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",isbn:"978-1-83969-648-0",printIsbn:"978-1-83969-647-3",pdfIsbn:"978-1-83969-649-7",doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!0,hash:"b05fa99bbd2c8e02d48dc740c0efbf9c",bookSignature:"Dr. Amjad Zaki Almusaed",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/10986.jpg",keywords:"Environmental Housing, Ecological Housing, Climate Change and Housing, Low Energy Housing Design, Recycling Materials, Human Sheltering, Human Mobility, Accessibility and Housing Units, Economic Theory and Housing, Land Administration in Habitat Zone, Decentralization in Housing Area, Private Housing",numberOfDownloads:null,numberOfWosCitations:0,numberOfCrossrefCitations:null,numberOfDimensionsCitations:null,numberOfTotalCitations:null,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"March 5th 2021",dateEndSecondStepPublish:"April 2nd 2021",dateEndThirdStepPublish:"June 1st 2021",dateEndFourthStepPublish:"August 20th 2021",dateEndFifthStepPublish:"October 19th 2021",remainingDaysToSecondStep:"17 days",secondStepPassed:!0,currentStepOfPublishingProcess:3,editedByType:null,kuFlag:!1,biosketch:"Dr. Almusaed is focused on sustainability in architecture and urban planning and design. He has carried out a great deal of research and technical survey work as well as several studies in the aforementioned areas with over 170 published international academic works in different languages.",coeditorOneBiosketch:null,coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"110471",title:"Dr.",name:"Amjad",middleName:"Zaki",surname:"Almusaed",slug:"amjad-almusaed",fullName:"Amjad Almusaed",profilePictureURL:"https://mts.intechopen.com/storage/users/110471/images/system/110471.png",biography:"Amjad Almusaed was born in 1967. He holds a PhD degree in Architecture (Environmental Design) from Ion Mincu University, Bucharest, Romania. He completed postdoctoral research in 2004 on sustainable and bioclimatic houses, from the School of Architecture in Aarhus, Denmark. His research expertise is sustainability in architecture and urban planning and design. He has carried out a great deal of research and technical survey work, and has performed several studies in the above-mentioned areas. He has edited many international books and is an active member of many worldwide architectural associations. He has published more than 170 international academic works (papers, research, books, and book chapters) in different languages.",institutionString:"Jönköping University",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"8",totalChapterViews:"0",totalEditedBooks:"9",institution:{name:"Jönköping University",institutionURL:null,country:{name:"Sweden"}}}],coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"11",title:"Engineering",slug:"engineering"}],chapters:null,productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},personalPublishingAssistant:{id:"259492",firstName:"Sara",lastName:"Gojević-Zrnić",middleName:null,title:"Mrs.",imageUrl:"https://mts.intechopen.com/storage/users/259492/images/7469_n.png",email:"sara.p@intechopen.com",biography:"As an Author Service Manager my responsibilities include monitoring and facilitating all publishing activities for authors and editors. From chapter submission and review, to approval and revision, copyediting and design, until final publication, I work closely with authors and editors to ensure a simple and easy publishing process. I maintain constant and effective communication with authors, editors and reviewers, which allows for a level of personal support that enables contributors to fully commit and concentrate on the chapters they are writing, editing, or reviewing. I assist authors in the preparation of their full chapter submissions and track important deadlines and ensure they are met. I help to coordinate internal processes such as linguistic review, and monitor the technical aspects of the process. As an ASM I am also involved in the acquisition of editors. 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Theophanides",coverURL:"https://cdn.intechopen.com/books/images_new/1591.jpg",editedByType:"Edited by",editors:[{id:"37194",title:"Dr.",name:"Theophanides",surname:"Theophile",slug:"theophanides-theophile",fullName:"Theophanides Theophile"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}]},chapter:{item:{type:"chapter",id:"49939",title:"Utilization of Ground-Penetrating Radar and Frequency Domain Electromagnetic for Investigation of Sewage Leaks",doi:"10.5772/62156",slug:"utilization-of-ground-penetrating-radar-and-frequency-domain-electromagnetic-for-investigation-of-se",body:'Water pollution is the contamination of bodies of water such as aquifers, lakes, ponds, rivers and oceans. This contamination occurs due to direct or indirect discharge of pollutants into the water bodies, without a suitable treatment to remove harmful compounds (pollutants may simply be defined as substances added to the environment that do not belong there). A substantial proportion of water and environmental contaminants are due to leaks from underground sewage pipeline systems in rural, urban and industrial areas, since any sewage pipeline system deteriorates over time, developing cracks and joint defects. Therefore, if sewage pipeline systems are not maintained properly, it is only a matter of time before the sewage leaks out and contaminates the surrounding groundwater and surface water.
Here, we suggest detecting sewage leaks from pipeline systems using two orthogonal active remote-sensing methods: (I) ground-penetrating radar (GPR) and (II) frequency domain electromagnetic (FDEM). Our hypothesis is that GPR and FDEM screening, which creates subsurface images around and along pipeline systems, will enable the extraction of residual signals and the detection of meaningful leaks. Like most complex near-surface detection missions, detection of sewage leaks in an urban environment requires a professional understanding of the regional setting, from geomorphological, environmental and engineering perspectives.
