applied are healthcare, textile, pulp and paper, detergent, food, and beverages. Its wide application in coffee processing, wine making, and fruit juice production is related to food and beverage segment. In other industrial applications, it is broadly used to produce laundry detergents and cleaning and washing agents. Cellulase is also being highly recognized as an effective alternative to available antibiotics for treatment of biofilms produced by
Application of microorganisms or microbial enzymes for pretreatment of lignocellulosic material is currently earning a huge attention of the industry. This is a result of growing interest about depletion of fossil fuel resources in the world which have inspired the production of bioethanol from lignocellulosic biomass through enzymatic hydrolysis [3]. Lignocellulosic biomass is one of the best options as a low-cost, readily available, eco-friendly raw material. However, it is not found alone. Cellulose is forming lignocellulose in combination with hemicellulose and lignin which finally becomes a compact network structure [4]. Moreover, it has a crystalline structure which is hard to break down. Therefore, cellulose is insoluble in water and causes limitations in hydrolysis. That is why it is essential to pretreat lignocellulosic material in industries like bioethanol production. During pretreatment, it will loosen up the crystalline structure and facilitate the degradability to release fermentable sugar forms. There are several methods available for pretreatment of lignocellulose, viz., physical, chemical, and biological methods. Biological pretreatment using cellulolytic microorganisms and their enzymes is found to be the best way of addressing this problem.
\nBy all means, cellulase is an enzyme which can cause a huge economic impact. However, there are some considerable bottlenecks of utilizing this enzyme in the industry. For example, the higher cost of cellulase and less catalytic efficiency are especially understood. Another important point is less understanding of the relationship between hydrolysis mechanisms and molecular structure of the enzyme. This knowledge is important to carry out further improvements in the enzyme to enhance its catalytic activity. Therefore, this chapter is discussing about the structure and function of cellulase in order to understand its mechanisms of action. The details on current applications of the enzyme have also been summarized here. Furthermore, the efforts have been taken to bring together information on novel biotechnological trends of cellulase. Moreover, it is discussing about possible low-cost, enzymatic pretreatment methods that have been practiced for lignocellulosic materials in order to use it as an efficient raw material to produce bioethanol.
\nBefore moving on to cellulase, it is essential to understand cellulose which is the substrate of cellulase enzyme. This section will provide a short description about cellulose.
\nCellulose is a linear polysaccharide. In this polymer, D-glucose subunits are attached together by formation of β-1,4-glycosidic linkages between individual glucose molecules. The molecular formula of cellulose is (C6H12O6)n. The “n” indicates the degree of polymerization (DP). It symbolizes the number of glucose subunits connected with each other. This number is varying from hundreds to thousands. Two glucose repeating units together are called cellobiose. In other words, this polymer is made by β-(1 → 4)-D-glucopyranose units in 4C1 conformation. It consists of long chains of anhydro-D-glucopyranose units (AGU) with each cellulose molecule having three hydroxyl groups per AGU with the exception of the terminal ends. Cellulose has both crystalline and amorphous regions in its structure in various proportions [5]. Those regions are intertwined to form the structure of cellulose. There are four major crystalline forms, for instance, Iα, Iβ, II, and III. This crystalline structure is a result of intramolecular and intermolecular hydrogen bonding between glucose monomers in cellulose. These hydrogen bonds construct a huge network that directly contributes to the compact crystal structure of cellulose polymer. On the other hand, this strong intramolecular and intermolecular hydrogen bond formation leads to poor solubility of cellulose.
\nIn plant cell walls, cellulose exists as different levels of structures, i.e., single cellulose chains, elementary fibrils (consisting of tens of single cellulose chains), and microfibrils (bundles of elementary fibrils). It is proposed that the macrofibril is composed by the attachment of several newly synthesized elementary fibrils. With the cellular growth, the macrofibrils divide to form individual microfibrils. Microfibril is consisting of a single elementary fibril. Although elementary fibrils and macrofibrils are composed of mere cellulose, microfibril has noncellulosic polymers like hemicelluloses along with cellulose. It is noted that others consider a microfibril as consisting of a number of elementary fibrils. Microfibril is an elementary fibril associated with noncellulosic polymers. Each microfibril might contain up to 40 cellulose chains and is about ~10 to 20 nm in diameter. Many such cellulose chains aggregate into bundles called micelles and micelles into microfibrils. Micelles are interconnected with few cellulose fibers. The plant cell wall structure is stabilized by the macrofibrils. The cross-links between hemicellulose and pectin matrices also support this stabilization process. Lignin is a complex polymer which usually fills the spaces between cellulose and pectin matrices. It forms covalent bonds with hemicellulose. This provides more mechanical strength to the plant cell wall. This structure which is present in plants is collectively called lignocellulose. Other components known as extractives including fats, phenolic, resins, and minerals are also present in lignocellulosic biomass.
\nEnzymes are known to be very useful in many industrial processes. Their broad applicability has created a significant market demand in the recent years. According to market reports on world enzyme demand (2017), they have recognized several key factors which lead to huge consumer demand for enzymes. Some of them are completely bound with economical advances. For example, increased per capita income in developing countries causes huge growth in consumer-related industrial applications [6]. A recent industry study done by Freedonia in January 2018 on “Global Industrial Enzymes” reveals that global demand for industrial enzymes is projected to grow 4.0% per year to $5.0 billion in 2021. This report also emphasizes the gains in personal incomes in developing countries as the key factor which is supporting growth in demand for enzymes. The development of scientific research on enzymes is mainly based on disciplines such as biotechnology,molecular biology and genetics. Continued advances in these areas of research, particularly related to DNA manipulation and sequencing, result in extensive increases in enzyme demand worldwide. Cellulase is one such enzyme which earns consecutively increasing demand. Therefore, collection of knowledge about this enzyme is essential for further development of fundamental and applied research on cellulase and for consequent application in human life.
\nIt is produced by fungi, bacteria, actinomycetes, protozoans, plants, and animals. According to Carbohydrate-Active Enzymes database, there is information of the glycoside hydrolase families. Glycoside hydrolases, including cellulase, have been classified into 115 families based on amino acid sequence similarities and crystal structures. A large number of cellulase genes have now been cloned and characterized. They are found in 13 different families. Furthermore, there are 3D structures of more than 50 cellulases. All of cellulases cleave β-1,4-glucosidic bonds. However, they display a variety of topologies ranging from all β-sheet proteins to β/α-barrels to all α-helical protein.
\nIn the structure of cellulase, there are catalytic modules and non-catalytic modules. The catalytic modules of cellulases have been classified into numerous families based on their amino acid sequences and crystal structures. The non-catalytic carbohydrate-binding modules (CBMs) and/or other functionally known or unknown modules may be located at the N- or C-terminus of a catalytic module. Usually, fungal and bacterial cellulase mainly has two or more structural and functional domains. Both aerobic and anaerobic microorganisms are producing this enzyme. Therefore, there are two types of cellulase systems: noncomplex and complex. A noncomplex cellulase system is produced by aerobic cellulolytic microorganisms, and it is a mixture of extracellular cooperative enzymes. In a noncomplex cellulase system, the common arrangement is joining of a catalytic domain with a cellulose-binding domain (CBD). A complex cellulase system is produced by anaerobic microorganisms and it is called “cellulosome.” Cellulosome is assembled by joining a catalytic domain with a dockerin domain. The enzyme is a multiprotein complex anchored on the surface of the bacterium by non-catalytic proteins that serves to function like the individual noncomplex cellulases but is in one unit.
\nIn addition to these two major domains in the cellulase structure, there are some other domains that are present in many cellulases, for instance, S-layer homologous (SLH) domain, fibronectin-type 111 domains, and NodB-like domain, and there are also other regions of unknown function. These domains are often connected by Pro and hydroxy amino acid (threonine and serine) enriched linker sequences. Among all these domains, catalytic and cellulose-binding domains are the most important because they are the domains which are considered participating in hydrolytic mechanisms of the enzyme.
\nCellulase catalyzes the decomposition of cellulose polysaccharide by simply breaking down β-1,4-glycosidic bonds. Three major types of enzymes are generally involved in hydrolyzing cellulose microfibrils in the plant cell wall: endoglucanase, exoglucanase, and β-glucosidase. Complete cellulose hydrolysis is mediated by the combination of these three main types of enzymes. Endoglucanase usually attacks amorphous areas of cellulose. The random attack of this enzyme on internal bonds of loosely bound, amorphous areas of cellulose creates new chain ends. These new chain ends are then easily attacked by other types of enzymes. The highest activity of this enzyme usually occurs against soluble cellulose forms or acid-treated amorphous cellulose. The function of exoglucanase is to produce glucose or cellobiose units by attacking the reducing or nonreducing end of cellulose chains. Endoglucanase is different from exoglucanase because it is usually very active against crystalline cellulose substrates such as Avicel or cellooligosaccharides. Finally, β-glucosidase can hydrolyze cellobiose to glucose from the nonreducing ends, and it is inactive against amorphous or crystalline cellulose. Although an exact mechanism is not yet finalized, fragmentation of cellulose aggregations into short fibers has been observed and reported during the beginning of cellulose hydrolysis prior to releasing any detectable amount of reducing sugars. This is known as amorphogenesis.
\nThere are two catalytic mechanisms of cellulases. They are simply introduced as retaining mechanisms and inverting mechanisms. Cellulases cleave glucosidic bonds by using acid-based catalysis. The hydrolysis is performed by two catalytic residues of the enzyme: a general acid (proton donor) and a nucleophile/base. The catalytic mechanism which occurs depends on the spatial position of the catalytic residues. The retention and inversion of the anomeric configuration of cellulose are the two mechanisms which hydrolyze cellulose. The “retaining” cellulases retain the same configuration of anomeric C bearing the target glucosidic bond even after a double-displacement hydrolysis with two key glycosylation or deglycosylation steps. “Inverting” cellulases inverts the configuration of the anomeric C configuration after a single nucleophilic displacement hydrolysis [7].
\nFor many decades, cellulases have played a crucial role as biocatalysts. They have shown their potential application in a large number of industries. Textile, paper and pulp, laundry and detergent, agriculture, medicine, and food and feed industries are some of the major industries which employ microbial cellulases. According to Coherent Market Insights, the textile industry is the dominant market for cellulases in 2017. According to most of the enzyme market research reports published in 2018, food and beverages, textile industry, animal feed, and biofuels have been reported to be the major areas of applications.
\nAccording to another Global Cellulase (CAS 9012-54-8) Market Research Report published in 2018, Asia-Pacific is the largest consumer of cellulase, with a revenue market share nearly 32.84% by 2016. Furthermore, the reported data showed 29.71% of the cellulase market demand in animal feed, 26.37% in food and beverages, and 13.77% in the textile industry in 2016. This same report forecasts that the applications of cellulases will reach 2300 million USD by the end of 2025, growing at a compound annual growth rate (CAGR) of 5.5% during the 2018–2025 period. These data suggest that the application of cellulases in industries is drastically rising annually. Novozymes and DuPont from Denmark are key cellulase enzyme producers supplying these enzymes to the global market for industrial applications. From this point forward, in this chapter, our major effort was to discuss about the current applications of cellulases in major fields that have been listed above. The novel biotechnological trends emerging in those fields while understanding the key areas of research where further studies required also surfaced to an extent.
\nThe textile industry is one of the largest industries in the world. The customer demand for fashion is increasing as they want uniqueness in styles, colors, and the clothes they wear. There was a significant growth in this industry during the last few decades as a result of this increasing customer demand. This enzyme has now become the third largest group of enzymes used in these applications [8]. This creates a very competitive market platform for manufacturers that are always looking for environmentally friendly approaches of giving their products a unique look. Cellulase is used for many purposes in the industrial sector.
