Matrix interaction \n
\r\n\t
",isbn:"978-1-83968-388-6",printIsbn:"978-1-83968-387-9",pdfIsbn:"978-1-83968-389-3",doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!1,hash:"61ec2bad4fc3f7060fd64b91fa12e82c",bookSignature:"Ph.D. Vicente Vanaclocha",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/10574.jpg",keywords:"Prevalence, Incidence, Worldwide Differences, Red Flags, Moyamoya and School Performance, Medical Treatment, Surgical Treatment, Genetic Markers, Immunologic Factors, Recommended Anesthetic Agents, Source of Intraoperative Complications, Post-Operative ICU Management",numberOfDownloads:null,numberOfWosCitations:0,numberOfCrossrefCitations:null,numberOfDimensionsCitations:null,numberOfTotalCitations:null,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"September 23rd 2020",dateEndSecondStepPublish:"October 21st 2020",dateEndThirdStepPublish:"December 20th 2020",dateEndFourthStepPublish:"March 10th 2021",dateEndFifthStepPublish:"May 9th 2021",remainingDaysToSecondStep:"3 months",secondStepPassed:!0,currentStepOfPublishingProcess:4,editedByType:null,kuFlag:!1,biosketch:"Dr. Vicente Vanaclocha is a Chief of Neurosurgery at the University Hospital of Navarra and head of Neurosurgery Service of San Jaime Hospital in Torrevieja. He has over 25 years of experience in neuro-oncology and minimally invasive surgery techniques. He is a pioneer in many areas in neurosurgery (treatment of brain tumors, Chiari Malformation, and sacroiliac joint disorders).",coeditorOneBiosketch:null,coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"199099",title:"Ph.D.",name:"Vicente",middleName:null,surname:"Vanaclocha",slug:"vicente-vanaclocha",fullName:"Vicente Vanaclocha",profilePictureURL:"https://mts.intechopen.com/storage/users/199099/images/system/199099.jpeg",biography:"Vicente Vanaclocha is Chief of Neurosurgery. Doctor of Medicine from the University of Valencia, he has over 25 years experience in neuro-oncology, minimally invasive and minimally invasive surgery techniques. Specialist in neurosurgery both nationally and internationally (including the General Medical Register of England and stay at the Groote Schuur Hospital in Cape Town, South Africa) has been Chief of Neurosurgery at the University Hospital of Navarra and head of Neurosurgery Service of San Jaime Hospital in Torrevieja. He was also associate professor of neurosurgery at the Faculty of Medicine of the University of Navarra and is a professor of neuroanatomy at the Catholic University of Valencia also serving as an editorial board member of repute.\nCurrently he is Associate Professor at the University of Valencia.",institutionString:"University of Valencia",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"7",totalChapterViews:"0",totalEditedBooks:"1",institution:{name:"University of Valencia",institutionURL:null,country:{name:"Spain"}}}],coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"16",title:"Medicine",slug:"medicine"}],chapters:null,productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},personalPublishingAssistant:{id:"297737",firstName:"Mateo",lastName:"Pulko",middleName:null,title:"Mr.",imageUrl:"https://mts.intechopen.com/storage/users/297737/images/8492_n.png",email:"mateo.p@intechopen.com",biography:"As an Author Service Manager my responsibilities include monitoring and facilitating all publishing activities for authors and editors. 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Venkateswarlu",coverURL:"https://cdn.intechopen.com/books/images_new/371.jpg",editedByType:"Edited by",editors:[{id:"58592",title:"Dr.",name:"Arun",surname:"Shanker",slug:"arun-shanker",fullName:"Arun Shanker"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"878",title:"Phytochemicals",subtitle:"A Global Perspective of Their Role in Nutrition and Health",isOpenForSubmission:!1,hash:"ec77671f63975ef2d16192897deb6835",slug:"phytochemicals-a-global-perspective-of-their-role-in-nutrition-and-health",bookSignature:"Venketeshwer Rao",coverURL:"https://cdn.intechopen.com/books/images_new/878.jpg",editedByType:"Edited by",editors:[{id:"82663",title:"Dr.",name:"Venketeshwer",surname:"Rao",slug:"venketeshwer-rao",fullName:"Venketeshwer Rao"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}]},chapter:{item:{type:"chapter",id:"19161",title:"Diagnostic of Drowning in Forensic Medicine",doi:"10.5772/19234",slug:"diagnostic-of-drowning-in-forensic-medicine",body:'The diagnostic of drowning is described in the literature as one of the most difficult in the field of forensic medicine (Piette & De letter, 2006). In fact, the external examination and the autopsy findings are in most of the cases not specific and the laboratory investigations are controversially appreciated by the scientific community. The main goal in this field is to differentiate a death by submersion from a immersion of a body. Death of a victim found in water should not always be related to drowning (Knight, 1991).
It is important to remind that the death by drowning is defined as a death due to submersion in a liquid and the mechanism in acute drowning is hypoxemia and irreversible cerebral anoxia (D.J. Di Maio & V.J.M. Di Maio 1989).