Advances in remote-sensing technologies now enable their use to identify leakage that is potentially responsible for pollution and to identify minor spills before they can cause widespread damage. The detection of pollutants using GPR [1], was based on the research of Basson [2]. Basson [3] presented a combination of GPR and FDEM methods to detect and monitor saline contaminants in agricultural fields. Goldshleger [4, 5] demonstrated the ability to detect saline-affected soils using remote-sensing methods, toward improved management of these soils. Basson [6] described the detection of subsurface water/sewage/drainage pipe systems and leaks/contamination from such pipes. Ben-Dor [7, 8] reviewed remote-sensing–based methods to assess soil salinity and improve the management of salinity-affected soils. Ly and Chui [9] developed accurate representations of weep holes and leaky sewage pipes, and further showed the systems\' long-term and short-term responses to rainfall events. Their simulation results provided a better understanding of local-scale migration of sewage leaks from a sewage pipe to nearby storm water drains. The last few years in Israel have seen increasing use of new methods based on active remote-sensing tools to study subsoil quality. These tools include GPR and underground monitoring systems measuring spatial moisture content, such as FDEM in the subsurface. The use of GPR is based on a method that was originally developed for measuring sand dunes of medium moisture content at an unsaturated resolution of a few percentage points [2]. The GPR helped define the possible reason for emerging high-salinity areas, such as a subsurface regional structure that reduces water infiltration into the deeper groundwater position [5]. The FDEM method provided a very important view of salt contamination in the soil layers (except the root zone layer) and also pinpointed areas with salinity problems. The images obtained from FDEM readings provided a subsurface view that also helped identify the reason for the high salinity in certain areas. In the soil salinity experiment in Israel, a severe defect in the drainage pipelines could be observed, which helped the farmers solve the problem before the subsequent season [5].
The present study focuses on the development of these electromagnetic (EM) methods to replace conventional acoustic methods for the identification of sewage pipe leaks. EM methods provide an additional advantage in that they allow mapping the fluid transport system in the subsurface. Leak-detection systems using GPR and FDEM are not limited to large amounts of water, but can also detect leaks of tens of liters per hour, because they can locate increases in pipes’ or tanks’ environmental moisture content that amount to only a few percentage points. The importance and uniqueness of this research lies in the development of practical tools to provide a snapshot of the spatial changes in soil moisture content to depths of about 3–4 m (in areas with asphalt overlay) at relatively low cost, in real time or close to real time. Spatial measurements performed using GPR and FDEM systems allow monitoring many tens of thousands of measurement points per hectare, thus providing a picture of the spatial situation along the pipelines. The main purpose of this study was to develop a method for detecting sewage leaks using the above-proposed geophysical methods, as the resultant contaminants can severely affect public health. We focused on identifying, locating and characterizing such leaks in sewage pipes in residential and industrial areas.
In recent years, there has been an increase in the use of active remote-sensing tools, such as GPR (Figure 1a) and subsurface FDEM (Figure 1b), for measuring the subsurface\'s EM velocity and dielectric constant (GPR), and its electrical conductivity profile and magnetic susceptibility (FDEM).
Taking measurements with the RAMAC GPR (a) and Gem-2 FDEM (b) in the study area.
Passive remote-sensing spectroscopy of ground surface and cross-sections using an optical fiber termed SPSP (subsurface-penetrating spectral probe), developed [10] and have been conducted as well. This study focuses on remote-sensing tools to replace acoustic methods [11, 12, 13]. EM methods provide the added advantage of being able to map underground liquid-carrying pipelines. Ground leak-detection systems using GPR and FDEM are not limited to large amounts of water: small leaks of tens of liters per hour can be detected in the environment by comparing medium-dry to minimum moisture content in the pipeline and the canal zone.
Our aim was to develop practical tools that would provide a snapshot of changes in spatial soil moisture content to depths of about 3–4 m in areas covered with asphalt at relatively low cost and in real time. The spatial measurements were performed with FDEM and GPR systems that allow measuring tens of thousands of points per hectare and thus enable monitoring the spatial situation along the pipeline.
Traditionally, the electrical method “measures” apparent resistivity using electrodes that require ground contact in a DC electrical survey, while the EM method “measures” apparent conductivity without ground contact. The EM method, known as a “potential method”, involves transmitting and receiving EM fields, commonly using a set of coils. The common unit of resistivity is ohm-m and conductivity is its inverse, in Siemen/m. The apparent resistivity
where
Similarly, apparent conductivity is only same as the true conductivity when the earth is a homogeneous half space. As an example, consider a pair of horizontal coils separated by a distance
where
Geometry of the horizontal coplanar electromagnetic sensor over layered earth where
The in-phase and quadrate responses as a function of induction number (from Huang and Won, 2003).
Figure 3 shows the responses of the Gem-2 sensor over a half space as a function of induction number:
where
The depth of investigation of an EM system can be estimated using the skin depth
The skin depth and the ability to transmit in several frequencies allows us to perform “frequency sounding” using a multifrequency sensor, thereby resolving different depths of penetration as sketched in Figure 4.
Frequency sounding for various depths using a multifrequency FDEM sensor such as Gem-2.
GPR, a reflection-scattering imaging method, is widely used for subsurface imaging in geophysics. GPR uses high frequencies (wavelengths; MHz–GHz). EM waves may form images of the subsurface by transmitting radar pulses into the ground and receiving the deflected waves from the interfaces below. Using wave methods and analysis, GPR images can be analyzed for their derived electrical properties and subsurface characteristics and for spatial mapping of water content [2, 3]. The range resolution is a function of the subsurface dielectric constants and the wave\'s frequency. It may vary from several centimeters to several tens of centimeters at the relevant effective frequencies [19, 20] For a certain wavelength, the penetration of GPR waves into the subsurface is mainly a function of the host material\'s conductivity, and therefore GPR waves decay significantly in conductive and saline soils. Using wave methods and analysis, GPR images can be analyzed for their derived electrical properties and subsurface characteristics and for spatial mapping of water content [2,3], as described in the following model.
The connection between the EM velocity and dielectric constant is expressed as:
where
The dielectric constant of water (
The difference in the effective dielectric constant of "dry" and "wet" soils is mainly a function of the ratio between the air and water volumes, when the volumes are normalized to:
\n\t\t\t\tthen:
\n\t\t\t\tThe maximal soil–water absorbency is a strong function of the effective porosity.
Ariel is a small city (about 20,000 residents) in Israel, located in the central highland region known as the Samarian Hills. It is situated 40 km (25 miles) east of Tel Aviv and 40 km west of the Jordan River. It is situated 700 m (more than 2000 feet) above sea level. The city stretches over 12 km (8 miles) in length and 2 km in width. The research was performed with Yuvalim, the company that is responsible for maintaining the water and sewage network in the Ariel area and for supplying available water to residents. The mutual research was performed to identify sewage leaks before they pollute and damage the surrounding area. The research was supported by the Israeli Water Authority. The work was performed in several stages.