\nEspecially for textile wet processing, biostoning of denim fabric, biopolishing of textile fibers, softening of garments, and removal of excess dye from the fabrics are some of the major applications of this enzyme in the industry. Fungal cellulases from
Biostoning and biopolishing are well known for the best applications of cellulases in the current textile industry.
\nThe conventional washing process of denim usually has three steps. The denim fabric is first treated with amylase enzyme to remove the starch coating of the fabric. This process is called desizing. During this process, starch is broken down into maltose which is a water-soluble disaccharide composed of two glucose molecules. Then, the fabric is given treatment by providing abrasion to the material in pumice stones added to the washing machine. This wash was completely achieved by adding chemicals like sodium hypochlorite or potassium permanganate. This traditional process has several disadvantages. The addition of pumice stones must be done in larger quantities. This was affecting the machine’s productivity in an adverse way causing tear effects. After the wash is completed, the manual removal of stones is needed. This is causing further reduction of the process efficiency. The excessive back-staining was another disadvantage of the traditional process. Backstaining is the reaction by which the removed dye molecules are deposited on the denim fabric again.
\nApplication of microbial cellulases was found to be an efficient alternative for pumice stone washing. It was first staged in the 1980s. The use of stones is currently replaced by cellulases in a successful way. During this process, cellulases act on the denim fabric which is made of tough cotton. The indigo dye which is used to color the fabric is trapped inside the cellulose fiber in this cotton material. Usually, the indigo dye is mostly attached to the surface of the yarn and to the most exterior short cotton fibers. When the fabric is treated with the enzyme, it hydrolyzes and breaks small fibers coming out of the fabric which loosens the dye. For this purpose, the β-1,4-linkages of cellulose chains will be broken down, and simple water-soluble sugars will be formed. This will remove the fibers which traps indigo dye. Then, the dye is easily removed from the fabric giving a faded look.
\nThe major disadvantage associated with the application of microbial cellulases is again backstaining. The redisposition of dye on the fabric covers up the shaded look given by the treatment. In order to overcome this problem, several biotechnological approaches have been already experimented by researchers. Immobilization of cellulases on pumice stones is one such cost-effective way of doing this. It has also been observed that acidic endoglucanase causes a better abrasion and less backstaining compared to neutral endoglucanase. For example, the cellulase given by
The latest trend of biostone washing is to utilize an enzyme mixture composed of amylase, cellulase, and laccase [12]. The sizing is the process by which denim material surface is covered by a compound like starch to provide rigidity and stiffness to raw denim and provide strength and friction resistance during handling. During washing, this surface layer of starch must be removed first to facilitate interaction between cellulases and cotton fibers. The amylase hydrolyzes starch from the fabric and causes desizing. Cellulase hydrolyzes small cellulose fibers, and laccase usually causes bleaching of the fabric. Laccases (EC 1.10.3.2) with intrinsic electron-donating tendency can decompose indigo in the solution as well as on the fabric creating bleaching effect on denim garments. The indigo dye is converted into isatin and anthranilic acid like chemical forms. This prevents backstaining of dye on the fabric surface. Eventually, this will give a complete faded look to the denim fabric. The purpose of using a mixture is to improve the efficiency of biostone washing process by allowing those three enzymes to work together in a sequential manner.
\nIn a recent study, it is reported that an alkali-stable cellulase in combination with xylanase from
These two processes are simply similar to each other. Cellulosic fibrous materials like cotton and linen are always loosing appearance because of fuzz formation on the fabric surface. Fuzz occurred due to short fibers protruding out from the surface of the fabric. Fuzz is sometimes loosely attached to the fabric forming a ball-like appearance which gives an unattractive look to the fabric. This is called pilling. The biopolishing process basically aims on removing microfibrils of cotton. It enhances fabric look, hand feel, and color by giving a smooth and a glossy appearance. This is also leading to improvement of color brightness, hydrophilicity, and moisture absorbance by the fabric [13]. The acidic cellulases produced by
The repeated washing of a cotton garment makes it fluffy and dull. This is due to partially removed microfibrils on the fabric surface. Biofinishing by cellulases can remove these fibrils and give back the smooth surface and original color to the fabric. This will also give a soft hand feel to the material, and also this is a good way of removing stains and dirt spots that are trapped within the cotton fiber network [14, 15].
\nThis is the process that removes noncellulosic material from the surface of the cotton. This is usually done with cellulase alone or in combination with other enzymes such as pectinase. Pectinase digests the pectin substance present among cellulose fibers. This helps to remove the intact connection between the cuticle and the main body of the cellulose fiber. This helps to degrade the primary cellulosic wall of the fiber. The ultimate result is the destruction of the cuticle [16]. This reaction increases the softness of the fabric.
\nThis is a kind of a biological mode of cleaning the fabric from the cellulosic or vegetative impurities with the help of enzymes. When a pure cotton or cotton blend fabric is prepared, some traces of unwanted cellulosic material still may remain in the fabric. They may result in imperfect finishing and lower quality of the fabric. The earliest methods of carbonization involved application of sulfuric acid. It was not only expensive but also corrosive, unsafe, and hazardous. Being a nonhazardous, non-corrosive, and eco-friendly method, enzymatic carbonization was a promising alternative. This method was perfect for removal of cellulosic impurities from the material because it was least affecting the color and the hand feel of the fabric. The removal of vegetative impurities from the surface of raw wool using cellulases is called wool scouring [17]. Cellulases can be used alone or in combination with other enzymes such as pectinases to increase the efficiency of this process. These methods are doing less damage to the fabric when compared to the treatment with sulfuric acid.
\nLyocell is the generic name for a biodegradable fabric that is made out of treated wood pulp. This material is used in everything from clothing to cars. This is obtained from wood pulp using a solvent-spinning method. The solvent system which is usually applied is an organic compound called N-methylmorpholine N-oxide. Some main characteristics of lyocell fibers are that they are soft, absorbent, and very strong when wet or dry and resistant to wrinkles. One chief defect of this material is fibrillation. This is the formation of small tangled fibrils on the surface of the fabric. Cellulases can be efficiently applied to remove these fibrils and give the fabric an increased softness and an improved appearance. This is also good for preventing fuzz and pill formation.
\nAlthough the applied enzymes are nontoxic and biodegradable in the above processes, the final effluent produced will show increased biological oxygen demand (BOD), chemical oxygen demand (COD), total dissolved solids (TDS), and total suspended solids (TSS) to a certain extent because the effluent may contain digested sugar and cellulosic forms. Direct release of this effluent to natural water bodies may cause water pollution. Alkalinity and pH fluctuations of the effluent will also result in polluted water which is not good for human and animal consumption. This may cause health issues like skin irritations in humans. On the other hand, the enzymatic treatments need an incubation period to facilitate the reaction between enzyme and fabric. As this is a fermentation reaction, this may release certain odors to the environment which may cause air pollution to a certain extent. Moreover, textile dyes are removed during textile processing. Most of the dyes are toxic and some are highly carcinogenic. Mixing this type of dyes with water is definitely causing adverse health effects in humans. Therefore, direct release of effluent without applying any pretreatments to neutralize these toxic compounds will breach the stability in ecosystems to which they are released. Therefore, establishment of pretreatment facilities and water quality testing procedures are essential for these enzymatic textile processing plants.
\nLyocell production has a different impact on the environment compared to the other textile polishing and finishing processes. The solvent which is used to manufacture this textile is N-methylmorpholine N-oxide. This is usually causing acute toxicity (oral, dermal, inhalation), skin irritation, serious eye damage and irritation, skin sensitization, and specific target organ toxicity. These are also hazardous to aquatic environments. These possible environmental impacts must be always addressed although application of enzymes in textile processing is eco-friendly.
\nThis is one of the largest industrial sectors in the world. According to the World Wildlife Fund (WWF), the pulp and paper industry, which includes products such as office and catalog paper, glossy paper, tissue, and paper-based packaging, uses over 40% of all industrial wood traded globally [18]. On the other hand, the latest paper industry statistics reveal China, the United States, and Japan as the three countries where the largest paper production occurs in the world. Half of the total paper manufacture of the world is done by these three countries. However, Germany and the United States are the world’s leading paper importers and exporters [19]. Moreover, the United States is reported to be the largest consumer of papers.
\nPapers and pulp are renewable resources. Therefore, recycling and reusing are two popular concepts related to this industry. Application of microbial cellulases is usually utilized for this purpose. The application of cellulases in this industry is broader. Starting from the 1980s up to now the possible applications are branching toward many areas. For instance, deinking, pulping, bioremediation of industry wastes, bleaching, and fiber enhancement can be taken.
\nThe drawbacks in mechanical pulping processes of woody raw materials such as refining and grinding resulted in pulps with higher amounts of fines, bulk, and stiffness. On the other hand, the process was high energy consuming which was not a profitable option for an industry. Meanwhile, biopulping using enzymes such as cellulases is an energy-saving way, and also it is eco-friendly [20]. The substantial energy saving is reported around 20%–40%. During the refining process, it generates small particles of the pulps. These particles reduce the drainage rate during the paper-making process. These particles can be readily degraded by cellulases in order to increase the drainage ability of the pulp. Mixtures of cellulases (endoglucanases I and II) and hemicellulases have also been used for bio-modification of coarse pulp material to improve fiber properties. It is strengthening the hand sheets. On the other hand, biological pulping has the potential to improve the quality of pulp and properties of the paper while reducing energy costs and environmental impact [21].
\nIn traditional deinking, large quantities of chemicals are used which make the method expensive and environmentally damaging and increase the release of contaminants [22]. The main advantage of bio-deinking is the ability of avoiding the alkali use during the process. This prevents yellowing of the paper. Cellulases alone, or used in combination with xylanase, are beneficial for deinking of different types of paper wastes. In most of the applications, partial hydrolysis of carbohydrate molecules releases ink from the fiber surface. This is done by a mixture of cellulases alone or in combination of cellulases and hemicellulases. The advantages associated with enzymatic deinking are clean look of paper, enhanced brightness, as well as environmental pollution reduction..
\nSuccessful application of cellulase and hemicellulase mixtures has been reported to modify properties of fibers. Usually in the paper industry, the making of paper is made easier by improving the beatability, runnability, and drainage of paper pulp during the process. Modifications of fiber properties are also achieved through treatment of paper by cellulase enzyme [20]. Not only that but also the enzymatic hydrolysis helps in characterization of fiber using various techniques such as scanning electron microscopy (SEM) and HPLC [23].
\nThe application of enzymes in manufacturing enzymatic washing agents or biological detergents dates back to the 1960s. Using enzymes in detergent formulae is a common practice today. In fact, according to market reports, by 2014, the detergent industry was the largest single market for enzymes at about 25–30% of total sales [22]. Another market research report published in 2017 on laundry detergent market stated that its global market size valued at 133.3 billion USD in 2016. The latest trend in the industry is to use alkaline enzymes in large amounts. For instance, protease, cellulase, α-amylase, lipase, and mannanase are broadly applied in heavy-duty laundry and automatic dishwashing detergents..
\nThe capability of enzymes to remove stains is the major focus of using them in manufacturing detergents. Cellulases are available in the market in different brands. For instance, Celluzyme® and Carezyme® are two main brands applied in detergent blends. These detergent blends are mainly applied in washing fabrics made of cotton and cotton blends. These detergents are making fiber modifications in the fabric in order to improve color brightness, softness, and particulate soil removal.
\nCellulases extracted from fungi like
The most recent innovation is to use combinations of enzymes in detergents. Cellulases are used in combination with other enzymes like proteases and lipases. The combination of enzymes is used to increase efficiency on stain cleaning and fabric care. For instance, SaniZyme® is a four-enzyme liquid detergent containing lipase, cellulase, amylase, and protease. This is a bacteriostatic enzymatic detergent for the removal of blood, protein, mucous, fats, lipids, and carbohydrates from all types of endoscopic equipment and surgical instruments. Another example is Getinge Clean MIS Detergent® which is also a formulation which includes protease, lipase, amylase, and cellulase enzymes, surfactants, sequestering agents, and corrosion inhibitors (typical pH in use dilution 8) which is specifically designed to clean complex, minimal invasive instrumentation.