Considering the pathophysiology of human drowning, the role of mechanical airways obstruction and the washing out of alveoli surfactant as well as the shifts of fluid and electrolytes are still debated. In fact, several phases were described during the drowning process, first a breath-holding phase, followed by involuntary inspiration, gasping for air and loss of consciousness. The death is secondary of the development of cerebral hypoxia leading to irreversible brain damage. The duration of the phases is dependent on various factors, such as age, previous diseases, breath holding tolerance of the victims and the temperature of the water. Consciousness is usually lost within 3 minutes of submersion.
The inhaled water enters the alveolar spaces of the lungs and destroys the surfactant inducing pulmonary edema with the transudation of protein-rich fluid into the alveolar spaces. The surfactant washout decreases the lung compliance and ventilation-perfusion mismatch resulting to an hypoxemia secondary of non oxygenation of blood flowing through underventilated portions of the lung. A non cardiogenic pulmonary edema will result with secondary metabolic acidosis. This is the main pathophysiological mechanism of drowning and the fluid and electrolyte shifts are quite secondary.
It was stated that fresh water is hypotonic and hyponatremic relative to blood inducing, after inhalation, a movementt of water from the alveoli into the blood and movement of sodium from the blood into the alveoli. These changes induce haemodilution, hypervolemia, hypnonatremia, hyperkalemia and haemolysis (Jeanmonod et al., 1992). As the sea water is very hypertonic relative to the blood, the water movement goes from blood into the alveoli and the electrolytes (sodium, chloride, magnesium) from the alveoli into the blood. The consequences of the sea water drowning should be haemoconcentration, hypovolemia and hypernatremia. The biochemical tests that proposed to assess the diagnostic of drowning are based on these fluids and electrolytes shifts. It is during the phases where water is penetrating from the alveoli into the blood circulation that particles like diatom passing through the alveolar-capillary interface before reaching internal organs.
A vagal reflex may be also induced by inhalation of water, it will increase peripheral airway resistance with pulmonary vasoconstriction, decreased lung compliance and reduction of ventilation – perfusion ratios (Ornato, 1986).
An intense stimulation of nerve endings at the skin, the mucosa of the ear drum, the pharynx or the larynx by cold water can lead to a cardiac reflex arrest. It was assumed that 10% of the drowned humans die after laryngospasm or breath-holding without actually aspirating fluid (Ludes & Fornes, 2003). A discussion was also hold about the volumes of inhalated water and the effect on the circulation. In drowning, the inhalated volume of water can range, from relatively small to very large. It has been showed that small amount of water, particularly cold water, may induce vaso vagal reflex or cardiac arrest reflex. When great amounts of water are inhalated and pass through the alveolar-capillary interface and enter the circulation, the phenomenon of destruction of surfactant and of the alveoli architecture leads to asphyxia. During the entering of water into the blood stream, the diatoms present in the drowning fluid may reach the internal organs.
To establish the diagnosis of drowning it is of particular importance to correlate informations about the circumstances preceding the death, the past medical history of the victim if known, the circumstances of the body recovery from the water, the external examination, the autopsy findings and the results of the complementary analyses (histologic, biochemical, toxicological analyses and diatom test).
The majority of the autopsy findings are related to asphyxia and have no specific link to drowning. The signs of drowning depend on the delay in recovering the body and on the development of the putrefaction phenomenon which alter the positive signs of drowning. One of the signs of drowning would be large amounts of froth present around nostrils and mouth in freshly drowned bodies. This froth is also present in the upper and lower airways. Froth can also be observed in cases of edema of left ventricular failure but in drowning cases the volume of froth is generally much more abundant than in other origins.
It is admitted that lung weights are higher in drowning cases but it was shown that normal weights are possible in the drowning cases after cardiac arrest reflex or vaso vagal reflex. After water inhalation, the lungs may be over inflated, filling the thoracic cavity, generally water logged referred to as “emphysema aquosum”. So the surfaces of lungs have a marbled appearance with dark red areas linked with collapsed alveoli, interspersed with more aerated tissues areas. The fluid is trapped in the lower airways and blocks the passive collapse of the bronchi that normally occurs after death. Subpleural bullae of emphysema, sometimes hemorrhagic may be found and are related to the rupture of the alveolar walls (Pounder, 2005). Even if these signs are mostly evocating of drowning, none of them is pathognomonic of water inhalation.
The body having sunk to the bottom of the site of drowning, will show a pattern of post mortem injuries such as post-mortem abrasion over the forehead, the prominent points of the face, the anterior trunk, the backs of the hands and the fronts of the lower legs. Injuries may also inflicted by passing watercraft in navigable waters by stumbling against rocks or by animal activities. Accidental or suicidal injuries due to the way the person falls or enters into the water may also be observed. Post-mortem injuries linked to the way of recovery of the body using ropes and hooks can also be seen. These kinds of post-mortem injuries can mimic ante-mortem wounds and the differentiation between ante and post-mortem injuries is quite difficult because of the lack of the usual criteria of ante-mortem wounds.