Areas were selected in Ariel for system calibration (Figure 5). Two areas were chosen for the method calibration: the first was an industrial area and the second a residential area, both with well-mapped networks of water and sewage pipes. These areas were selected on the basis of information from computerized data, observations, field visits, use of orthophotos, aerial photography and geological and pedological data.
Maps showing Ariel\'s location (a) and the drainage infrastructure, sewerage and water supply for this city (b).
To characterize the pedological structure of the subsurface layers, excavations were performed. We sampled grain size, void content and porosity, moisture content, soil density and soil characteristics. We dug a channel in an underground sewage pipe replacement area at the experimental sites. Figure 6 presents the characterization of the sub layer.
Soil subsurface cross-section at site 1. Wooden pegs mark the changing soil layers.
The soil in the area is red Mediterranean, also known as Terra Rossa [21] and Lithic ruptic Xerochrept [22]. Terra Rossa occurs in areas where heavy rainfall dissolves carbon from the parent calcium carbonate rock and silicates are leached out of the soil, leaving residual deposits that are rich in iron hydroxides, causing the red color. Such areas are usually depressions within limestone. The soil was sampled in a 0.5-m-wide ditch at a depth of 2 m. The area has an easterly aspect, with an average elevation of 400 m above sea level. The local slopes vary between 7% and 25%. Soil texture was clay loam with an average composition of 45% sand, 25% silt and 30% clay. The sand content increased toward the lower part of the area. The average lime content was 30%. Rock fragments of up to 40 cm appeared together with the soil.
Calibration of the GPR system to the subsurface properties of the cross-section in a dry state (without leakage) is shown in Figure 7. The depth to the pipe was measured in a nearby manhole.
Part of the GPR profile performed for calibration of the GPR system in the Ariel industrial zone, on the road close to a rubber factory. The black circle displays diffraction created by the drain pipe. Above it, the trench is detected as well. The horizontal scale describes the measurement location (in meters) along the profile. The vertical scales describe the time (in nanoseconds) and depth (in meters). The amplitude–intensity scale is shown as well.
Figure 7 shows the results of advanced processing of a cross-section for calibration of the system in the industrial area. On the horizontal scale, simulations are described above the measurement location along the incision in meters; the vertical scales describe the time and depth of the reflections on a timescale of 50 ns and scale depth of 2.5 m below the surface (the strength of the reflections is graded according to the color scale in Figure 7, where the diffraction created by the drainage pipe can be deduced from a return time from the pipe of approximately 32 ns). The diffraction depth is 2.45 m, and the data from the system matches the data measured on the ground. This adaptation makes it possible to determine the velocity of the EM wave. The average measured subsurface speed of the EM wave (
The experimental site for sewage pipeline and manhole leaks was located near Ariel\'s old stadium, not far from HaAtsmaut Street (Figure 8, blue rectangle), where a project for the replacement of old sewer pipes has been initiated.
The experimental site is located at the western end of the sewage line adjacent to HaAtsmaut Street (blue rectangle). It includes 12-in. diameter iron pipes carrying on the order of 1000–1200 m3 sewage water per day, and an average 100 m3/h during peak flow.
Leakage was initiated in two places at the western site by cracking the sewer pipes close to their bottom side. One crack was made about 6 m from the sewage pit in the northern iron pipe using an electrical disk that created a wedge-shaped hole 15–20 cm in diameter; the second crack was also a circle of 15–20 cm diameter in the lower part of the pipe (Figure 9). The experimental site was monitored daily by radar and FDEM before the start of and during the controlled leakage.
Pictures of the two cracks made in the sewer pipes for the controlled leakage experiment.
Daily monitoring with the FDEM method included five cross-sections: four were parallel to the sewer pipeline and the fifth was above it, running on each side of the pipeline at a distance of 0.5 m. During the experiment, FDEM scanning was performed to qualify the effect of moisture on the soil cross-section. Figure 10 shows the status of the subsurface before the start of the controlled leak; it was in a relatively dry state characteristic of the month of May at this site.
Map of the integrated electrical conductivity at 60,025 Hz before the start of the controlled leak at the western site (locations of the measurements are shown by the blue rectangle in
Figure 11 shows a pronounced increase in electrical conductivity of about 40 mS/m after 4 days of controlled leakage. The area has high conductivity because of changes in wetness due to a significant increase in liquid as a result of the sewage flow.
Map of the electrical conductivity at 60,025 Hz after about 4 days of leakage. Measurements were collected during the sewage leak, under wet conditions, with the GEM-2 sensor (locations of the measurements are shown by the blue rectangle in
The results of the FDEM measurements conducted 10 days after the beginning of the controlled leak are presented in Figure 12. This picture may look similar to Figure 11 in terms of colors, but their intensity has increased due to an increase in the conductivity values to about 152 mS/m.
Map of integrated electrical conductivity at 60,025 Hz. Measurements were collected with the FDEM system, under wet conditions, after 10 days of controlled leakage (locations of the measurements are shown by the blue rectangle in
On the map in Figure 12, low visibility, reflecting low electrical conductivity, is shown in blue-green shades, high visibility in red-colored shades. Purple indicates sewer leakage on the background of the driest area, highlighting the differences in moisture. A wide area can be seen west of the pipe (black line in Figure 12) with relatively low electrical conductivity compared to the rest of the region. Northeast of the pipe, there is high electrical conductivity resulting from the spillover of sewage water.
Figure 13 shows maps made by FDEM monitoring of electrical conductivity at various frequencies in the first tested area. The maps are arranged, from left to right, at increasing frequencies and depth: the frequencies were 2,025 Hz, 4,725 Hz, 11,025 Hz, 25,725 Hz and 60,025 Hz, each frequency representing a 30 cm increase in depth. The low-visibility electrical conductivity is represented by blue-green hues, and the high-visibility electrical conductivity by red-purple hues. There were a few quantitative differences in the map scales.