\nThe application of cellulases in agriculture is usually reported in enhancement of crop growth and a control agent of plant diseases. For this purpose, combinations of cellulases, hemicellulases, and pectinases are broadly applied. Certain fungal cellulases are with the ability to degrade cell wall of plant pathogens. There are lots of details about application of bacteria such as plant growth-promoting rhizobacteria (PGPR) to improve plant performance. It is reported that these bacteria play a major role in reducing application of chemical fertilizers increasing plant development and also controlling potential plant pathogens and protecting plants from diseases. Moreover, many fungi including
According to available information, it is evident that cellulolytic microorganisms are participating in many processes, viz., rhizosphere soil decomposition, increasing the availability of nutrient for the plant, controlling plant pathogens, facilitating root colonization, and penetration of cereal crops improving yields and nutritional contents. However, as there are no solid evidence to prove the mechanisms behind these, this area needs further research. The studies should be performed in order to characterize and improve applications of microbial cellulases in this field.
\nDuring traditional agriculture practices, especially in countries like Sri Lanka, farmers used to add straw and
Medical pharmacology is currently a very active field of research that novel discoveries are coming into action. One such area is cellulases for development of medicine. By the way, humans are not cellulase producers, but the recent research on health and medicine reveals the benefits of consuming blends of enzymes including cellulase. As a result of global demand for enzyme blends, cellulase produced by the natural fermentation process of
In some records, the direct and indirect applications of cellulase in medicine have been mentioned apart from using it as consumable enzyme blends.
\nCellulase of fungal origin in combination with chitinases and lysozymes has a reported use in chitosan degradation. To obtain chitosan, a partial degradation of chitin must take place. As cellulose, chitin is a structural polysaccharide present in animals such as marine animals like shrimp and insect exoskeleton as well as participates in the formation of some parts of fungal cell walls. Chitin is a poly-ß-1,4-N-acetyl-D-glucosamine, conforming crystalline microfibrils. This polysaccharide provides structural integrity, stability, and protection to animals. Chitosan is the most important semicrystalline derivative form of chitin. This is obtained by partial deacetylation of chitin (around 50%, soluble in aqueous solution) under alkaline conditions or enzyme hydrolysis. Chitosan and its derivatives have many medical applications, viz., surgical sutures, bone rebuilding, production of artificial skin, anticoagulant, antibacterial agent, hemostatic dressings, anticancer and antidiabetic agents (in combination with metals), hypocholesterolemic effectors, elaboration of cosmetics, production of biopharmaceutics, and encapsulation of diverse materials [24, 25].
\nApart from these applications, a lot of reports have been published on several studies which discuss how cellulases hydrolyze chitosan and their potential biomedical effects. For instance, antitumor activity of cellulase-treated chitosan [26] and antimicrobial activity of low-molecular-weight chitosan obtained by
A bezoar is a mass found trapped in the gastrointestinal system. Phytobezoars, as its name suggests, are composed of indigestible plant material (e.g., cellulose). In other words, it is a gastric concretion formed by vegetable fibers, seeds and skins of fruits, and sometimes starch granules and fat globules trapped inside the gastrointestinal tract. This is a common problem frequently reported in patients with impaired digestion and decreased gastric motility. Although sometimes surgeries are needed to remove these stagnated substances, certain minor conditions can be treated by cellulases. The common application reported is fungal cellulases. As there are no much reports about application of bacterial cellulases, research can be conducted to find out the application of potential cellulolytic bacterial cellulases to treat this condition. However, what is most important is that any of these individual enzymes or enzyme cocktails should not adversely affect the healthy body cells.
\nAnother possible direct application of cellulase in medicine is degradation of cell walls of pathogenic organisms.
Pathogenic microorganisms usually form biofilms. A biofilm is an assemblage of microbial cells that is irreversibly associated (not removed by gentle rinsing) with a surface and is enclosed in an extracellular polymeric substance (EPS) matrix. Usually, pathogenic microorganisms form this type of assemblages. They may be found on a wide range of surfaces including living tissues and indwelling medical devices. Artificial hip prosthesis, central venous catheter, prosthetic heart valve, intrauterine device, and urinary catheter are some examples for indwelling medical devices that are commonly associated with biofilms. Most of the microorganisms produce extracellular polymeric substances composed of backbone structures that contain 1,3- or 1,4-β-linked hexose residues and tend to be more rigid, less deformable, and in certain cases poorly soluble. This is the exact linkages present in cellulose polymer. Therefore, cellulases can be further studied for their efficient application in removing this type of biofilms from medical devices.
\nFood is essential for all living organisms to obtain nutritional support for their growth and well-being. The huge demand for food has laid a path to a very complex, interconnected global business that supplies most of the food consumed by the world’s population.
\nFood biotechnology nowadays considers cellulases as an invaluable resource due to their increased applicability in a broad range of processes. Fruit and vegetable juice clarification, reducing the viscosity of nectars, concentrating purees, alteration of fruit sensory properties, carotenoid extraction, olive oil extraction, and the quality improvement of bakery products are among the various processes in food biotechnology that cellulase is exploited worldwide.
\nThe cloudiness which is usually present in fruit and vegetable juices is a result of floating polysaccharide materials such as cellulose, hemicellulose, lignin, pectin, starch, metals, proteins, and tannins. The presence of these materials in the juice makes it low quality and draws less consumer demand. “Rapidase pomaliq” is a commercially available enzyme preparation composed of cellulase, hemicellulases, and pectinases obtained from
Modification of sensory parameters of food is another important area where application of cellulases is highly recommended. The aroma properties, flavor, and texture of fruits are some sensory properties which play a crucial role in food biotechnology. The infusions of pectinases and cellulase enzymes have been found to be effective in altering the sensory properties of fruits and vegetables [29].
\nThese enzymes are also applied in degradation of grape fruit peels to release sugars. These sugars will be used in many industries including food production. Another important application of cellulase is extraction of phenolic compounds from grape pomace.
\nDuring extraction of olive oil, malaxing (mixing) is an indispensable step. This period allows the tiny oil droplets to attach with bigger ones and increase the oil yield which is coming from the olive paste. The use of cellulases alone or in combination with other hydrolytic enzymes like pectinases in this step has been found to have an enhancing effect on the extraction as well as the quality of olive oil. The enzymatic treatment of olive oil at the extraction stage causes significant enhancements in phenolic content and antioxidant activity of olive oil, thereby ultimately improving its quality.
\nThe enzyme cocktails of cellulases with other hydrolytic enzymes such as amylases, proteases, and xylanase result in increased loaf volume, improvement in bread quality, and production of softer crumb. Enzyme cocktail containing cellulases, hemicellulases, amylases, lipases, and phospholipases results in dough conditioning with improvement of flavor, prolonged shelf life, and increase in volume after baking [30].
\nAnother important application of cellulases is in pigment extraction from plants and plant products. Natural pigments such as carotenoids are nowadays earning a huge consumer demand as food colorants because of their natural origin, less toxicity, and availability of a wide range of colors. Brightly colored fruit peels such as orange, sweet potatoes, tomatoes, and carrot cell walls are rich in carotenoids. These are applicable as natural food colorants. The treatment of fruit peels with enzyme cocktails including cellulase leads to carotenoid extraction.
\nIn animal feed production, cellulases are applied to enhance digestibility of cereal-based food and to increase nutritive values for a higher quality of forages. There are reports about efficiently using
Furthermore, cellulases play an important part in increasing the rate and extent of fiber digestion. This can be used as a positive effect on natural gastrointestinal processes of the ruminants. The ultimate result will be the increased availability of absorbable nutrients by digestibility enhancement of fodder. The partial hydrolysis of lignocellulose materials also leads to better emulsification of food in the animal digestive tract resulting in ultimate improvement of nutrients availability.
\nEnergy is the life blood of the modern world. Fossil fuel, among all energy sources, holds the highest consumer demand. Since the industrial revolution, there is a drastic increase in fuel consumption. As fossil fuels are not renewable resources, the depletion of its natural deposits is inevitable. Therefore, we are in an era of limited and expensive energy. With the recent rise in fossil fuel prices, along with growing concern about its adverse effects on environment such as global warming caused by carbon dioxide emissions and subsequent climate change problems, biofuels have been gaining popularity. In our study area, we are focusing on bioethanol production using cellulolytic microorganisms as well as fermentative yeast using cellulose as the substrate.
\nBioethanol is a renewable form of energy. Especially, second-generation bioethanol production is an emerging trend because of the abundance of low-cost raw materials. The largest potential feedstock for this purpose is lignocellulosic biomass, which includes materials such as agricultural residues (corn stover, crop straws, and bagasse), herbaceous crops (alfalfa, switch grass), short-rotation woody crops, forestry residues, waste paper, and other wastes (municipal and industrial). Lignocellulose is the most abundant renewable biomass. The yield of lignocellulose can reach approximately 200 billon metric tons worldwide per year [34]. Bioethanol production from these feedstocks has certain advantages. It is an attractive method of disposing those lignocellulosic materials which is the nonedible part of plants. Less production of pollutants makes it environmentally friendly. Most importantly, bioethanol production using lignocellulosic biomass does not create any food insecurity because it does not utilize any food crops before harvesting. Finally, it is abundant all around the year as a raw material.
\nHowever, the major drawback of this production process is associated with the structure of lignocellulosic biomass. It mainly consists of lignin, cellulose, and hemicellulose which collectively form a very stable structure. In order to release fermentable sugars from this substrate, it needs to undergo a pretreatment process to break open the stable structure. One of the main focuses of this chapter is to discuss about the possible biological pretreatment of lignocellulosic biomass with special references to the current studies conducted by scientists. It also includes a description about our own attempts in this particular area of research.
\nBiomass contains about 40–50% of cellulose, a glucose polymer; 25–35% of hemicellulose, a sugar heteropolymer; 15–20% of lignin, a non-fermentable phenyl-propane unit; and lesser amounts of minerals, oils, soluble sugars, and other components. Biological pretreatment of lignocellulosic biomass uses the ligninolytic potential of certain microorganisms (fungi and bacteria and actinomycetes) to reduce the recalcitrant nature which is mainly caused by lignin component of the feedstock and enhance its digestibility by hydrolytic enzymes [35]. The breakdown of lignin barrier changes the structure of lignocellulose and enhances the access to the cellulose and hemicellulose carbohydrate components present.
\nBiological pretreatment seems to be a promising approach due to its low capital cost, low energy, and little dependence of chemicals, mild environmental conditions, eco-friendly nature, and the absence of inhibitor generation during the process which affects in bioethanol production. Moreover, this process does not release any toxic materials or any toxic effluents to the environment.
\nHowever, there are few limitations in this strategy. The main drawback against the industrial scale application is the prolonged incubation time consumed to achieve the efficient delignification [36]. This is because of low hydrolysis rate of the microorganisms. Another possible drawback that comes into mind is the possible consumption of carbohydrates as well as fermentable sugar formed by the same microorganisms used to pretreat the material. This is possible to take place because most of the lignolytic microorganisms are producing cellulolytic enzyme batteries as well. Then, the substrate left for fermentative organisms will be minimal which could consequently lead to lower bioethanol yields. Therefore, it is essential to have poor cellulolytic microorganisms for delignification process.
\nTo minimize this type of drawbacks in a biological way, it is possible to use cocultures or biofilms of efficient ligninolytic microorganisms. Introduction of fermentative yeast isolates into the same microbial coculture would also be a perfect approach. However, developing the most efficient microbial consortium is not that simple. Excessive laboratory-scale studies are required to understand the optimum physiological as well as biochemical parameter setup.