It can also be found sand, silt, seashells and weeds in the airways, lungs, stomach and duodenum of drowned victims. If this material is fund in abundance within the alveoli, it can be related to an immersion during life as long as it concerns a freshly drowned body. This material may also enter the upper airways during the post-mortem immersion period and it is possible that small quantities may enter the oesophagus and stomach but it is unlikely that it will reach the alveoli to any significant extent if the post-mortem submersion is short.
One of the sign of immersion is skin maceration becoming visible after various time interval depending on the temperature of the immersion water. The skin becomes wrinkled, pale and sodden like a “washer woman’s skin”. These changes appear at the finger tips, palms, backs of the hands, and later, the soles. The next step is the detachment of the thick keratin of hands and feet which pull off in “glove and stoking fashion”. Nails and hair become loosened after a few days. Other signs of immersion are cutis anserine and post-mortem distribution of hypostasis. The presence of mud, silt or sand on the body was described but has no diagnostic value.
The microscopic investigations must be performed on all the organs of non putrefied bodies in the aim to make the difference between a death by drowning and other causes of death. The lung examinations can show over-distension of the alveoli, thinning of the alveolar septa and compression with narrowing of the capillary network (Pounder, 2005).
The modifications in lungs are heterogeneous distributed and multiple sections must be performed to assess the diagnostic. In fact the microscopic appearance may be entirely normal in some part of the lungs.
Several staining techniques must be performed such as the staining for elastic fibers (orcein ) and reticulin fibers (Fornes et al., 1988 ; Ludes & Fornes, 2003). The examination of other organs (brain, heart, liver) shows none specific histological changes indicative of hypoxia such as acute congestion and swelling of the capillary endothelia.
The chemical changes in plasma after drowning were based on the fluid and electrolyte shifts after the penetration of either sea or fresh water in the alveoli and in the blood stream (Modell & Davis, 1969). It was proposed the measurement of the specific gravity of blood, of the concentration of sodium, chloride and potassium. For the electrolytes, the diagnosis of drowning was based on changes of these electrolytes between the blood samples taken from the right versus left ventricle (Bray, 1985; Couteselinis & Boukis 1976; Karkola & Neittaanmaki, 1981). Such electrolyte shifts were described in many other causes of death and do not provide reliable evidence of drowning.
A special mention must be made for the blood strontium analysis. The toxicological analysis are performed to show the presence of medicaments or alcohol, taken before death in suicidal or accidental conditions and to determine the serum level of strontium which is described as a good parameter of drowning in sea water (Piette & Timperman, 1989). In case of fresh water drowing, the water concentration of strontium must be higher than the serum concentration to be a valuable parameter in favour of drowning.
Authors such as Kane et al. (1996, 2000) and Nübel et al. (1997) proposed the detection by molecular biology techniques of the 16S rRNA subunits of ribosomal RNA for plankton detection in tissues samples indicating an active water inhalation and may assess the diagnostic of drowning. According to these authors, the sequence comparison of the variable regions of 16S rRNA could provide sufficient information to allow the discrimination of both close and distant phylogenetic relationships.
Abe et al. (2003) and Suto et al. (2003) proposed the detection of chlorophyll-related genes of Euglena gracilis and Skeletonema costatum to identify plankton in the victim’s tissues. It is important to emphasize that these methods give only qualitative results (He et al., 2008) but the quantitative approach can only be achieved by the diatom test which may also give an indication of the site of drowning.
In fact, diatoms can be considered as particles present in the submersion water which are inhalated during drowning and once in the blood stream which reach the closed organs. Under strict extraction and identification conditions, these particles are good markers of drowning.
Diatoms are unicellular algae belonging to the class of bacillariophycae which includes more than 15 000 species living in fresh, brakish or sea water. The skeleton of these algae is called a frustule which is constituted by two valves fitting together to enclose the cytoplasm (Ludes & Fornes, 2003) and made of hard silice.
Due to this hard silicaceous skeleton, diatoms can be recovered from putrefied or burnt tissues by either enzymatic or acid digestion (Ludes et al., 1994). The identification of these algae is based on the structure of their valves showing different symmetry allowing the distinction of two main groups namely the centric diatoms and the elongated or pennate diatoms.
After a long period of time where the use of the diatom test was very controversial due to false positive results linked to the presence of diatoms in closed organs of non drowned victims (Foged, 1983; Gylseth & Mowe, 1979; Schellmann & Sperl, 1979; Schneider, 1980, 1990; Schneider & Kolb, 1969), it was stated by several authors that under strict defined analytical conditions this test could discriminate between drowning and none drowning cases (Auer & Möttönen, 1988 ; Neidhard & Greedyke, 1967; Peabody, 1977; Pollanen, 1997,1998; Pollanen et al., 1997). Auer & Möttönen (1988) were one of the first authors to propose that 20 diatoms per microscope slide obtained from lung samples should be a sufficient concentration to exclude false positive due to contaminations. We also proposed qualitative and quantitative criteria for a positive drowning diagnostic with the diatom test.
For us, an analysis will be considered as positive when at least 20 diatoms are identified per 100 µl of a pellet sediment extracted from a 2 g lung sample and the identification of more than 5 complete diatoms (with exclusion of fragments) per 100 µl of a pellet sediment extracted from a 2 g tissue samples such as brain, kidney, liver and bone marrow. Bone marrow is described as a sanctuary organ and if diatoms reach this tissue, the diagnostic of drowning could be assessed.