Maps made by FDEM monitoring of electrical conductivity at 2,025 Hz, 4,725 Hz, 11,025 Hz, 25,725 Hz and 60,025 Hz. The lower EC values are represented by blue-green hues, and the higher EC values by red-purple hues. There were a few quantitative differences between the maps\' scales.
Four sections, two on each side of the sewer, were monitored by GPR and are shown in Figure 14. The distance between the main radar cross-sectional cuts was approximately 0.5 m. The radar sections shown in Figure 14 were collected with an antenna at a nominal frequency of 250 MHz over the location of the underground sewage pipe at the first (western) test site. The first cross-section was obtained before the leak started and reflects the typical dry state of the ground in May. An incision was made a few days after the initiation of the leak and shows a relatively wet subsoil. The right cross-section shows an incision made at a lower depth, 10 days after leak initiation, indicating a further increase in wetness. Similar data processing was carried out for the three cross-sections to highlight their differences.
Soil moisture reflected by GPR cross-section (locations of the measurements are shown by the blue rectangle in
Moisture content was computed on the basis of subsurface GPR and FDEM measurements and its spatial spread was obtained for calibration and wetness testing with water- and sewage-carrying pipelines. In these experiments, radar velocities were measured and dielectric constants were computed. Their correlations were used to measure the moisture content from data collected in the residential and industrial neighborhoods.
The computation of moisture content using GPR was based on the method developed by Basson [2] From the calibration measurements conducted at the end of May 2012, the average subsurface EM wave velocity was 0.093 ± 0.001 m/ns. The calculated dielectric constant during this period was about 10.4. This value is low but not minimal, as minimal moisture content is typically found in the mid-to-late summer months (according to data from the Israel Meteorological Service, the rain that accumulated in the area in the months before the GPR measurements amounted to about 161 mm).
The velocity of EM waves in a substance is mainly a function of that substance\'s bulk dielectric properties and moisture content. When a substance is composed of a mixture of materials, the velocity is a function of their mixing ratios. In the case of a subsurface environment, we can treat the substance as a bulk property composed of soil, rock, minerals and organic materials mixed with air and water. When the rate of air increases, the velocity increases as well. However, when the moisture content increases, the average dielectric constant decreases as well and fro equation (6) it can be seen that the EM velocity (v) decreases as well.
The difference in the effective dielectric constant of "dry" and "wet" soils is mainly a function of the ratio between the air and water volumes, when the volumes are normalized according to equations (7) and (8). The maximal soil–water absorbency is a strong function of the effective porosity. For soils in the Ariel region, the effective porosity can vary from 40% to 60%. We used an average effective porosity of 50% in our computations. Therefore, the possible mixing ratios relative to the normalized volume are:
\n\t\t\tSince
The radar wave velocity for “dry” soil at the surface will be measured and is expected to vary with the GPR and its value,
We develop a moisture content model using relative values of the moisture content (based on Equations (6–14)) causing an increase in electrical conductivity as measured by the FDEM. We had to consider the overall subsurface features, such as texture, density and effective porosity, as well as the content of salts in soils irrigated with brackish effluent water. The model results are presented in the graph in Figure 15.
Volumetric moisture content calculated from measurements and from the FDEM model in the experimental zones in Ariel (accuracy ±10% of the measured value).
We introduced a combination of GPR and FDEM orthogonal methods to detect subsurface leaks from a sewage pipeline system. The rationale for this combination is to increase the probability of detection, especially in complex urban environments and when the soil–rock setting can vary from relatively resistive to relatively conductive. The results of our study indicate that even minor leaks, such as the minor controlled leaks created in the experiment, and changes in the subsurface moisture content can be accurately detected. We could detect sewage leakage, as well as its progress. The combination of the two methods enabled not only the detection of the leak but also a qualitative assessment of its size. Factors affecting the ability to detect leaks were limited by the soil–rock conductivity, as well as the density of the terrain and subterrain systems and structures. The geophysical methods may detect sewage effluent flow paths as well as the contaminant in the soil.
The limestone and dolomite bedrock in the Ariel area is suitable for GPR mapping. The clarity of the GPR profile enabled analysis and interpretation of the physical data with good accuracy. We could detect sewage leakage, as well as its progress. The anomalous moisture of the leakage accumulating around the sewage pit in the southwest research area validated the efficiency of the methods.
The research was supported by the Israel water authority and by the Water Cooperation of Yuvalim. We would like to thank Mr. Omer Shamir from GeoSense for assistance with the data collection.
The design of scaffold materials that can guide tissue regeneration is a very challenging goal [1]. In addition, to support and promote the growth and differentiation of specific cells, an ideal scaffold requires careful control of the material’s structure in the range of nanometers to centimeters, and some natural materials with complex structure exist in nature, which provides ideas for the design of ideal scaffolds [2]. These natural materials, such as mammal bones, abalone pearl layers and fish scales, which are composed of multi-layer biominerals and biopolymers, have complex microstructure, which can control the crack growth and fracture in three-dimensional (3D) direction, producing much more strength and toughness than their constituent materials [3, 4, 5]. Jellyfish and sea anemones, with a water content of up to 90%, show that their gelatinous bodies exhibit exciting mechanical properties and are able to respond quickly to various environmental stimuli [6, 7, 8]. There are also some soft support tissues (such as tendons, ligaments, meniscus, and cartilage), showing softness, toughness and impact resistance [9]. Because of the beneficial properties of natural composite materials, the design of bionic materials has attracted significant attention. Bio-inspired material is considered as a kind of material inspired by nature or biology and then developed by simulating some characteristics [10], and usually, the bio-inspired materials provide better functions than synthetic materials [11].