\nFungi are found to be more efficient in degrading lignocellulosic biomass. For instance, white-rot fungi, brown-rot fungi, and soft-rot fungi can be taken. The first two are basidiomycetes and soft-rot fungi are classified in ascomycete group.
\nAmong these efficient ligninolytic microorganisms that have been studied so far, white-rot basidiomycete fungi are found to be more versatile in the process. Most research has been concentrated on species such as
Recently, some bacterial laccases have also been characterized from
The biological pretreatment can be performed by growing the microorganism directly on the feedstock or using the enzyme extracts. Solid-state fermentation is the method of choice for biological delignification. Thus, from the reports available, it is evident that white-rot fungi and actinomycetes can be used to remove lignin from lignocellulosic substrates. However, further studies are required to shorten the incubation time and to optimize the delignification process.
\nThe importance of enhancing enzymatic hydrolysis has been increased because of the urgent need for efficient biological pretreatment processes. For this purpose it is essential to search for high enzyme-producing organisms from the natural environment. Selecting the most effective strain and its culture conditions can make the process more efficient. Another important aspect is to find unique microbial communities for biological pretreatment. These communities can be called consortia. The efficient biodegradation of lignocellulosic biomass could be achieved by the synergistic action of various bacteria and fungi in a microbial consortium. There are a number of advantages in using a microbial consortium for biological pretreatment. The increase of adaptability, improved productivity, improved efficiency of enzymatic saccharification, control of pH during sugar utilization, and increase in substrate utilization are some of them. With the development of biotechnology and molecular biology, the production of hyperlignolytic mutants by genetic modification of wild-type species is one approach that could be studied further. Furthermore, complete understanding of the theoretical basis behind the mechanisms of actions of these hydrolytic enzyme systems is very useful in the process of enhancing hydrolytic efficiency.
\nVarious process parameters affecting biological pretreatment like incubation temperature, incubation time, inoculums concentration, moisture, aeration, and conditions of pH have to be optimized. This must be done with well-planned laboratory-scale experiments. It is essential to pay attention to the microorganism used as well as the type of lignocellulosic material utilized because these parameters obviously change based on these two factors. Accessory enzymes are those enzymes which act on less abundant linkages found in plant cell walls. These include arabinases, lyases, pectinases, galactanases, and several types of esterases. Some studies have reported that addition of these accessory enzymes will improve hydrolysis efficiency.
\nRecently, several studies have been conducted in Sri Lanka on efficient lignocellulose-degrading microorganisms isolated from the natural environment. The effect of coculturing these fungal isolates for degradation of lignocellulosic material has also been reported. Coculturing of
The current progress in applications of cellulases is truly remarkable and attracting worldwide attention. It has already conquered the global market in an unbeatable way. Microbes are an attractive topic of interest for the production of cellulases due to their immense potential for cellulase production. However, it is apparent that more efficient species are still out there in the environment unnoticed by researchers. Further exploration and understanding of hidden mechanisms behind the activity of these enzymes are much more important. Microbial cellulases are preferred for their potential applications in a broad range of industries. Their ventures are expanding day by day. More and more researches are required to produce scientific knowledge to meet the growing demands for microbial cellulase. The advances in the emerging fields such as biotechnology, microbiology, and molecular biology will open up novel strategies to magnify the still-unlocked potentials of these enzymes. Eventually, it will be able to fine-tune the areas which still are dragging on the way to their utmost success.
\nSpecial thanks go to Research Assistant, Mr. K. Mohanan and Technical Officer Mrs. Kumuduni Karunarathna at Bioenergy and Soil Ecosystems Research Project, National Institute of Fundamental Studies, Kandy Sri Lanka and the National Research Council (Grant No: 12-021) for financial support.
\nNo conflicts of interest.
\nThe term “silicone” is not always used consistently, and should only be used to refer to polymeric materials, avoiding the relatively common confusion with the metallic element silicon (Si). Silicones are synthetic polymers containing Si─O─Si bonds and are used in many industries for their water repellency, ability to wet-out surfaces, high permeability to gases, stability in extreme temperatures, and resistance to thermal, radiation and chemical degradation. The variety of physical forms and physiochemical properties that silicones can display has led to their adoption in a diverse array of healthcare applications including medical devices and as active pharmaceutical ingredients (API) and excipients in medicines for over 60 years [1]. One class of silicone materials that has generated continued interest and research is silicone adhesives, specifically those self-adhering materials that do not require any activation immediately prior to use. Silicone adhesives are used as excipients in transdermal patches, and as skin contact adhesives in prosthetic and wound care device attachment. Recent investigations support the use of silicone based pressure sensitive adhesives for their skin-friendliness, but also to enhance the efficacy of the drug in transdermal drug delivery patch products. Recent silicone technologies like silicone based hybrid pressure sensitive adhesives promise potential performance advantages and improved drug delivery efficacy in transdermal drug delivery systems. Other silicone adhesive types are well known for their atraumatic removal from skin - an ability to remove cleanly from compromised skin without negatively impacting the wound healing process.
\nThis chapter will review silicone based adhesive technologies, applications and characterization, emphasizing those self-adhesive materials often used in skin contact applications. One type of silicone adhesive that is well established in the medical device industry but outside the scope of this work are room temperature vulcanizing (RTV) sealants. While these sealants are an interesting and useful class of materials, they will not be a focus of this chapter. Unlike the self-adhering adhesives discussed in this chapter, once fully crosslinked, the RTV sealants are non-tacky and rubbery and designed to form a permanent bond between substrates. These materials have a similar chemistry to silicone caulks commonly in the construction industry, and have found utility adhering materials to silicone elastomers, bonding parts of medical devices together, and acting as encapsulants and sealants in a variety of medical devices, including pacemakers [2].
\nWhile the term “silicone” persists in common vernacular, “polyorganosiloxane” is a more appropriate term, and has found acceptance in most scientific literature. Polyorganosiloxanes are organosilicon polymers, the most common of which are the trimethylsiloxy-terminated polydimethylsiloxanes (Figure 1) [3].
\nChemical structure of typical polydimethylsiloxanes.
The silicon in polyorganosiloxanes can be combined with one, two or three organic groups, commonly ─CH3, ─CH═CH2 or ─H, with the remaining valence(s) satisfied with oxygen [4]. Branched silicone structures are made possible by substitution of dimethyl siloxane units (i.e., (CH3)2SiO2/2) with those that contain additional Si─O connections (e.g., CH3SiO3/2 or SiO4/2) [4]. It is through the fact that different siloxane units can be combined with one another in the same molecule that the great variety of silicone compounds arises [3].
\nSilicones exhibit an inorganic backbone chain (Si─O)n and organic, (typically methyl) side groups [5]. It is this unusual combination and the resulting physiochemical properties that are responsible for many characteristics of the silicone adhesives. The silicon to oxygen bonds of the backbone are longer and more open than carbon to oxygen bonds permitting the characteristic flexibility of the siloxane chain. By way of comparison, the rotational energy around a ─CH2─CH2 bond is over four times greater than that of a typical (CH3)2Si─O bond. This flexibility is responsible for the characteristic low surface tension observed in silicones which allows them to quickly “wet out” onto surfaces including skin [5].
\nIn addition to increased flexibility, the silicon-oxygen bonds are also stronger than carbon-carbon bonds. The bond energy of a Si─O bond along the backbone of a silicone polymer is 452 kJ/mol while the typical C─C bond of the backbone of an organic polymer is only about 348 kJ/mol [5]. The inherently strong backbone of silicone polymers can help explain the acknowledged chemical stability silicone polymers possess toward a variety of degradation routes including moisture, UV, and a wide range of temperatures. This is equally important at very low and very high temperatures, where some types of silicones maintain their characteristic physical properties and utility from −100°C up to 260°C [6].
\nSilicones in general, are hydrophobic, (i.e., having little or no affinity for water), so one may anticipate silicones to be extremely lipophilic, given the common perspective equating hydrophobicity with lipophilicity (i.e., having a strong affinity with lipids). However, in the case of silicones, only relatively small silicone polymers are lipophilic. Polydimethylsiloxane (PDMS) polymers in excess of six to eight (CH3)2SiO units have little affinity with lipids while larger polymers are essentially lipophobic. These hydrophobic and lipophobic properties impact the ability to solubilize drugs, oils, botanicals and other traditional active ingredients into a silicone matrix [4]. The relatively poor miscibility of silicones with many compounds may be a key to the noted release efficiency of those same compounds from silicones.
\nSilicone pressure sensitive adhesives (PSA) are comprised of high molecular weight silanol-functional silicone polymers and silanol functional MQ siloxane resins. While a simple mixture of silicone polymer and resin can yield an adhesive with adequate peel adhesion and tack properties, sufficient cohesive strength is lacking. The silicone pressure sensitive adhesives most often used in medical applications are the product of a silanol condensation reaction between the polymer and resin components yielding a network structure, commonly referred to as standard pressure sensitive adhesives (Figure 2). These materials have suitable cohesive strength for medical device and transdermal drug delivery system applications, and upon removal from the skin the adhesive layer is removed intact. These adhesives are typically supplied in a volatile solvent which is removed during the coating process.
\nSchematic of the standard silicone PSA.
Silicone PSA have a long history of use in transdermal drug delivery systems but may also be used to attach prostheses and wound care devices. One recent innovative example of the utilization of silicone PSA in medical device attachment is the Embrace® MINIMIZE Silicone Scar Aid which consists of a silicone PSA coated onto silicone elastomer (rubber) sheeting. A unique applicator allows the dressing to be applied to relieve tension on healing skin to minimize scar formation [7, 8].
\nAnother application where silicone PSA have found wide acceptance is in the field of transdermal drug delivery. Second to the active pharmaceutical ingredient (API) or drug, the pressure-sensitive adhesive used in a transdermal drug delivery system can be viewed as the most critical component. Without proper and sustained adhesion to the skin, drug delivery from this dosage form does not occur.
\nMultiple transdermal drug delivery system (TDDS) designs are reported in the literature and are commercially available including reservoir, matrix, and drug-in-adhesive (DIA) systems; slight variants and combinations of each of these patch designs are also found. The functional requirements of the pressure sensitive adhesives in each patch design can vary with the design. (Table 1) [9, 10].
\nAdhesive functional requirement | \nPatch construction | \n||
---|---|---|---|
Matrix with rim adhesive | \nReservoir with rate controlling membrane/face adhesive | \nDrug-in-adhesive | \n|
Biocompatibility | \n+ | \n+ | \n+ | \n
Moisture resistance | \n+ | \n+ | \n+ | \n
Acceptable tack | \n+ | \n+ | \n+ | \n
Good adhesion | \n+ | \n+ | \n+ | \n
Good cohesive strength | \n+ | \n+ | \n+ | \n
Adherence to backing layer | \n+ | \n+ | \n+ | \n
Adherence to rate controlling membrane | \n\n | + | \n+ (in some cases) | \n
Compatible with drug and excipients | \n+ (in some cases) | \n+ | \n+ | \n
Permeable to drug and enhancers | \n\n | + | \n+ | \n
Cold flow resistance | \n+ (esthetic only) | \n++ | \n++ | \n
Stabilize drug and excipients | \n\n | \n | + | \n
Adhesive functional requirements for common transdermal patch designs.
Regardless of the patch design, basic requirements for the adhesive that is in direct contact with the skin include sufficient moisture resistance to stay adhered while perspiring and showering and biocompatibility (i.e., the adhesive must be non-irritating and non-sensitizing at a minimum). The adhesive must also have acceptable tack to adhere quickly on contact, good wetting behavior to achieve sufficient adhesion for the duration of wear (typically from 12 h to 7 days) and possess sufficient cohesive strength to enable removal without residual adhesive remaining on the skin. In most transdermal patch designs, the adhesive must also resist cold flow, or creep, the property of an adhesive to deform, especially at ambient temperature prior to use or at skin temperature when in use.