In controlled samples belonging to non drowned victims, we newer find a number of diatoms above the fixed criteria. When diatoms were found in closed organs of drowned victims, the results in lung samples were in each case also above the 20 algae per 100 µl pellet. To assess the diagnostic of drowning, it is of high importance to perform a qualitative analysis of the found diatoms and the comparison of the diatoms present in the closed organs and the microflora of the presumed site of drowning.
In this aim, water samples must be collected at the drowning site (two samples of 100 ml) as well as algae scraped from stones present in the water.
The samples are disposed in clean containers and the extraction and identification protocols on water and tissue samples were described by our group (Ludes et al., 1999). All reagents and glass containers must be checked for the absence of diatoms before use, and contamination from exogenous diatoms must be avoided by using diatom-free water and by protecting the organs during autopsy from the clothes of the victims and from the skin surface.
At each step of the analyses and the identification of diatoms, a potential contamination must be considered. This test cannot be proposed to assess the diagnostic of drowning in bathtub or in water containing very few algae, for example in iced water.
If water samples are not available, it is possible to compare the diatoms found in the organs with data collected in the rivers by a continuous water monitoring which can be set up by, for example, the Agencies in charge of the survey of water quality.
We set up a continuous water monitoring of the main rivers of our area (Ludes et al., 1996). Seasonal variations of the concentration of diatoms and the diatom profile are determined at a given month by the five most frequent species. The relative abundance of each diatom may also vary along the course of the river. So, the site of drowning may be determined by comparison between the water microflora with the diatoms found in the lungs. In fact, diatoms of more than 50 – 60 µm in size rarely pass the alveoli-capillary barrier even after the rupture of the alveoli by the inhalation of water.
The diagnostic of drowning can be achieved when the qualitative analysis shows that the algae found in the organ belongs to the water microflora and the quantitative criteria are fulfilled (Hendey, 1973, 1980 ; Ludes et al., 1999).
The diagnostic of drowning may be achieved after having considered all the forensic investigations performed in those cases, i.e: external examination, autopsy findings, histological and toxicological analysis, blood strontium determination, biochemical analysis and diatom test. The diatom test was still considered controversial by the by the literature but we defined qualitative and quantitative criteria which could exclude false positive results. It is of particular interest in case of putrefied bodies where the other investigations have failed.
The scenario of the modern era of interdisciplinary research between the natural sciences, including mathematics and theoretical physics, biophysics, chemistry, colloid science, and biochemistry, among others, along with advances in computational sciences, makes possible the manipulation of matter on a molecular scale through nanotechnology and bionanotechnology. This interdisciplinary research is able to study nanosystems made of phospholipids can be self-assembled spontaneously. One of the most studied cases in recent years is the self-assembly of liposomes.
\nThe liposomes, vesicular systems based on lipids, in the last decades have increased their potential of their applications in various scientific disciplines and extensively been used as carriers for numerous molecules in cosmetic and pharmaceutical industries and medical and agricultural fields due to their high capacity and efficiency in drug delivery [1, 2]. For purposes in drug delivery systems, the liposomes are suitable candidates, due to their multiple properties that make them unique in these processes, such as biocompatibility, biodegradability, low toxicity, ability to modify the pharmacokinetic profile of the drug loaded, etc. [3]. In this way, liposomes have been successfully applied in the delivery of nucleic acid systems [4], in the intestinal lymphatic delivery [5], for enhanced oral delivery [6]. Allen and Cullis documented that these advances have led to numerous clinical trials in such diverse areas as the delivery of anticancer, antifungal, and antibiotic drugs; the delivery of gene medicines; and the delivery of anesthetics and anti-inflammatory drugs [7]. Despite the innumerable applications and advances of the drug delivery of liposomes, it presents enormous challenges still. For instance, He et al. argued that by modulating the compositions of the lipid bilayers and adding polymers or ligands, both the stability and permeability of liposomes can be greatly improved for oral drug delivery [8]. In order to achieve direct, effective, and selective delivery of the liposome itself, a deeper understanding of the mechanisms for interaction between the water/ligands/polymers and the liposome is required [9]. It has been reported in different sources that the mechanism of liposome formation is based on the unfavorable interactions occurring between amphiphilic compounds and water molecules, where the polar head groups of phospholipids are subjected to the aqueous phases of the inner, the hydrophobic hydrocarbon tails are associated with the bilayer, and spherical core shell structures are formed [10, 11, 12].
\nIn addition to a large number of experiments that have been dedicated to the study of the mechanism of liposome formation, computational studies have made important contributions to the subject. These methodologies have been shown to be very effective in knowing in a very specific way the role played by molecular interactions in complex systems such as liposome self-assembly. Nowadays, it is increasingly common for research protocols to include a computational analysis prior to carrying out the experimental stage. Different techniques of computer modeling can facilitate quantitative understanding of experimental observations and secure fundamental interpretation of underlying phenomena [13]. For instance, Thota and Jiang in an interesting review documented that with the use of simulation methods, it is possible to elucidate the mechanisms of drug loading/release, which are indispensable in the rational screening and designing new amphiphiles for high-efficacy drug delivery [14].