\nHowever, there are still many limitations on the fabrication of bio-inspired materials using traditional material manufacturing technology because they cannot accurately control the distribution and spatial trend of micro-holes inside the materials, and it is challenging to produce the contour matching with natural materials [12, 13]. Recently, 3D bioprinting technology has become a promising tool for manufacturing materials with high-precision, which can overcome the limitations compare with the traditional methods, and finally can eventually produce complex and delicate biomimetic 3D structures. Also, 3D bioprinting technology realizes the automatic biological preparation of cell-laden structure through the layered deposition of bio-inks
In recent years, in tissue engineering development, many materials have been developed to meet the needs of 3D bioprinting. The most common 3D bioprinting materials are metals, engineering plastics, photosensitive resins, bioplastics and polymer hydrogels. The bio-inspired hydrogels are very similar to natural extracellular matrix (ECM) and display potential advantages in tissue engineering [19]. Bio-inspired hydrogel provides an adequate and porous microenvironment that allows good nutrition and oxygen to diffuse into the encapsulated cells and can be modified to guide cellular processes with various physical, chemical, and biological cues [20]. Besides, these hydrogels are usually non-toxic or low toxic and have good reproducibility. Next, the 3D bioprinting techniques used for the fabrication of bio-inspired hydrogels were summarized, and the materials used for 3D bioprinting were outlined. This chapter also focuses on the applications of bio-inspired hydrogels.
\nThere are several available 3D bioprinting techniques for fabricating bio-inspired hydrogels, including inkjet bioprinting, laser-assisted bioprinting, extrusion bioprinting, and stereolithography, as shown in \nFigure 1\n [21].
\nBioprinting techniques mainly include inkjet, laser-assisted, extrusion and stereolithography [
During the inkjet bioprinting process, biomaterials are selectively placed on the construction platform layer by layer until the required structure is formed. The first inkjet printers for bioprinting applications were improved versions of commercial two-dimensional ink printers [22]. For the inkjet bioprinting, the ink in the ink cartridge is replaced by biomaterials, and the paper is replaced by an electronically controlled lifting table to provide the control of the third dimension Z-axis in addition to the X-and Y-axes. The bioprinter based on inkjet printing technology is customized to process and print biomaterials with higher resolution, accuracy and speed [16]. Inkjet bioprinters use thermal or acoustic forces to spray droplets onto the substrate, which can support or form part of the final structure [23]. Thermal inkjet uses a heating element to induce the evaporation of a small volume of bioink in a reservoir, thereby forming and ejecting a small droplet. Therefore, in the printing process, this method keeps the cells at high temperature (300°C) for several microseconds (about 2 microseconds), which may lead to the formation of transient pores in the cell membrane [16]. Using the thermal inkjet printer, Solis et al., studied the effect of heat generated by the thermal ink-jet bio printer and found that the survival rate of Chinese hamster ovary (CHO) cells was 89% [24]. Such survival rate of cells could be greatly improved by using a piezoelectric inkjet printer, the generation and injection of droplets are realized by applying external voltage to control the mechanical deformation of piezoelectric transducer, which prevents the temperature from rising to the super physiological level [25]. Compaan et al. used alginate as the sacrificial material to prepare cell-supported silk fibroin hydrogels with a clear structure based on the piezoelectric inkjet 3D bioprinting system. The printed tubular structure has a diameter of 5 mm, a height of 2.5 or 5.0 mm and a thickness of about 400 microns. Moreover, the effect of citrate treatment on the printing was compared. The results showed that alginate removal and alginate removal could enable cells to extend and contact each other and form a cell network in the whole hydrogel [26].
\nThe advantages of inkjet bioprinting mainly include: low cost due to its similar structure to commercial printers, high printing speed due to the ability of the print head to support parallel operation mode, and relatively high unit survival rate (usually from 80–90%) determined by many experimental results. However, the risks of cells and materials exposed to thermal and mechanical stresses, low droplet directionality, uneven droplet size, frequent nozzle plugging, and unreliable cell encapsulation have brought considerable limitations to the application in tissue engineering [27].
\nThe typical laser-assisted biological printing device include pulsed laser beams, focusing systems, and donor bands that respond to laser stimuli, consisting of glass covered with laser energy absorbing layers, and biomaterial layers (such as cells/hydrogel composite) prepared in liquid and receiving substrates for ribbons. The principle of laser-assisted bioprinting is to apply high-energy pulse laser (usually near-infrared laser) to the donor color band coated with bioink. This laser pulse evaporates a part of the donor layer, forms a high-pressure bubble on the interface of the bioink layer, and pushes the materials containing cells to the receiving substrate [16, 25, 28]. Compared with inkjet bioprinting, laser-assisted bioprinting can avoid the problem of jamming cell or material, also can avoid direct contact with the printer and biological ink at the same time. The non-contact biological printing method can choose much more types of ink, resulting in printing materials with wider range of viscosity [28].
\nThe laser pulse energy, ECM thickness, and bioink viscosity can influence cell viability. The higher the laser energy is, the higher the cell death rate is, but the increase of membrane thickness and bioink viscosity will lead to an increase of cell viability. Guillotin et al. studied the effects of bioink viscosity, laser energy and printing speed on printing resolution. The microscale resolution and 5 kHz printing speed could be achieved, and the laser-assisted bioprinting could combine cells with ECM to produce soft tissue with high cell density
The extrusion bioprinting can fabricate 3D cell carriers for tissue regeneration. The prepolymer solutions need to be prepared first, and almost all types of prepolymer solutions with different viscosities and aggregates with high cell density can be printed with extruded bioprinters [28]. Different from printing small droplets onto the platform, the extrusion bioprinting continuously deposit hydrogel filaments within a diameter of 150–300 microns to generate 3D structures. Common extrusion bioprinting method includes pneumatic, piston-driven, and screw-driven dispensing. In pneumatic dispensing, air pressure provides the required driving force, while in piston and screw-driven dispensing, vertical and rotating mechanical forces start printing respectively [30]. There are three main factors that decide the printability of extrusion bioprinting, mainly including the adjustability of viscosity, the bioink phase before extrusion, and the material-specific bio-manufacturing window [31]. Extrusion bioprinters have been used to produce various tissue types, such as aortic valves, branching vascular trees, in vitro drug movement and tumor models [32]. Although the manufacturing time may be prolonged for high-resolution complex structures, the structures have been manufactured from the clinically related tissue size to the microtissue in the microfluidic chamber. Furthermore, it is convenient to combine cells with bioactive agents, because that the heating process is not involved [33]. Compared with inkjet 3D bioprinting, extrusion bioprinters can achieve a continuous flow of biomaterials, thus achieving the simplicity of operation and a broader selection of biomaterials, including polymers, acellular matrices, cellular hydrogels, spheres and aggregates [34].