\nThe TDDS design with the most straightforward adhesive requirements is a matrix patch with a rim adhesive layer around the periphery of the patch. In this type of patch design, the adhesive functions are not significantly different from other device attachment applications as the adhesive must simply adhere the patch to the skin for the intended wear period. If the rim adhesive layer comes into contact with the drug loaded matrix layer, the adhesive must also be compatible with the matrix layer components. Resistance to cold flow for a rim adhesive is esthetically pleasing but does not result in unintended drug exposure or impact the drug contact surface area, so is not usually a mandatory function.
\nReservoir patch designs are typically characterized by a liquid reservoir compartment with solubilized API separated from the skin contact PSA by a semipermeable membrane. For a reservoir patch design with an adhesive layer across the face of the entire patch, the adhesive must adhere to the membrane and provide adequate adhesion to skin, as well as be compatible with the drug and allow diffusion of the drug and any penetration enhancers to the skin interface. The adhesive properties must be resilient to the drug and enhancer(s) reaching saturation in the adhesive layer.
\nIn a drug-in-adhesive (DIA) patch design, the adhesive plays an even greater role in the overall function of the patch. While this type of patch construction is clearly the easiest to manufacture, many formulation challenges exist, particularly with a monolithic (i.e., single layer) design. In addition to the requirements stated above, the adhesive matrix must also stabilize the API and excipients in either a dissolved or dispersed state, and allow controlled release of the drug and enhancers. Cold flow reduction is even more challenging in monolithic patch designs too, as they commonly require a greater adhesive coat weight than constructs that use face adhesive layers.
\nIt is unlikely that any single, off the shelf, adhesive system can meet the demands for all patch formulations and patch types. Silicone PSA, along with acrylic and polyisobutylene (PIB) PSA, are commonly used in transdermal patch applications. The end-use properties of silicone PSA (tack, adhesion, cohesive strength) can easily be modified or customized by varying the resin-to-polymer ratio, the degree of cross-linking and the residual silanol functionality during preparation. Silicone PSA are soluble in a variety of volatile polar and non-polar hydrocarbon solvents and additional customization may be achieved
Schematic of amine compatible silicone adhesives.
Silicone PSA is utilized in a variety of marketed TDDS either as the primary adhesive system or in combination with acrylic adhesives. Table 2 provides a list of commercial TDDS that utilize silicone PSA as a component of the patch construction as of the time of this publication, the respective actives, and other relevant information is also included. The table highlights the evolution of TDDS designs from the first silicone-containing reservoir patch in 1981 to recent approvals of more sophisticated microreservoir and multilayer designs that incorporate different adhesive types to achieve demanding dosage requirements.
\nDrug | \nPatch | \nMarketer | \nConstruction | \nSilicone PSA components | \n
---|---|---|---|---|
Nitroglycerin (1981) | \nTransderm-Nitro® | \nNovartis | \nReservoir | \nSilicone face adhesive layer | \n
Fentanyl (1990) | \nDuragesic® | \nJanssen Pharms | \nReservoir | \nSilicone face adhesive layer | \n
Estradiol (1996) | \nVivelle-Dot® | \nNovartis | \nMicroreservoir monolithic matrix | \nSilicone matrix adhesive continuous phase with acrylate polymer microreservoirs | \n
Nicotine (1997) | \nGeneric (OTC) | \nAveva | \nMultilayer matrix | \nSilicone matrix adhesive continuous phase with acrylate face adhesive | \n
Estradiol / Norethindrone Acetate (1998) | \nCombiPatch® | \nNoven | \nMicroreservoir monolithic matrix | \nSilicone matrix adhesive continuous phase with acrylate polymer microreservoirs | \n
Fentanyl (2005) | \nGeneric | \nMylan Technologies | \nDrug-in-adhesive monolitic | \nSilicone matrix adhesive continuous phase | \n
Fentanyl (2006) | \nGeneric | \nLavipharm Labs | \nMultilayer matrix w/ membrane | \nSilicone matrix adhesive, continuous phase and face adhesive layer | \n
Methylphenidate (2006) | \nDaytrana® | \nNoven | \nMicroreservoir monolithic matrix | \nSilicone matrix adhesive continuous phase with acrylate polymer microreservoirs | \n
Fentanyl (2007) | \nGeneric | \nActavis Labs | \nReservoir | \nSilicone face adhesive layer | \n
Fentanyl (2007) | \nGeneric | \nMayne Pharma | \nReservoir | \nSilicone face adhesive layer | \n
Rivastigmine (2007) | \nExcelon® Patch | \nNovartis | \nMultilayer matrix | \nSilicone face adhesive layer | \n
Rotigotine (2007) | \nNeupro® | \nUCB | \nMicroreservoir monolithic matrix | \nSilicone matrix adhesive continuous phase | \n
Capsaicin (2009) | \nQutenza® | \nAcorda | \nDrug-in-adhesive monolitic | \nSilicone matrix adhesive continuous phase | \n
Clonidine (2009) | \nGeneric | \nAveva | \nMultilayer matrix w/ membrane | \nSilicone matrix adhesive continuous phase with acrylate face adhesive | \n
Fentanyl (2011) | \nGeneric | \nMallinckrodt Inc | \nMultilayer matrix w/ membrane | \nSilicone matrix adhesive, continuous phase and face adhesive layer | \n
Estradiol (2012) | \nMinivelle® | \nNoven | \nMicroreservoir monolithic matrix | \nSilicone matrix adhesive continuous phase with acrylate polymer microreservoirs | \n
Estradiol (2014) | \nGeneric | \nMylan Technologies | \nMicroreservoir monolithic matrix | \nSilicone matrix adhesive continuous phase with acrylate polymer microreservoirs | \n
Commercial TDDS patches utilizing silicone PSA.
In recent years, the nomenclature for silicone PSA listed in the FDA Inactive Ingredient Database (IID) has been standardized to allow patch formulators to more easily identify prior use and maximum potency. Previously, reference to the use of silicone PSA in transdermal patches varied from a description of an adhesive laminate to numeric product codes. The preferred substance name for standard silicone adhesives is now dimethiconol/trimethylsiloxysilicate crosspolymer, and the preferred substance name for amine-compatible silicone adhesives is trimethylsilyl-treated dimethiconol/ trimethylsiloxysilicate crosspolymer. Reference is made to various types of adhesive with the addition of a nominal resin/polymer ratio [12].
\nSilicone and acrylic PSA chemistries as well as combinations of the two are commonly utilized in transdermal drug delivery [13]. The selection of the adhesive is typically drug and TDDS design specific and each adhesive type has its own advantages and disadvantages. Silicone adhesives may be more challenging during patch formulation due to the immiscibility with many drugs and common excipients in the silicone matrix; while acrylic adhesives are often easier to formulate due to the increased solubility of drugs and miscibility of excipients. However, higher drug utilization is often observed from TDDS that employ a silicone PSA over comparable patches that use an acrylic PSA [13, 14]. Yeoh [14] has provided a review of marketed fentanyl patches and has shown patches utilizing silicone adhesives have much greater fentanyl depletion during use and lower residual drug content after their intended use than comparable patches that use an acrylic adhesive. Minimizing the amount of residual drug in the patch at the end of the labeled use period, particularly with opiate drugs, is a focus of a recent FDA Guidance [15].
\nCombining silicone and acrylic pressure sensitive adhesives to form an immiscible polymer blend can provide benefits for transdermal drug delivery through selective modification of the solubility and/or diffusivity of the drug in the polymer blend matrix [16]. These micro-reservoir systems allow the drug to be solubilized in high concentrations in the discontinuous polyacrylate phase [17] and have been shown to be beneficial in decreasing patch size and required drug loading [18]. This technique has been successfully implemented in several commercial transdermal patches on the market including CombiPatch®, Daytrana® and Minivelle® (Noven Pharmaceuticals) as well as Vivelle Dot® (Novartis Pharmaceuticals) [16] A review of label claims for two patches that provide a 0.5 mg/day dose of estradiol reveals that a 5 cm2 Vivelle Dot® patch, which employs the Dot Matrix® technology, can deliver 22.4% of the drug, whereas the 12.5 cm2 Climera® with an acrylic PSA construction only delivers 9.0% of the drug [19]. These immiscible blends do have a major limitation in that they will exhibit macro phase separation in the coating mass if mixing is discontinued which may be exacerbated upon addition of other formulation ingredients such as penetration enhancers [20]. One potential means to prevent macro phase separation of the two immiscible adhesives is to covalently link the two polymer chemistries together, creating a silicone-acrylate hybrid material.
\nHybrid adhesives, in which silicone and acrylic chemistries are combined, have been described following different routes [21, 22]. One approach is the reaction product of a (meth)acrylate-functional silicone PSA and ethylenically unsaturated monomers, [21] whereas a second route toward a hybrid adhesive describes an alkoxysilyl-functional acrylic prepolymer that is further condensed or “bodied” with silicone PSA precursors (i.e., OH-functional silicate resin and OH-terminated PDMS) in the presence of a condensation catalyst [22]. These hybrid adhesives, although produced
Optical micrograph (100X magnification) of (A) 50:50 blend of silicone PSA and non-functional acrylic PSA and (B) silicone-acrylate hybrid adhesive (50% acrylate) [
Drug delivery using silicone-acrylate hybrid adhesives (SilAc I and SilAc II) differing in the ratio of high and low Tg acrylic monomers has been reported, and delivery of estradiol (Figure 5A), clonidine (Figure 5B), and ketoprofen was demonstrated across human cadaver epidermis from these matrices. The authors also noted that the use of silicone-acrylate hybrid PSA, singularly or as blends with silicone PSA resulted in a more desirable wet blend compatibility/stability than those obtained with blends [23].
\nDrug flux from silicone-acrylate hybrid PSA based patches; (A) estradiol 1.5 wt%; (B) clonidine at 1, 1.5 and 2.5 wt%; [
Due to the inherent immiscibility of silicone and acrylate polymers, the hybrid adhesives contain micro-domains which can be observed using transmission electron microscopy (TEM) as presented in Figure 6. Further analysis of the phase behavior reveals the ability to selectively control the domain arrangement (i.e., silicone-in-acrylate or acrylate-in-silicone) of these materials by the choice of casting solvent, with the phase having the highest affinity with the casting solvent remaining external, (i.e., heptane casting solvent exhibiting a silicone continuous phase and polyacrylate discontinuous phase (Figure 6A) or
Transmission electron micrograph of silicone-acrylate hybrid adhesive films, silicone phase appears dark due to the electron density (A) cast from heptane and (B) cast from ethyl acetate.
The selective control of the phase arrangement provides potential options for tuning both the adhesive properties as well as tailored drug release profiles as illustrated in Figure 7.
\nIllustration of silicone-acrylate hybrid adhesive microstructure and potential impact on drug solubility and release.
The impact of casting solvent and silicone content on the material properties has been conducted using a dynamic rheometer (Figure 8A). Blends of silicone PSA and silicone-acrylate hybrid PSA (nominally 50% silicone) were prepared in either heptane or ethyl acetate to yield a range of materials. For materials delivered from ethyl acetate, between 76% and 78% silicone, a precipitous change in tan delta is observed followed by incremental decrease as the silicone content rises. Tan delta is a rheological property that approximates the internal friction of a material. When tan delta is greater than one, a material is more viscous than elastic, and when it is less than one it is more elastic than viscous. TEM analysis suggests this is the result of phase inversion when the silicone becomes the external phase. This change is not observed for materials delivered from heptane as the silicone remains the external phase over the entire range. Films containing either 1.0 wt% estradiol (Figure 8B), 2.5 wt% ibuprofen (Figure 8C), or 2.5 wt% lidocaine (Figure 8D) were prepared using blends of hybrid PSA with silicone PSA to investigate the impact of phase arrangement on the release behavior. All three API demonstrate a change in drug release characteristics between 75 and 80% silicone content, which is where rheology suggests the phase inversion occurs [24].