\nMost of the computational studies for drug delivery use molecular dynamics (MD) or Monte Carlo (MC) simulations to obtain structural and dynamic information. In the case of MD, for example, the study of interactions is carried out considering all atoms explicitly commonly. These interactions are evaluated by analytical functions that form the so-called force fields. The level of precision of a simulation with MD relies on the force field used. There are traditional force fields such as CHARMM [15], AMBER [16], and OPLS [17] used to study the self-assembly of complex systems such as liposomes. Due to the scheme of evaluation of the interactions, which is through both the intra- and intermolecular forces, the MD simulations are slow and computationally expensive when the number of atoms is very large. This limitation makes it almost impossible to reach the time scales (of the order of the microseconds and beyond) where the thermodynamic observables have greater physical significance. For this reason, this technique turns out to be little viable to study the systems planned in this work, although it is important to point out that diverse groups use it with very successful results.
\nModeling technique alternatives to MD have been developed with the aim of overcoming the bottleneck when it is required to study systems composed of a large number of atoms, as well as the issue of length time scales. One of those techniques is the dissipative particle dynamics (DPD). The main philosophy of the DPD model is based on the so-called coarse-grained (CG) models. These models allow to diminish considerably the number of atoms of a given system, grouping a set of atoms, which can be a functional chemical group, into a single interaction site called a bead, always trying to maintain the physics of the correct model. This size reduction facilitates larger simulations of larger ones with larger systems, allowing a more efficient exploration of phase space, but losing an atomistic description. Despite the success of the DPD technique, very few groups have used it to obtain detailed information on various aspects of self-assembly of liposomes. Goicochea et al. used DPD for understanding the behavior of biopolymer brushes coating drug-carrying liposomes in an aqueous environment [18]. Shen et al. with the help of DPD simulations, they explored how the tethered PEG polymers will affect the membrane wrapping process of PEGylated liposomes during endocytosis [19]. Yamamoto et al. used DPD simulations to study the spontaneous vesicle formation of amphiphilic molecules in aqueous solution [20].
\nAnother great advantage of the modeling methods such as DPD is to be able to incorporate their algorithms into powerful supercomputing technologies such as graphic processing units (GPUs). The combination of hardware and software with DPD has allowed us to explore systems at time scales, which were unattainable a few years ago. For instance, Wu et al. [21] presented a GPU-accelerated DPD with parallel cell-list updating. On the other hand, Nguyen et al. [22] achieve a speedup of up to 9.5x in the DPD simulations in the well-known LAMMPS code using GPUs. Phillips et al. [23] also achieve important speedup of DPD using GPUs but the HOOMD-blue code. Other important implementations of DPD in GPUs can be found in Refs [24, 25]. Our group has been a pioneer in the use and application of DPD methodologies using GPUs. We have developed a simulation code called SIMES [26] (which is an acronym for simulation at mesoscopic time scale), a production DPD software designed and implemented specifically to run on GPUs. SIMES can simulate a broad spectrum of complex systems, including the self-assemblies of liposomes. Readers interested in going deeper into this topic can consult our contributions in the following references [27-30].
\nRecently, in a previous work of our group, we have studied the stability of structures and the efficiency of the encapsulation of capsaicin, as well as the internal and superficial distribution of capsaicin and chitosan inside of a nanoliposome, constituted by lecithin and coated with a shell of chitosan by DPD simulations [31]. We found that thermodynamic properties, such as the potential mean force, show that the interaction between capsaicin and chitosan polymers is very weak compared to that with lecithin. Besides, an association between capsaicin and chitosan in presence of lecithin is not likely to occur. In this work, we managed to combine the power of the DPD method with the power of the GPUs. The simulations reported have been the longest so far by regarding the encapsulation of capsaicin by liposomal nanoparticles.
\nContinuing with our research computational about self-assembly of molecular components for the design of bionanomaterials focused on drug delivery applications, the objective of this chapter is to show the advantages offered by the DPD technique to study the self-assembly mechanism of liposomes. As a specific case study, we show the formation of liposomes loaded with capsaicin. Capsaicin, a lipophilic drug, is the pungent vanilloid compound in spicy chili peppers. We chose capsaicin because it is approved as a drug for the treatment of chronic pains [32]. It is also an excellent candidate to be transported by natural polymers such as chitosan [33]. In addition, it is widely recognized in the treatment of urological disorders and the control of satiety and obesity [34], among other important applications.
\nThe simulation of DPD is one of the most viable methods to study thermodynamic and structural properties of biophysical processes at mesoscopic time and length scales, unlike the simulation with MD, where insights at microscopic time and length scales can achieved. This great advantage of DPD simulation is that the DPD models are built under the coarse-grained scheme, which is obtained from the description of all-atom of a given system. This coarse-graining parameter plays a vital role and has significant impact on the speed of simulation [35]. This feature makes the DPD simulations cheap and very fast computationally speaking.