\nAmong all the bioprinting technologies, stereolithography (SLA) 3D bioprinting display much more advantages over extrusion or ink-jet bioprinting technology [28]. SLA is based on the polymerization of photosensitive polymers, and the digital mirror array controls the light band in the projection field to achieve selective crosslinking of each layer of the hydrogel prepolymer solution [35]. No matter how intricate a layer’s pattern is, the printing time is the same because the whole pattern is projected on the printing plane. Therefore, the printer only needs a movable table in the vertical direction, which significantly simplifies the control of the printer. The cell encapsulated scaffold fabricated by the SLA system can achieve 100 μm resolution with printing time less than 1 hour, also maintain very high cell viability (90%) [36]. The above properties make SLA practical for fabricating delicate construct for tissue engineering. Arcaute et al. used composite lithography technology and two different molecular weight of polyethylene glycol (PEG) to prepare composite multilayer 3D structure of PEG hydrogel, and the properties of prepared hydrogel were influenced by photo-initiator and photosensitive polymer concentration. Besides, the prepared PEG hydrogel supports attachment, proliferation and differentiation of bovine chondrocytes, providing evidence for the applicability of resins for cartilage tissue engineering [37]. Valentin et al. prepared the sodium alginate precursor solution based on ion crosslinking, and different concentrations of cationic sources, such as barium carbonate, magnesium carbonate and calcium carbonate, and photo acid generator (PAG), diphenyliodonium nitrate were used, and the sodium alginate hydrogel was printed by SLA. The printed alginate hydrogel exhibited different mechanical and physical properties when crosslinked with two kinds of cations. The microstructures with variable height could be printed with optimized precursor formulations. Due to the high resolution, the 3D fabrication of natural and synthetic polyelectrolyte hydrogels via SLA enables lab-on-a-chip devices, soft sensors and actuators, and other biologically-inspired devices [38].
\nHydrogels are considered as the gold standard materials for 3D bioprinting because they can provide a flexible and hydrated cross-linked network, similar to the natural extracellular matrix, in which cells can survive [39]. The polymers prepared for hydrogels can be classified into natural and synthetic polymers [40]. The natural polymers include alginate, chitosan, hyaluronic acid, gelatin, and so on, and the synthetic polymers mainly include polyacrylamide (PAAm), polyvinyl alcohol (PVA), polyethylene glycol (PEG), polylactic acid (PLA), and so on [41, 42].
\nMost hydrogels prepared by natural polymers have the advantages of good hydrophilicity, good biocompatibility, specific enzymatic degradation, and contain various active functional groups and structural domains, and display better interaction with cells to promote cell proliferation and differentiation.
\nAlginate is extracted from alginate plants, is a kind of natural high molecular, composing of β-d-mannuronate (M) and α-l-guluronate (G). Alginate has been widely used in tissue engineering because of its advantages of abundant production, low price, good biocompatibility, and abundant functional groups, which are suitable for the preparation of bioink for 3D bioprinting [43, 44]. Alginate can react with CaCO3 to release bivalent Ca2+ and then form an ionic crosslinking hydrogel bonded with -COO- on G unit of alginate G unit, to achieve the controllability of alginate ion crosslinking. The alginate hydrogel has high toughness and good mechanical properties, but the degradation rate of the alginate hydrogel is not controllable [45].
\nChitosan is the product of deacetylation of chitin, which has a straight-chain structure and positive charge due to the presence of amino groups. Because of the useful biological function and biocompatibility, the degradation by microorganisms, chitosan has been widely concerned and applied in various industries [46]. The chitosan ink can be directly printed in air, and then the chitosan scaffold is refined by physical gelation. A chitosan hydrogel that satisfies both biocompatibility and mechanical properties has been obtained, and it has been confirmed that chitosan hydrogel can guide cell growth [47].
\n(A) Fabrication of rapid gelation and tough GelMA/HA-NB/LAP hydrogel for DLP-based printing. (B) the skin analogous with sophisticated two-layer gel structure was fabricated via 3D bioprinting. (a) the bioink was printed with a layer-by-layer style using a DLP-based 3D printer. (b, c) the structure of native skin was displayed in CAD images. (d) the lower layer view of the scaffold was shown. (e) CAD images of different designed microchannel size and the printed products. (f) the elastic compressibility of products. (g) Compressive Young’s modulus [
Gelatin is the hydrolysate of collagen, which contains many arginine-glycine-aspartic-acid (RGD) sequences and matrix metalloproteinase (MMP) target sequences, which enhance cell adhesion and cellular microenvironment remodeling respectively [49]. Because of biodegradability, biocompatibility, and low antigenicity, gelatin is attractive for bio-inspired hydrogel [50]. Lewis et al. used gelatin as a bioink to print into a specific 3D geometry using 3D bioprinting, which can regulate the biological processes of hepatocytes, enhance protein function, and facilitate cell proliferation and differentiation [51]. Another commonly used gelatin derivative is to acylate gelatin to form gelatin methacrylamide (GelMA) [52]. Zhou et al. used GelMA, N-(2-aminoethyl)-4-(4-(hydroxymethyl)-2-methoxy-5-nitrosophenoxy) butanamide linked hyaluronic acid (HA-NB) and photo-initiator lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP) as biomimetic bioink to fabricated a bio-inspired 3D tissue construct via the digital light process (DLP)-based 3D bioprinting technology for ski regeneration (\nFigure 2\n) [48]. Bhise et al. used GelMA to carry out Hep G2/C3A cells to prepare biomimetic 3D liver structure hydrogel through bioprinting technology. A bionic human body chip of liver tissue was prepared by bioreactor. The toxicity response test of this chip in the test of acetaminophen is similar to that reported
Hyaluronic acid (HA) is a kind of biocompatible non-sulfated glycosaminoglycan composed of N-acetylglucosamine and D-glucuronic acid repeated disaccharide units [54]. It is abundant in tissues including cartilage, neurons and skin. HA is of intrinsic biological importance because it binds to receptors such as CD44, can be degraded by oxidative species and hyaluronidase, and is related to the function and structure of development, wound healing and adult tissues. Because of biocompatibility, biodegradability, and natural biological function, HA hydrogels are widely used in various application fields [55]. Besides, the HA hydrogel can energize cell viability and promote osteoblasts to differentiate into cartilage. Unlike collagen and other proteins, the sequence of HA is different from species and its antigenicity is low, so it is especially promising as an injectable hydrogel.