\nRheology and drug release as a function of silicone content and dispersion solvent; (A) tan delta of the adhesive matrix; (B) estradiol (E2) 6 h cumulative release; (C) ibuprofen (IBU) 1 h cumulative release; (D) lidocaine (Lido) 1 h cumulative release [
The characterization of PSA materials is a critical part of innovation development and production quality control. Historically, tape properties such as peel adhesion, shear and tack have been used to characterize the performance of pressure sensitive adhesives targeted for transdermal applications. However, these tests often have high variability resulting in wide specification limits and poor correlation of test data with adhesive performance in real life applications [25]. Furthermore, tape property tests can be substrate dependent. That is to say, they are influenced by the substrate on which the PSA is coated and also by the substrate on which the adhesive performance is measured. Despite the drawbacks of tape property testing, they are still commonplace and so, warrant some discussion.
\nPeel tests are well described in the literature and are common to the majority of adhesives. The peel test typically occurs at 90o or 180° and the force to remove the adhesive from a substrate (e.g., stainless steel in many cases) is measured. In the case of silicone PSA, the typical adhesive thickness tested is relatively thin, commonly between two and five mil (approximately 51–127 micron). A distinction between peel adhesion and tack of an adhesive is often made. From an analytical test perspective, the distinction between peel adhesion and tack measurements is the time allowed for the adhesive to bond with the substrate. When measuring tack, the measurement is taken almost instantaneously after the adhesive comes in contact with the test substrate, whereas peel adhesion is measured after the adhesive is left in contact with the substrate for a longer time period. The time between application and testing allows the adhesive to wet out on the surface and the adhesion to build.
\nShear testing may have greater relevance to skin contact adhesive applications than the aforementioned peel adhesion and tack tests. Since PSA are condensed materials that have the ability to flow, the extent of cold flow must be characterized to fully understand and anticipate the surface area of adhesive in contact with skin, which can impact the amount of drug delivered from a transdermal patch. Shear tests of fully formulated adhesive matrices may be even more relevant to the performance of the final TDDS. If the skin/adhesive interface changes over time, the transdermal drug diffusion will also change. Typically, a shear test is the measurement of the time for the adhesive to detach from a surface (e.g., stainless steel) under a constant weight.
\nThe advantages of tape property test methodology include ease of set up, reproducibility and a straightforward interpretation of data. However, drawbacks including the considerable influence adhesive coating thickness has on the test, the influence of the substrate on which the adhesive is coated, and the surface on which the test is conducted must also be rationalized. To minimize these influences, there must be accurate control of adhesive thickness and standardization of substrates and test surfaces.
\nAlthough tape property testing may qualitatively predict how quickly a system may bond to a substrate, the extent to which the adhesive resists cold flow, and how much force may be needed to remove it, and perhaps most importantly, the wear performance of the system may not be adequately addressed using classical characterization techniques. In order to better understand and predict the wear performance of transdermal systems, rheology is often used to understand the adhesive bulk viscoelastic behavior. [26] Rheological characterization allows the analyst to overcome the inherent uncertainty linked to peel, tack and shear tests by minimizing the influence of sample preparation and substrate variability on adhesive characterization results. Rheology is a technique to characterize viscoelastic properties of polymers and also predict wear performance of pressure sensitive adhesives. As shown below in Figure 9, a typical rheological curve can be correlated to tape properties [27, 28, 29, 30].
\nA schematic representation of the link between the rheological profile and the final pressure sensitive (PSA) wear performance [
Data have shown that for viscoelastic materials, such as silicone pressure sensitive adhesives, frequency sweep curves are sensitive to structural differences (e.g., crosslink density) and formulation changes (e.g., resin-to-polymer ratio). This sensitivity provides a means to identify, characterize and predict adhesive wear performance [26].
\nStorage modulus (G′) is an indicator of how elastic the adhesive is and how much energy is stored during deformation, while the loss modulus (G″) indicates the viscous component of the PSA and how much energy is lost as heat, while complex viscosity (η*) is an indicator of the adhesive bulk viscosity and can be related to the cold flow [25]. Bonding of a transdermal system occurs at a low deformation rate, and is dependent on the wetting behavior of the adhesive when it comes into contact with skin [26]. Rheologically, the storage modulus, G′, values at low frequency may be used for predicting wetting and creep (cold flow) resistance. Optimum wetting occurs when the adhesive modulus is low. Subsequently, debonding of a transdermal system occurs at high deformation rates [26].
\nRheologically, the storage modulus, G′, and loss modulus, G″, at high frequency may be related to the peel adhesion and quick stick (i.e., tack) properties of an adhesive and the subsequent TDDS [31, 32]. For bonding, the viscous contribution should be higher than the elastic contribution to the PSA viscoelastic profile. In rheological terms, this means that at low frequencies, G′ < G″ and the opposite for the debonding step, represented at high frequencies where G′ should be equal to or higher than G″. Based on this interpretation, the rheological traces in Figure 10 suggest that the increase of resin content should lead to reduced cold flow (i.e., an increase of the complex viscosity with resin content) and an increase of the adhesion strength (i.e., increase of both G′ and G″ with resin content). Dynamic frequency sweeps (0.01–100 rad/s) were conducted on dried adhesive solids using a TA ARES-G2 rheometer. The adhesives with high and medium resin content were tested using 8 mm parallel plates, at 0.35% and 0.5% strain respectively. The adhesive with low resin content was tested using 25 mm plates, at 0.5% strain. All samples were tested at 30°C with a 1.5 mm gap.
\nTypical frequency sweeps of silicone PSA at three common resin contents.
In the early 1990s, E.P. Chang developed a theory to interpret rheological data of pressure sensitive adhesives and establish criteria for PSA classification when used in conjunction with the Dahlquist’s criteria [33]. This theory is now well known as “Chang viscoelastic window.” As depicted in Figure 11, a G′ vs. G″ graph, is divided into four quadrants with a central axis. The location of the analyzed PSA within this graph allows a straightforward extrapolation from rheological properties to real-world adhesion performance. For example, the top right hand quadrant corresponds to high modulus and high dissipation. Therefore, materials in this quadrant with characteristically high G′ modulus compensated by the high G″ are anticipated to be adhesive materials with high adhesion but low tack and high shear resistance. Conversely, the bottom left quadrant corresponds to low modulus and low dissipation; these materials, are anticipated to exhibit low peel values because of the comparatively low debonding cohesive strength and low dissipation.
\nChang viscoelastic window concept adapted for low resin content silicone pressure sensitive adhesive (PSA) with differing amounts of isopropyl myristate (IPM) [
Changes in the Chang viscoelastic window, of a typical low resin content silicone PSA can be observed as differing amounts of a commonly used permeation enhancer, isopropyl myristate (IPM), are added (Figure 11) [34]. The Chang viscoelastic window of the neat adhesive moves from the upper right quadrant to the lower left quadrant as more IPM is added. The lowermost edge of the window which is linked to bonding of the adhesive is far below Dahlquist’s criteria, so the adhesive would be expected to have reasonable tack. There is a significant shift in the position of the upper right corner as IPM content increases which is linked to debonding (peel) efficiency suggesting that an increase of IPM content decreases peel efficiency [34]. Finally, the window size increase indicates a decrease of the PSA shear strength likely due to better solvent compatibility in the PSA. These data coincide with observed changes in adhesive properties as plasticizing agents like IPM are added and support the further use of rheological measurements to characterize changes in wear properties.
\nSilicones have more than 30 year history of safety and efficacy in advanced wound care applications. Much of the success of silicones in wound care is due to an adhesive technology referred to in the literature by many names including soft skin adhesives (SSA), tacky gels, silicone gels and silicone tacky gels among others [35]. The technology was introduced to the wound care market by Dow Corning Corporation in the 1990s and similar materials are offered today by many silicone suppliers under a variety of brand names [36, 37, 38]. In a segment that was historically controlled primarily by acrylic adhesives, the tacky gel technology concept was disruptive by securing wound dressings while providing gentle adhesion upon removal. SSAs have become the material of choice in many advanced wound care applications, due to their reliable adhesiveness, while being easier to remove and causing less pain than many other adhesive technologies of the day.
\nSSAs are based on a polydimethylsiloxane network which supports the critical adhesive attributes required for securing the device in place and removing it without leaving residue or damaging the skin. Unlike silicone PSAs that build their adhesiveness on a viscous phase bodied with a silicate resin, SSAs are based on the silicone elastomer technology modified to deliver the relevant visco-elastic profile. They also differ from analogous silicone elastomers (e.g., liquid silicone rubber (LSR) technology) by the absence of reinforcing silica filler. As a result, they have a similar consistency to gels, but SSAs are not a typical polymeric gel because they are not based on an insoluble polymer network swollen with fluids. The visco-elastic behavior of SSA also differs from silicone PSA, despite their low consistency and a high degree of compressibility, SSAs show resilience and quick recovery under cyclic deformation [35].
\nThe pressure sensitive adhesive property of SSAs are based on the capacity of the elastomer surface to quickly wet the skin and conform to skin irregularities without an additional compression step as required for a silicone PSA [35]. Thanks to the low intensity of the viscous component of the SSA rheological profile, the adhesive does not flow significantly, and very little dissipation of the energy occurs when deformation pressure is applied to the SSA. As a result, SSA debonding happens at low peel force, without skin stripping and painful skin pulling when the adhesive device is removed. Being elastomeric by nature, SSAs have a low viscous component that limits their flow and consequently the ability to pick up materials on or from the surface of the skin [35]. Therefore, unlike silicone PSA, the adhesive surface of SSAs remain relatively clean upon removal from the skin, allowing for removal and easy reapplication of the dressing or device to the skin, making wound dressing repositioning possible.
\nThe elastomeric structure of SSAs is obtained by cross-linking a network of polydimethylsiloxane (PDMS). The reaction is based on an addition reaction (hydrosilylation) between vinyl functional PDMS (polymer) and hydrogen functional siloxanes (cross-linker) as shown in Figure 12. The cure reaction is catalyzed by a platinum complex, which can occur at room temperature or be accelerated at elevated temperature (80–145°C), without the formation of reaction by-products [35]. As thermoset materials, SSAs have a low susceptibility to cold flow and plasticizing effects.
\nTypical hydrosilylation reaction schematic.
The SSA technology has been extensively used in scar treatment and advanced wound management, demonstrating safety and efficacy recognized by wound care professionals [35]. The use of SSA may be recommended when designing medical adhesive devices, tapes, bandages, drapes, and wound dressings and have been noted for the many benefits including high tack for quick bonding to skin, reliable adhesiveness and cohesiveness, gentle adhesion to fragile and compromised skin, no skin stripping and pain-free removal of the device, as well as permeability to moisture and gases (e.g., CO2, O2) [35].
\nSSAs are supplied as two-part systems with the catalyst in one part and the cross-linker in the other. The materials are characteristically transparent before and after curing into a solid matrix. They are typically processed by mixing the two parts and coating the mixture directly onto the final substrate (i.e., backing film), understanding that this film must be impermeable enough to prevent the uncured liquid SSA from wicking through. The typical coat weight for SSA can vary widely depending on the desired final properties, but often range between 150 and 250 g/m2. The curing phase is typically completed at elevated temperature adjusted according to the temperature sensitivity of the substrate. After cooling, the adhesive surface is protected by a release liner which is peeled off when the end user applies the adhesive to skin.
\nSubstrate selection is important when designing an adhesive device based on SSA, as the nature of the substrate can significantly impact the coating and cure conditions during the manufacturing phase. The anchorage of the adhesive to the substrate and the cohesion of the adhesive after cure, as well as the ultimate wear behavior of the device when applied to the body can all be impacted by the substrate selection.