\nThe theoretical foundations of the DPD method are documented elsewhere. Readers interested can consult the following references [36-40] for more specific details. In this chapter, we will mention in a very summarized way the main mathematical aspects that govern the equations of this methodology. The DPD was introduced by Hoogerbrugge and Koelman in 1992 [41] for simulations of hydrodynamic phenomena. It was subsequently modified by Español and Warren in 1995 [42] who introduced the fluctuation-dissipation theorem in the original algorithm to add the frictional and stochastic forces. Therefore, a coupling between the statistical mechanic laws of the beads and Gibbs canonical ensemble is given, favoring the calculation of thermodynamic properties at longer time and length scales. Similar to molecular dynamics, the time evolution of each DPD particle can be calculated by Newton’s second law:
\nwhere \n
where \n
where \n
Technically, DPD simulation differs from MD simulation in two respects. First, the conservative pairwise forces between DPD particles are soft-repulsive, which makes it possible to extend the simulations to longer time scales. Second, the scheme to maintain constant temperature in DPD is implemented in terms of dissipative as well as random pairwise forces such that the momentum is locally conserved, which results in the emergence of hydrodynamic flow effects on the macroscopic scale.
\nFrom the molecular point of view, a liposome, which is usually amphiphilic molecules, contains polar heads and hydrophobic hydrocarbon tails. This molecular structure is ideal to build a DPD model, since the polar part can be grouped in a single bead, while the hydrophobic part can be grouped in another bead. The liposome presented in this work is constituted by lecithin, which has two long hydrocarbon chains, is a major component of lipid bilayers of cell membranes and is a natural and biological amphiphile. Lecithin is a natural lipid mixture of phospholipids used frequently for the preparation of various nanosystem delivery vehicles, such as liposomes [9, 10]. According to the molecular structure of lecithin, this can be separated into three different beads that correspond to the head, neck, and tail groups, respectively. A schematic illustration of the fundamental self-assembly of the structure of a liposome adopted in this chapter is shown in Figure 1. In Figure 1A, a detailed molecular description at all-atom level for the lipid is shown; a mapping in the bead representation of the atomistic structure is also shown. Figure 1B is depicting the coarse-grained model of lecithin used in this work with its three fundamental parts. A snapshot of configurational structure of the liposome constructed based on lecithin is shown in Figure 1C. From this representation, the yellow spheres represent the hydrophobic part of the lecithin, while the red spheres represent the hydrophilic part.
\nOn the other hand, we show the coarse-grained model of capsaicin, our target as case of study in drug delivery applications. The molecular structure of capsaicin is also ideal for building a DPD model. Similar to lecithin, the capsaicin can be grouped into three groups or beads, i.e., one bead for the head, neck, and tail groups, respectively. Figure 1A shows the coarse-grained model for capsaicin.
\nFinally, in Figure 1A, we show the coarse-grained model of water. This model in DPD simulation is already well known; for example, a bead DPD can group up to three water molecules. This model allows to conserve the thermodynamic properties of water.
\nSchematic illustration of computational models used for self-assembly of liposomes, from atomistic to mesoscopic representation. (A) Example of a lipid in a representation of all-atom. The computational model of coarse grain is constructed by grouping certain functional groups that form the fundamental molecular structure. In this case, the 35 atoms of the molecular structure of the phospholipid are grouped into only five sites or pseudo-atoms of coarse-grained type. (B) Example of our coarse-grained model of a lipid. The red spheres represent the hydrophobic group, while yellow spheres represent the hydrophilic group. (C) A representation of a liposome showing the inner of aqueous nucleus.
Mesoscopic simulations were performed to study the self-assembling of liposomes. The capsaicin was used as an example of drug delivery applications. For this end, the simulations presented in this work were made under canonical ensemble, where N (the total particle number), V (volume), and T (temperature) are kept constant. For this type of simulations at constant temperature, some parameters must be specified. In this work, in all the simulations, we use dimensionless units or DPD units, i.e., the temperature and density were of T = 1.0, ρ = 3.0, respectively, the mass of all particles is \n
\n | L1 | \nL2 | \nL3 | \nW | \n
---|---|---|---|---|
L1 | \n78.0 | \n80.0 | \n95.0 | \n89.0 | \n
L2 | \n80.0 | \n78.0 | \n86.0 | \n93.0 | \n
L3 | \n95.0 | \n86.0 | \n78.0 | \n101.0 | \n
W | \n89.0 | \n93.0 | \n101.0 | \n78.0 | \n
Matrix interaction \n
The nomenclature used in this table goes as follows: the symbols L1, L2, and L3 represent the head, neck, and tail beads of lecithin, while that W symbol represents the bead corresponding the beads of water.
\nOn the other hand, the parameters of intramolecular interactions in lecithin are \n
For the study of self-assemblies of liposomes for drug delivery applications, three different processes are presented. First, we analyze lipid self-assembly. Second, self-assembly of capsaicin is presented. Third, we end with the analysis of self-assembly of capsaicin by lecithin. The formation of nanoliposomes was explored by varying the concentration of both lecithin and capsaicin molecules separately. These simulations were analyzed through its density maps. This property is very useful for refining structures or adjusting molecular models; besides, the density maps allow us to spatially inspect the average location of atoms of interest during a simulation [46].