\nSeveral other natural polymers, such as collagen, agarose, carrageenan, fibrin, heparin, chondroitin sulfate, cellulose, hemicellulose, lignin, and so on, could be used for hydrogels using 3D bioprinting [21]. However, natural hydrogels lack adequate mechanical properties, especially when implanted
The hydrogels fabricated using synthetic polymers have the advantages of long service life, strong water absorption, and high gel strength [41]. Polyacrylamide (PA) is a general designation of acrylamide homopolymer and copolymer. PA is a kind of water-soluble polymer, which has many amide groups in its structure and is easy to form hydrogen bond, so it has good stability and flocculation and is easy to be chemically modified. Ahn et al. grafted poly (N-isopropylacrylamide) (PNIPAAm) onto the framework of sodium alginate and synthesized sodium alginate PNIPAAm polymer micelles by self-assembly in aqueous solution, and the micelles could be used for the encapsulation of anticancer drug adriamycin [56]. Polyethylene glycol (PEG) is another synthetic polymer, and it has no toxicity and irritation, has good biocompatibility, and can be discharged from the body through the kidney. It has been widely used in the field of biomedicine [57]. Gao et al. constructed the polyethylene glycol diacrylate (PEGDA) hydrogel with uniform distribution of human mesenchymal stem cells (hMSCs) inside by simultaneous photopolymerization with commercial thermojet printers. hMSCs filled in 3D PEGDA hydrogel showed no deposition during culture and showed a chondrogenic phenotype [58]. Wang et al. prepared an injectable hydrogel through
Polylactic acid (PLA) is a kind of polymer, which is made of lactic acid as the primary raw material, and through polymerization, in which the performance can be adjusted by the structure [60]. Senatov et al. prepared PLA/hyaluronic acid (HA) interconnected porous scaffold via a melt-wire method; the 3D printing technique avoided thermal degradation of PLA, the porosity and pore size of the scaffold could be well controlled. The porous PLA/HA scaffold with 15% HA has a considerable crack resistance and can work for a long time under the stress of 21 MPa, which was potential for bone tissue engineering applications [61].
\nPolyvinyl alcohol (PVA) is a synthetic water-soluble polymer, it has good biodegradation, biocompatibility, and no side effects on the human body [62]. PVA has been widely used in ophthalmology, wound dressing, artificial joint, and so on [42, 63]. Shi et al. prepared an injectable dynamic hydrogel using HA grafted with PVA and phenyl boric acid (PBA). The synthesized HA-PBA-PVA dynamic hydrogel has the reactive oxygen species reactivity and the scavenging activity of active oxygen. Furthermore, the hydrogel had good biocompatibility to the encapsulated neural precursor cells (NPC), and its ability to scavenge reactive oxygen species could protect the NPC cells from the damage of reactive oxygen species. The HA-PBA-PVA hydrogel could be used as bioink for 3D biological printing to prepare multilayer and cell loaded structures. The NPC cells showed good viability (85 ± 2% of living cells) after extrusion and maintained the excellent viability of 81 ± 2% of living cells after 3 days of culture. The results indicated that multifunctional injectable and ROS responsive self-healing HA-PBA-PVA dynamic hydrogels were expected to be candidates for 3D culture and 3D bioprinting [64].
\nBesides, there are also many other synthetic polymers for the fabrication of bio-inspired hydrogels, such as Pluronic and derivatives, PEG or polyethylene oxide (PEO) based block copolymers, poly(L-glutamic acid), poly(propylene fumarate), methoxy polyethylene glycol, and so on. Though, the synthetic polymers can precisely control their gel structure and properties and have better physical and chemical stability and more raw materials to prepare bio-inspired hydrogels. However, it is necessary to pay attention to the possible biocompatibility of unreacted monomers and residual initiators during the preparation of synthetic polymer materials, and the biocompatibility could be greatly improved via compositing or liking with natural polymers [65, 66, 67].
\nTissue regeneration research is aim to develop substitute for damaged or diseased tissues or organs using principles of life science, engineering and medicine synergistically. It is crucial to fabricate the substitute as scaffolds, which is inspired by the natural 3D structure of tissue. The natural ECM regulates essential cellular functions, such as adhesion, migration, proliferation, differentiation and morphogenesis [68]. It is important of mimicking the ECM with dynamic nature using 3D bioprinting techniques, and the bio-inspired hydrogels via such techniques displayed potential applications in tissue regeneration, such as cartilage tissue, vascularized engineered tissue, bone tissue, skin regeneration, heart tissue, aortic valve conduits, muscle-tendon, and so on [69]. For example, Alexander et al. displayed a chemically and mechanically biomimetic filler-free bioink for 3D bioprinting of soft neural tissues, as shown in \nFigure 3\n. The thiolated Pluronic F-127, dopamine-conjugated (DC) gelatin, and DC hyaluronic acid were used as bioinks via a thiol-catechol reaction and photocuring; the storage modulus of the cured bioinks ranged from 6.7 to 11.7 kPa. The micro-extrusion 3D bioprinting was used to fabricate free-standing cell-laden tissue constructs. The Rodent Schwann cells, rodent neuronal cells, and human glioma cell-laden tissue constructs were printed and cultured over seven days and exhibited excellent viability, which has implications in micro physiological neural systems for neural tissue regenerative medicine [70]. Several works could be found in a recent study that focuses on the specific properties of bio-inspired hydrogels for tissue regeneration, such as high strength structures [30]. Also, the enhancement of printing resolution and versatility is vital for tissue regeneration. For example, the self-healing hydrogels were used to support the direct 3D bioprinting with high resolution by utilizing shear-thinning hydrogels, then the constructs could be printed in any direction [71]. The bio-inspired hydrogels could be accomplished via
(A) Native ECM components of neural tissue were combined with a synthetic polymer for microextrusion 3D bioprinting of soft, free-standing neural tissues. (B) Two curing pathways, including UV light exposure, and chelation of dopamine groups with iron (III), are shown to the formulation of photocuring containing methacrylated dopamine-conjugated gelatin. With the increase of PF127-SH content, the compressive properties of inks cured through UV exposure or chelation increased.