\nThe choice of release liner is also a critical factor as it can affect the device stability, making it unusable if this protective film cannot be easily removed from the adhesive prior to use. Traditional silicone release liners that are used ubiquitously with acrylic adhesives cannot be used with SSA as the silicone release liner chemistry is similar enough to SSA that they are highly likely to interact and experience an irreversible lock-up effect upon storage. However, uncoated polyethylene films, especially LDPE (low density polyethylene) grade, can provide an acceptably low and reasonably consistent release force from the SSA [39].
\nNew SSA technology are being developed that can achieve higher adhesion and longer wear times as well as improved drug compatibility to address emerging medical system market trends including wearable devices and topical drug delivery patches [35]. The use of SSA technology to formulate drug delivery matrices enables drug delivery system designs which address the needs for secure and gentle fixation to fragile, sensitive or compromised skin conditions common in dermatology, wound care, pediatrics and gerontology. Several studies were conducted to evaluate the compatibility of various drugs and their release from SSA matrices. A variety of API have been studied including those indicated for pain relief and local anesthesia, antibiotics, and dermatological actives [39]. Wound care products that utilize silicone tacky gels as the skin contact adhesive and are loaded with chlorhexidine gluconate and other antimicrobial agents have also been investigated [40]. This may signal further interest in the utilization of SSA in even more advanced active-loaded therapies in addition to the traditional wound therapies where it has been used historically.
\nMany of the analytical techniques used to characterize silicone PSA have been modified to characterize the SSA materials, although shear tests are less emphasized for SSA due to the characteristically low cohesion of the SSA. In addition to adhesive peel measurements, the measurement of the softness of the SSA by penetration test is often performed. Over a broad range, the penetration measurement shows correlation to adhesion performance values within a formulation type and is linked to the adhesive network chemistry; therefore, it is often used as a quality control measurement.
\nPeel tests are commonly used in the adhesive industry, because for many applications these relatively easy to perform tests fit well with the final application of the adhesive. The substrates upon which most adhesives are tested to evaluate adhesive strength (e.g., stainless steel) often are not predictive of the relative strength SSA will exhibit in practice on skin. Therefore, some users have resorted to using substrates that have a surface energy more similar to that of skin as the test substrate for SSA. The number and diverse composition of substrates including plastic films, paper and even artificial skin materials, make standardization across the industry difficult, and comparison between users problematic. Testing is conducted similarly to that described for PSA, with the SSA typically being cast and cured at a consistent thickness directly onto a film. This substrate may influence the peel adhesion result due to its intrinsic elasticity and also potentially through interactions with the SSA. The gel on the backing substrate is then applied on a test substrate, taking care to apply the adhesive with a constant force. After a designated equilibration time, the adhesive is peeled from the substrate, typically at a 180° angle, and the force required to remove it is measured.
\nWhile this method provides relative adhesion strength, allowing comparison of adhesive values, the results may be significantly influenced by the backing substrate, as well as the test substrate used, so the results do not necessarily simulate the application of the adhesive to skin.
\nRheological measurements have been developed and used for decades to characterize silicone PSA and provide more realistic predictions of real-world adhesive performance than classical peel tests are capable of providing. Recently, similar rheological measurements have been applied to characterize the intrinsic properties of the SSA and offer a characterization method more capable of harmonization across the industry. The SSA rheological characterization is performed on free standing gels and is able to characterize the adhesive properties without the influence of backing or test substrates unlike the aforementioned adhesion tests. SSAs may be characterized in dynamic oscillation modes, using strain and frequency sweeps to measure the viscoelastic characteristics (e.g., storage modulus, G′ and loss modulus, G″). Different SSA, which exhibit significant differences with respect to adhesion can also be discriminated using rheological analysis. Identifying the true viscoelastic properties of the adhesives is critical to understand the adhesion performance of such products. Using the data generated from the rheometer, it is possible to correlate viscoelastic properties to adhesion, and to better understand structure-property relationships.
\nTo understand the rheological characteristics of this material one must identify the linear viscoelastic (LVE) zone by submitting the sample to an oscillatory strain sweep analysis. In the LVE zone, the elastic modulus (G′) and the loss modulus (G″) are independent of the shear strain, indicating that within this strain zone, the response of the material does not depend on the strain applied, and there are no modifications of the material structure. In the LVE zone identification test, the strain is the only parameter which varies, all other parameters, (e.g., temperature and oscillation frequency) are fixed. The LVE graph for the SSA exhibits a large linear viscoelastic zone from 0.5 to 30% logarithmic strain, providing some flexibility to set the strain when performing the frequency sweep at a fixed strain is the next step of the measurement process. Knowing the LVE zone of the material allows one to carry out the second phase of the rheological evaluation, the oscillatory frequency sweep test. Previously unreported data is shown to elucidate this concept in Figure 13. Samples were prepared by weighing equal amounts (±2%) of the two parts of the SSA and mixed to ensure homogeneity and then were degassed in a vacuum chamber. The mixed, uncured SSA was coated onto a polytetrafluoroethylene (PTFE) film at a thickness of 0.9 mm, and placed in a forced air oven at a temperature of 130°C for 4 min to cure the SSA. The cured laminate was removed from the oven and allowed to cool to ambient temperature. A second PTFE film was applied using a 6.8 kg (15 lb.) rubber coated roller to ensure complete and consistent contact between SSA and PTFE. The film was allowed to rest for 24 h after which a disc was cut from the SSA laminate using a 24 mm stainless steel punch. Dynamic frequency sweeps (1–100 rad/s) were conducted on SSA with a TA ARES-G2 rheometer at 32°C using 25 mm stainless steel parallel plates and a gap of 0.5 mm with a 10% strain (in the linear viscoelastic region). Data collection was set for 5 pts./decade.
\nFrequency sweep of three typical SSA.
The frequency sweep test is the most suitable rheological test to assess SSA adhesive properties in the final application. The viscoelastic behavior at low frequencies is related to the bonding step which occurs at low deformation rates and is linked to the SSA ability to wet the surface. Alternatively, the viscoelastic behavior at high frequencies is related to debonding (peel) which occurs at high deformation rates and is linked to the elasticity and energy dissipation during the removal. SSAs with varying adhesive levels can be effectively discriminated based on their rheological profiles. The rheological characterization agrees with the results experienced by skin adhesion, where adhesives with higher G′ and G″ provide higher skin adhesion.
\nThis rheology methodology should be an effective tool and a suitable starting point to understand the structure-property relationships of the SSA technology. It should also provide a means to separate the innate adhesive performance from the influences of substrates. Understanding the relationships between the SSA chemistry, adhesion and rheological profiles will provide key and essential information on structure-property relationships to push the boundaries of SSA even further.
\nSilicone adhesives have been safely and effectively used in a variety of medical applications and are notably present in drug delivery and wound care applications because of the unique benefits and properties provided. Continued investigation has resulted in recent, innovative product developments using established silicone adhesive technologies including innovative TDDS designs, wound care devices that prevent scar formation and those that are loaded with antimicrobial actives. Adhesive chemistry research has resulted in novel chemistries that combine seemingly incompatible acrylate and silicone adhesive technologies, whereas advances in measurement techniques have brought about clearer understanding of adhesive structure property relationships, avoiding many pitfalls experienced by previous researchers. Despite being used for several decades, the number and variety of recent developments suggest that identifying new medical applications of silicone adhesives remains relevant and the extent to which it may be used has not yet been tapped.
\nThe authors wish to acknowledge the following without whom this work would not have been possible; Chana Evans, Dave Gantner, Roger Gibas, Tim Mitchell, and Audrey Wipret. We would also like to acknowledge the work of Dr. Meng Gu and the microscopy team at The Dow Chemical Analytical Department for their assistance and collaboration.
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He received his Ph.D. in Molecular Biology with his thesis “Genetic variability of the tick-borne encephalitis virus in natural foci of Novosibirsk city and its suburbs.” His primary field is molecular virology with research emphasis on vector-borne viruses, especially tick-borne encephalitis virus, Kemerovo virus and Omsk hemorrhagic fever virus, rabies virus, molecular genetics, biology, and epidemiology of virus pathogens.",institutionString:"Russian Academy of Sciences",institution:{name:"Russian Academy of Sciences",country:{name:"Russia"}}},{id:"310962",title:"Dr.",name:"Amlan",middleName:"Kumar",surname:"Patra",slug:"amlan-patra",fullName:"Amlan Patra",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/310962/images/system/310962.jpg",biography:"Amlan K. Patra, FRSB, obtained a Ph.D. in Animal Nutrition from Indian Veterinary Research Institute, India, in 2002. He is currently an associate professor at West Bengal University of Animal and Fishery Sciences. He has more than twenty years of research and teaching experience. He held previous positions at the American Institute for Goat Research, The Ohio State University, Columbus, USA, and Free University of Berlin, Germany. His research focuses on animal nutrition, particularly ruminants and poultry nutrition, gastrointestinal electrophysiology, meta-analysis and modeling in nutrition, and livestock–environment interaction. He has authored around 175 articles in journals, book chapters, and proceedings. Dr. Patra serves on the editorial boards of several reputed journals.",institutionString:null,institution:{name:"West Bengal University of Animal and Fishery Sciences",country:{name:"India"}}},{id:"53998",title:"Prof.",name:"László",middleName:null,surname:"Babinszky",slug:"laszlo-babinszky",fullName:"László Babinszky",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/53998/images/system/53998.png",biography:"László Babinszky is Professor Emeritus, Department of Animal Nutrition Physiology, University of Debrecen, Hungary. He has also worked in the Department of Animal Nutrition, University of Wageningen, Netherlands; the Institute for Livestock Feeding and Nutrition (IVVO), Lelystad, Netherlands; the Agricultural University of Vienna (BOKU); the Institute for Animal Breeding and Nutrition, Austria; and the Oscar Kellner Research Institute for Animal Nutrition, Rostock, Germany. In 1992, Dr. Babinszky obtained a Ph.D. in Animal Nutrition from the University of Wageningen. His main research areas are swine and poultry nutrition. He has authored more than 300 publications (papers, book chapters) and edited four books and fourteen international conference proceedings.",institutionString:"University of Debrecen",institution:{name:"University of Debrecen",country:{name:"Hungary"}}},{id:"201830",title:"Dr.",name:"Fernando",middleName:"Sanchez",surname:"Davila",slug:"fernando-davila",fullName:"Fernando Davila",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/201830/images/5017_n.jpg",biography:"I am a professor at UANL since 1988. My research lines are the development of reproductive techniques in small ruminants. We also conducted research on sexual and social behavior in males.