\nTo explore the self-assemblies of lipids, the density maps of five different processes were obtained. In these processes, the effect of lipid concentration was explored. Figure 2 shows the snapshots of each of these processes. We observe that in Figure 2A, with a concentration of 1000 lipid molecules, the self-association of lipids is practically negligible. Only a slight formation of small clusters is perceived, similar to the density maps of very diluted solutions. By increasing the concentration of lipids to double, that is, to 2000 molecules as can be seen in Figure 2B, we observe that the association increases with respect to the previous case of 1000 lipid molecules. However, it is not yet a sufficient concentration to detect any indication of the process of self-assembly of lipids. Figure 2C shows the case for a concentration of 3000 lipid molecules. In this system, the formation of very faint regions is observed. The presence of these very light red dots is an indication that the lipids begin to self-assemble. To verify that the lipid molecules are indeed self-assembling, a simulation with 4000 lipid molecules was carried out. The result is shown in Figure 2D. As we expected, the formation of the red dots increases, and they begin more notoriously. To observe the stability in the formation of lipids, a simulation with 5000 molecules was carried out. At this concentration, the lipid association is already clearly observed. A picture of this system is seen in Figure 2E. The red regions show that the self-assembly process has already taken place. These are the optimal conditions in terms of concentration for the process of self-assembly of lipids to be carried out. For higher concentrations of lipids, greater than 5000 molecules, we will find a saturation of the system, which is an amount sufficient for the formation of a nanoliposome. An animation video of each of these systems can be found in the supplementary material section of this work.
\nDensity maps for exploring the self-assembly of lipids. (A) 1000 lipid molecules, (B) 2000 lipid molecules, (C) 3000 lipid molecules, (D) 4000 lipid molecules, and (E) 5000 lipid molecules. Below each density map, a snapshot of the final configuration is displayed using the same color code as Figure 1. The solvent molecules are not shown for clarity purposes.
We conducted an exploration to study the self-assembly of capsaicin. For this purpose, we carried out four separate numerical experiments. The results of these simulations are found in Figure 3, analyzed through their density maps. These maps show regions of high and low density basically. The regions of low density are the ones that predominate the colors black and purple, while the regions of high density are those that are identified in lighter shades such as red or yellow. Figure 3A shows the density map for a system composed of 250 capsaicin molecules. Due to the low concentration of capsaicin molecules, the black and purple regions predominate, which are located in a density range between 0.02 and 0.04. In the second numerical experiment, we doubled the concentration of capsaicin molecules. Figure 3B shows the density maps for this system composed of 500 capsaicin molecules. The presence of regions in all reddish is notorious, which oscillates between 0.08 and 0.12 in density. However, purple regions are still observed, which indicate that this concentration of capsaicin molecules is not yet enough to have a homogenous system. Figure 3C shows the density map for a system composed of 750 capsaicin molecules. Unlike the density map of Figure 3B, the presence of purple areas is already minimal, while the presence of high-density areas is more consolidated. A higher density of capsaicin molecules is necessary to completely eliminate the purple areas. This can be seen in Figure 3D, which shows the density map for a concentration of 1000 capsaicin molecules. Note in this case how the purple areas have completely disappeared. The entire density map is in a homogenous system with a density that ranges between 0.2 and 0.25. This concentration of capsaicin molecules is optimal for use in the process of formation of nanoliposomes.
\nDensity maps for exploring the self-assembly of capsaicin. These snapshots correspond to equilibrated systems composed of (A) 250 capsaicin molecules, (B) 500 capsaicin molecules, (C) 750 capsaicin molecules, and (D) 1000 capsaicin molecules.
We have documented that nanoliposomes are effective for the drug delivery process. A strategic requirement for the supply of medicines to a specific site has driven the development of active drug targeting techniques [47]. In this way, to take advantage of the effective potential of the liposome technology and the impact on its formulations, it is necessary to know the optimal conditions in terms of concentration of each of the components that are required for an optimal application of any given drug. To explore the self-assembly of capsaicin by means of lecithin liposomes, we conducted four numerical experiments. The number of capsaicin molecules used in these four experiments was 250, 500, 750, and 1000 corresponding to a concentration of 30, 61, 92, and 123 mM, respectively, and the concentration of lecithin remained constant with 5000 molecules in the four cases. Figure 4 shows the density maps obtained for these four cases. Figure 4A–D shows the density maps for a system loaded with 250, 500, 750, and 1000 capsaicin molecules. For all four we observed a homogeneous formation of the liposome. The high-density regions are representing the region formed by the lipid membrane, which in this case is presented as the region with the highest concentration (in yellow) in the four cases. In addition, in the four cases, they present a redder region in the center of the map; this indicates that the density of the lipids is lower, as expected. Density maps also show that the nanoliposome is stable for 4.8 μs of simulation, since the lecithin molecules do not spread throughout the simulation box nor do they collapse in the aqueous core to form a micelle; this was possible thanks to the previous analysis where we found the optimal concentration of lecithin molecules in the self-assembly process.