The bio-inspired hydrogels via 3D bioprinting can be applied for wound dressing and wearable devices, which are considered as important applications, especially in recent years. Skin plays an essential role in protecting the body from external damages, such as abrasions, lacerations, and burns, and so on. The full-thickness defects of the dermis layers are the most challenging wounds to heal because of the limitation of self-repairing capability; thus, the skin regeneration of skin with skin appendages still remains a tough challenge [72]. 3D bioprinting is being applied to fabricate skin constructs using biomaterial scaffolds with or without cells, to address the need for skin tissues suitable for transplantation for wound healing therapy. The natural polymers, including cellulose, collagen and chitin, alginate, and hyaluronic acids are employed to synthesis skin constructs due to the favorable biocompatibility, biodegradation, low-toxicity or nontoxicity, high moisture content, high availability and mechanical stability [73]. Feifei et al. fabricated gelatin methacrylate (GelMA) based bioink to print functional living skin using DLP-based 3D printing (\nFigure 2\n), while the printed skin could promote skin regeneration and neovascularization via mimicking the physiological structure of natural skin [48].
\nFurthermore, the bio-inspired hydrogels could not only be functionalized on skin regeneration but also as medical wearable devices. The conductive hydrogels could be designed and fabricated to acquire electronic devices with conductive, capacitive, switching properties, image displaying, and motion sensing [74]. Meihong et al. developed conductive, healable, and self-adhesive hybrid network hydrogels based on conductive functionalized single-wall carbon nanotube (FSWCNT), PVA and polydopamine. The prepared hydrogel exhibits fast self-healing ability around 2 s, high self-healing efficiency of about 99%, and robust adhesiveness, which could be used for healable, adhesive, and soft human-motion sensors [75]. Zijian et al. synthesized a stretchable, self-healing and conductive hydrogel based on gelatin-enhanced hydrophobic association poly(acrylamide-
The bio-inspired hydrogels could also be used in drug delivery system, such as protein carriers, anti-inflammatory drug carriers, in the pharmaceutical industry [79]. Rana et al. designed a magnetic natural hydrogel based on alginate, gelatin, and iron oxide magnetic nanoparticles as an efficient drug delivery system, the drug doxorubicin hydrochloride (DOX) was loaded, the anticancer activity against Hela cells could be regulated by the release of DOX from hydrogels [80]. Maling et al. provided a proof-of-concept of detoxification using a 3D-printed biomimetic nanocomposite construct in the hydrogel. A bio-inspired 3D detoxification device by installing polydiacetylene (PDA) nanoparticles in a 3D matrix was fabricated using dynamic optical projection stereolithography (DOPsL) technology; the nanoparticles could attract, capture and sense toxins, while the 3D matrix with a modified liver lobule microstructure allows toxins to be trapped efficiently [36]. The bio-inspired hydrogels via multi-materials 3D bioprinting can easy regulate the loading and release profiles of drugs, which show potentials as biomedicines.
\nThe design paradigms shift from 2D to 3D has revolutionized the way of bio-inspired hydrogels for materials components, engineered constructs,
The future outlook of 3D bioprinting for fabrication of bio-inspired tissues for tissue engineering applications [
The 3D bioprinting has changed the way bio-inspired hydrogels fabricated, and expanded the applications of bio-inspired hydrogels, including tissue regeneration, wound dressing, wearable devices, and pharmaceutical applications, and so on. In this chapter, the available 3D bioprinting techniques were described, the advantages and disadvantages of each printing technology were outlined. Then, the natural and synthetic polymers used for the fabrication of bio-inspired hydrogels via 3D bioprinting were introduced. The applications of bio-inspired hydrogels were focused. At last, the future outlook of bio-inspired hydrogels for tissue engineering were summarized. The bio-inspired hydrogels produced from 3D bioprinting still lacking sufficient clinical evidence, as more clinical trials evaluating bio-inspired hydrogels are still required.
\nThis research was funded by the National Natural Science Foundation of China No. 31700840.
\nThe authors declare no conflict of interest.
ADSCs | adipose derived stem cells |
CAD | \n |
CAM | computer-aided manufacturing |
CHO | Chinese hamster ovary |
DLP | digital light process |
DOPsL | dynamic optical projection stereolithography |
DOX | doxorubicin hydrochloride |
ECM | extracellular matrix |
FSWCNT | functionalized single-wall carbon nanotube |
GelMA | gelatin methacrylamide |
HA | hyaluronic acid |
HA-SH | thiol hyaluronic acid |
hMSCs | human mesenchymal stem cells |
LAP | lithium phenyl-2,4,6-trimethylbenzoylphosphinate |
MMP | matrix metalloproteinase |
MRI | magnetic resonance imaging |
NB | N-(2-aminoethyl)-4-(4-(hydroxymethyl)-2-methoxy-5-nitrosophenoxy) butanamide |
NPC | neural precursor cells |
PA | polyacrylamide |
PAAm | polyacrylamide |
PAG | photo acid generator |
PBA | phenyl boric acid |
PDA | polydiacetylene |
PEG | polyethylene glycol |
PEGDA | polyethylene glycol diacrylate |
PEO | polyethylene oxide |
PLA | polylactic acid |
PNIPAAm | poly (N-isopropylacrylamide) |
PVA | polyvinyl alcohol |
RGD | arginine-glycine-aspartic-acid |
SLA | stereolithography |
TA-PEGDA | tetraniline polyethylene glycol diacrylate |
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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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