\nI am Mexican and study my professional career as an engineer in agriculture and animal science at UANL. Then take a masters degree in science in Germany (Animal breeding). Take a doctorate in animal science at the UANL.",institutionString:null,institution:{name:"Universidad Autónoma de Nuevo León",country:{name:"Mexico"}}},{id:"309250",title:"Dr.",name:"Miguel",middleName:null,surname:"Quaresma",slug:"miguel-quaresma",fullName:"Miguel Quaresma",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/309250/images/9059_n.jpg",biography:"Miguel Nuno Pinheiro Quaresma was born on May 26, 1974 in Dili, Timor Island. He is married with two children: a boy and a girl, and he is a resident in Vila Real, Portugal. He graduated in Veterinary Medicine in August 1998 and obtained his Ph.D. degree in Veterinary Sciences -Clinical Area in February 2015, both from the University of Trás-os-Montes e Alto Douro. He is currently enrolled in the Alternative Residency of the European College of Animal Reproduction. He works as a Senior Clinician at the Veterinary Teaching Hospital of UTAD (HVUTAD) with a role in clinical activity in the area of livestock and equine species as well as to support teaching and research in related areas. He teaches as an Invited Professor in Reproduction Medicine I and II of the Master\\'s in Veterinary Medicine degree at UTAD. Currently, he holds the position of Chairman of the Portuguese Buiatrics Association. He is a member of the Consultive Group on Production Animals of the OMV. He has 19 publications in indexed international journals (ISIS), as well as over 60 publications and oral presentations in both Portuguese and international journals and congresses.",institutionString:"University of Trás-os-Montes and Alto Douro",institution:{name:"University of Trás-os-Montes and Alto Douro",country:{name:"Portugal"}}},{id:"38652",title:"Prof.",name:"Rita",middleName:null,surname:"Payan-Carreira",slug:"rita-payan-carreira",fullName:"Rita Payan-Carreira",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRiFPQA0/Profile_Picture_1614601496313",biography:"Rita Payan Carreira earned her Veterinary Degree from the Faculty of Veterinary Medicine in Lisbon, Portugal, in 1985. She obtained her Ph.D. in Veterinary Sciences from the University of Trás-os-Montes e Alto Douro, Portugal. After almost 32 years of teaching at the University of Trás-os-Montes and Alto Douro, she recently moved to the University of Évora, Department of Veterinary Medicine, where she teaches in the field of Animal Reproduction and Clinics. Her primary research areas include the molecular markers of the endometrial cycle and the embryo–maternal interaction, including oxidative stress and the reproductive physiology and disorders of sexual development, besides the molecular determinants of male and female fertility. She often supervises students preparing their master's or doctoral theses. She is also a frequent referee for various journals.",institutionString:null,institution:{name:"University of Évora",country:{name:"Portugal"}}},{id:"283019",title:"Dr.",name:"Oudessa",middleName:null,surname:"Kerro Dego",slug:"oudessa-kerro-dego",fullName:"Oudessa Kerro Dego",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/283019/images/system/283019.png",biography:"Dr. Kerro Dego is a veterinary microbiologist with training in veterinary medicine, microbiology, and anatomic pathology. Dr. Kerro Dego is an assistant professor of dairy health in the department of animal science, the University of Tennessee, Institute of Agriculture, Knoxville, Tennessee. He received his D.V.M. (1997), M.S. (2002), and Ph.D. (2008) degrees in Veterinary Medicine, Animal Pathology and Veterinary Microbiology from College of Veterinary Medicine, Addis Ababa University, Ethiopia; College of Veterinary Medicine, Utrecht University, the Netherlands and Western College of Veterinary Medicine, University of Saskatchewan, Canada respectively. He did his Postdoctoral training in microbial pathogenesis (2009 - 2015) in the Department of Animal Science, the University of Tennessee, Institute of Agriculture, Knoxville, Tennessee. Dr. Kerro Dego’s research focuses on the prevention and control of infectious diseases of farm animals, particularly mastitis, improving dairy food safety, and mitigation of antimicrobial resistance. Dr. Kerro Dego has extensive experience in studying the pathogenesis of bacterial infections, identification of virulence factors, and vaccine development and efficacy testing against major bacterial mastitis pathogens. Dr. Kerro Dego conducted numerous controlled experimental and field vaccine efficacy studies, vaccination, and evaluation of immunological responses in several species of animals, including rodents (mice) and large animals (bovine and ovine).",institutionString:"University of Tennessee at Knoxville",institution:{name:"University of Tennessee at Knoxville",country:{name:"United States of America"}}},{id:"251314",title:"Dr.",name:"Juan Carlos",middleName:null,surname:"Gardón",slug:"juan-carlos-gardon",fullName:"Juan Carlos Gardón",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/251314/images/system/251314.jpeg",biography:"Juan Carlos Gardón Poggi received University degree from the Faculty of Agrarian Science in Argentina, in 1983. Also he received Masters Degree and PhD from Córdoba University, Spain. He is currently a Professor at the Catholic University of Valencia San Vicente Mártir, at the Department of Medicine and Animal Surgery. He teaches diverse courses in the field of Animal Reproduction and he is the Director of the Veterinary Farm. He also participates in academic postgraduate activities at the Veterinary Faculty of Murcia University, Spain. His research areas include animal physiology, physiology and biotechnology of reproduction either in males or females, the study of gametes under in vitro conditions and the use of ultrasound as a complement to physiological studies and development of applied biotechnologies. Routinely, he supervises students preparing their doctoral, master thesis or final degree projects.",institutionString:"Catholic University of Valencia San Vicente Mártir, Spain",institution:null},{id:"125292",title:"Dr.",name:"Katy",middleName:null,surname:"Satué Ambrojo",slug:"katy-satue-ambrojo",fullName:"Katy Satué Ambrojo",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/125292/images/system/125292.jpeg",biography:"Katy Satué Ambrojo received her Veterinary Medicine degree, Master degree in Equine Technology and doctorate in Veterinary Medicine from the Faculty of Veterinary, CEU-Cardenal Herrera University in Valencia, Spain. She is a Full Professor at the Department of Medicine and Animal Surgery at the same University. She developed her research activity in the field of Endocrinology, Hematology, Biochemistry and Immunology of horses. She is a scientific reviewer of several international journals : American Journal of Obstetrics and Gynecology, Comparative Clinical Pathology, Veterinary Clinical Pathology, Journal of Equine Veterinary Science, Reproduction in Domestic Animals, Research Veterinary Science, Brazilian Journal of Medical and Biological Research, Livestock Production Science and Theriogenology. Since 2014, she has been the Head of the Clinical Analysis Laboratory of the Hospital Clínico Veterinario from the Faculty of Veterinary, CEU-Cardenal Herrera University.",institutionString:"CEU-Cardenal Herrera University",institution:{name:"CEU Cardinal Herrera University",country:{name:"Spain"}}},{id:"309529",title:"Dr.",name:"Albert",middleName:null,surname:"Rizvanov",slug:"albert-rizvanov",fullName:"Albert Rizvanov",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/309529/images/9189_n.jpg",biography:'Albert A. Rizvanov is a Professor and Director of the Center for Precision and Regenerative Medicine at the Institute of Fundamental Medicine and Biology, Kazan Federal University (KFU), Russia. He is the Head of the Center of Excellence “Regenerative Medicine” and Vice-Director of Strategic Academic Unit \\"Translational 7P Medicine\\". Albert completed his Ph.D. at the University of Nevada, Reno, USA and Dr.Sci. at KFU. He is a corresponding member of the Tatarstan Academy of Sciences, Russian Federation. Albert is an author of more than 300 peer-reviewed journal articles and 22 patents. He has supervised 11 Ph.D. and 2 Dr.Sci. dissertations. Albert is the Head of the Dissertation Committee on Biochemistry, Microbiology, and Genetics at KFU.\nORCID https://orcid.org/0000-0002-9427-5739\nWebsite https://kpfu.ru/Albert.Rizvanov?p_lang=2',institutionString:"Kazan Federal University",institution:{name:"Kazan Federal University",country:{name:"Russia"}}},{id:"210551",title:"Dr.",name:"Arbab",middleName:null,surname:"Sikandar",slug:"arbab-sikandar",fullName:"Arbab Sikandar",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/210551/images/system/210551.jpg",biography:"Dr. Arbab Sikandar, PhD, M. Phil, DVM was born on April 05, 1981. He is currently working at the College of Veterinary & Animal Sciences as an Assistant Professor. He previously worked as a lecturer at the same University. \nHe is a Member/Secretory of Ethics committee (No. CVAS-9377 dated 18-04-18), Member of the QEC committee CVAS, Jhang (Regr/Gen/69/873, dated 26-10-2017), Member, Board of studies of Department of Basic Sciences (No. CVAS. 2851 Dated. 12-04-13, and No. CVAS, 9024 dated 20/11/17), Member of Academic Committee, CVAS, Jhang (No. CVAS/2004, Dated, 25-08-12), Member of the technical committee (No. CVAS/ 4085, dated 20,03, 2010 till 2016).\n\nDr. Arbab Sikandar contributed in five days hands-on-training on Histopathology at the Department of Pathology, UVAS from 12-16 June 2017. He received a Certificate of appreciation for contributions for Popularization of Science and Technology in the Society on 17-11-15. He was the resource person in the lecture series- ‘scientific writing’ at the Department of Anatomy and Histology, UVAS, Lahore on 29th October 2015. He won a full fellowship as a principal candidate for the year 2015 in the field of Agriculture, EICA, Egypt with ref. to the Notification No. 12(11) ACS/Egypt/2014 from 10 July 2015 to 25th September 2015.; he received a grant of Rs. 55000/- as research incentives from Director, Advanced Studies and Research, UVAS, Lahore upon publications of research papers in IF Journals (DR/215, dated 19-5-2014.. He obtained his PhD by winning a HEC Pakistan indigenous Scholarship, ‘Ph.D. fellowship for 5000 scholars – Phase II’ (2av1-147), 17-6/HEC/HRD/IS-II/12, November 15, 2012. \n\nDr. Sikandar is a member of numerous societies: Registered Veterinary Medical Practitioner (life member) and Registered Veterinary Medical Faculty of Pakistan Veterinary Medical Council. The Registration code of PVMC is RVMP/4298 and RVMF/ 0102.; Life member of the University of Veterinary and Animal Sciences, Lahore, Alumni Association with S# 664, dated: 6-4-12. ; Member 'Vets Care Organization Pakistan” with Reference No. VCO-605-149, dated 05-04-06. :Member 'Vet Crescent” (Society of Animal Health and Production), UVAS, Lahore.",institutionString:"University of Veterinary & Animal Science",institution:{name:"University of Veterinary and Animal Sciences",country:{name:"Pakistan"}}},{id:"311663",title:"Dr.",name:"Prasanna",middleName:null,surname:"Pal",slug:"prasanna-pal",fullName:"Prasanna Pal",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/311663/images/13261_n.jpg",biography:null,institutionString:null,institution:{name:"National Dairy Research Institute",country:{name:"India"}}},{id:"202192",title:"Dr.",name:"Catrin",middleName:null,surname:"Rutland",slug:"catrin-rutland",fullName:"Catrin Rutland",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/202192/images/system/202192.png",biography:"Catrin Rutland is an Associate Professor of Anatomy and Developmental Genetics at the University of Nottingham, UK. She obtained a BSc from the University of Derby, England, a master’s degree from Technische Universität München, Germany, and a Ph.D. from the University of Nottingham. She undertook a post-doctoral research fellowship in the School of Medicine before accepting tenure in Veterinary Medicine and Science. Dr. Rutland also obtained an MMedSci (Medical Education) and a Postgraduate Certificate in Higher Education (PGCHE). She is the author of more than sixty peer-reviewed journal articles, twelve books/book chapters, and more than 100 research abstracts in cardiovascular biology and oncology. She is a board member of the European Association of Veterinary Anatomists, Fellow of the Anatomical Society, and Senior Fellow of the Higher Education Academy. Dr. Rutland has also written popular science books for the public. https://orcid.org/0000-0002-2009-4898. www.nottingham.ac.uk/vet/people/catrin.rutland",institutionString:null,institution:{name:"University of Nottingham",country:{name:"United Kingdom"}}},{id:"283315",title:"Prof.",name:"Samir",middleName:null,surname:"El-Gendy",slug:"samir-el-gendy",fullName:"Samir El-Gendy",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRduYQAS/Profile_Picture_1606215849748",biography:"Samir El-Gendy is a Professor of anatomy and embryology at the faculty of veterinary medicine, Alexandria University, Egypt. Samir obtained his PhD in veterinary science in 2007 from the faculty of veterinary medicine, Alexandria University and has been a professor since 2017. Samir is an author on 24 articles at Scopus and 12 articles within local journals and 2 books/book chapters. His research focuses on applied anatomy, imaging techniques and computed tomography. 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