\nIn our previous study [31], we observed the formation of a nanoliposome based on lecithin was carried out with approximately 7000 molecules of lecithin, but in that study, the presence of natural polymers such as chitosan was included, a component that is not present in these simulations.
\nDensity maps for exploring self-assembly of capsaicin by lecithin nanoliposomes. These snapshots correspond to equilibrated systems composed of (A) 250 capsaicin molecules, (B) 500 capsaicin molecules, (C) 750 capsaicin molecules, and (D) 1000 capsaicin molecules.
Derived from the results obtained in the density maps of Figure 4, we proceeded to make an analysis on the encapsulation efficiency (EE). It has been reported that both size and EE are key variables in the preparation of liposomes [47]. However, these variables are difficult to control at the laboratory level even with sophisticated experimental equipment. Computational tools such as DPD are ideal for guiding experiments. The size and EE for the system analyzed in this work are show in Table 2. EE is obtained from the following equation: EE = ((CapsT − CapsF)/CapsT) × 100, where CapsT is the total concentration of capsaicin in the system and CapsF is the free capsaicin in solution. The percentage of EE for the capsaicin obtained in this work was of 96%, which is very close to that reported experimentally of 92% [48], besides, is very similar to the obtained percentage in our previous study, where the presence of chitosan polymer was included [31].
\nConcentration of capsaicin (Mn) | \nSize (nm) | \n± (nm) | \nEE (%) | \n± (%) | \n
---|---|---|---|---|
30 | \n17.95 | \n0.46 | \n96.27 | \n0.69 | \n
61 | \n18.01 | \n0.61 | \n96.93 | \n0.36 | \n
92 | \n17.90 | \n0.21 | \n96.81 | \n0.35 | \n
123 | \n17.97 | \n0.16 | \n96.94 | \n0.25 | \n
Mean size of the nanoliposome and encapsulation efficiency (EE) as a function of concentration of capsaicin.
In this chapter, simulations at the mesoscopic scale were used to systematically study the self-assembly of liposomes and their potential applications toward the delivery of drugs. The advantages offered by the mesoscopic methods such as DPD over other simulation methods such as MD were exposed; additionally, the acceleration that the DPD simulations have achieved through the new GPUs technologies was also covered. With these technologies, we show simulations in the microsecond scale with a system composed of 150,000 particles in the process of self-assembly of liposomes. We show the optimal conditions for the formation of liposomes in function of the lecithin concentration to achieve a good capsaicin encapsulation. Our percentage of EE obtained is quite acceptable with respect to experimental measurements and simulation studies of similar systems.
\nThis work was supported by CONACYT [Project FON.INST./44/2016] and SIEA-UAEM [Project 4728/2019CIB]. All simulations reported in this work were performed at the Supercomputer OLINKA, located at the Laboratory of Molecular Bioengineering at Multiscale (LMBM), at the Universidad Autónoma del Estado de México.
\nThe authors declare that they have no competing interests.
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After obtaining a Master's degree in Mechanical Engineering, he continued his PhD studies in Robotics at the Vienna University of Technology. Here he worked as a robotic researcher with the university's Intelligent Manufacturing Systems Group as well as a guest researcher at various European universities, including the Swiss Federal Institute of Technology Lausanne (EPFL). During this time he published more than 20 scientific papers, gave presentations, served as a reviewer for major robotic journals and conferences and most importantly he co-founded and built the International Journal of Advanced Robotic Systems- world's first Open Access journal in the field of robotics. Starting this journal was a pivotal point in his career, since it was a pathway to founding IntechOpen - Open Access publisher focused on addressing academic researchers needs. Alex is a personification of IntechOpen key values being trusted, open and entrepreneurial. Today his focus is on defining the growth and development strategy for the company.",institutionString:null,institution:{name:"TU Wien",country:{name:"Austria"}}},{id:"19816",title:"Prof.",name:"Alexander",middleName:null,surname:"Kokorin",slug:"alexander-kokorin",fullName:"Alexander Kokorin",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/19816/images/1607_n.jpg",biography:"Alexander I. Kokorin: born: 1947, Moscow; DSc., PhD; Principal Research Fellow (Research Professor) of Department of Kinetics and Catalysis, N. Semenov Institute of Chemical Physics, Russian Academy of Sciences, Moscow.\r\nArea of research interests: physical chemistry of complex-organized molecular and nanosized systems, including polymer-metal complexes; the surface of doped oxide semiconductors. 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I received a B.Eng. degree in Computer Engineering with First Class Honors in 2008 from Prince of Songkla University, Songkhla, Thailand, where I received a Ph.D. degree in Electrical Engineering. My research interests are primarily in the area of biomedical signal processing and classification notably EMG (electromyography signal), EOG (electrooculography signal), and EEG (electroencephalography signal), image analysis notably breast cancer analysis and optical coherence tomography, and rehabilitation engineering. I became a student member of IEEE in 2008. During October 2011-March 2012, I had worked at School of Computer Science and Electronic Engineering, University of Essex, Colchester, Essex, United Kingdom. In addition, during a B.Eng. 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