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Hydrogel patches with high toughness, stretchability, and adhesive properties are critical to healthcare applications including wound dressings and wearable devices. Gelatin methacryloyl (GelMA) provides a highly biocompatible and accessible hydrogel platform. However, low tissue adhesion and poor mechanical properties of cross-linked GelMA patches ( i.e ., brittleness and low stretchability) have been major obstacles to their application for sealing and repair of wounds. Here, we show that adding dopamine (DA) moieties in larger quantities than those of conjugated counterparts to the GelMA prepolymer solution followed by alkaline DA oxidation could result in robust mechanical and adhesive properties in GelMA-based hydrogels. In this way, cross-linked patches with ∼140% stretchability and ∼19 000 J/m 3 toughness, which correspond to ∼5.7 and ∼3.3× improvement, respectively, compared to that of GelMA controls, were obtained. The DA oxidization in the prepolymer solution was found to play an important role in activating adhesive properties of cross-linked GelMA patches (∼4.0 and ∼6.9× increase in adhesion force under tensile and shear modes, respectively) due to the presence of reactive oxidized quinone species. We further conducted a parametric study on the factors such as UV light parameters, the photoinitiator type ( i.e ., lithium phenyl-2,4,6-trimethylbenzoylphosphinate, LAP, versus 2-hydroxy-4'-(2-hydroxyethoxy)-2-methylpropiophenone, Irgacure 2959), and alkaline DA oxidation to tune the cross-linking density and thereby hydrogel compliance for better adhesive properties. The superior adhesion performance of the resulting hydrogel along with in vitro cytocompatibility demonstrated its potential for use in skin-attachable substrates.

Tissue adhesives have been developed for sealing tissue damaged in surgery. Among these, sheet-type adhesives require a relatively long time to adhere to biological tissue under wet conditions. To address this clinical problem, we fabricated a tissue-adhesive fiber sheet (AdFS) based on decanyl group (C10) modified Alaska pollock-derived gelatin (C10-ApGltn) using electrospinning. Ultraviolet (UV) irradiation of the AdFS was performed to increase the affinity between the AdFS and wet biological tissue by introducing hydrophilic functional groups. The UV irradiated AdFS (UV-C10-AdFS) strongly adhered to porcine pleura within 2 min under wet conditions and showed higher burst strength compared with the original ApGltn (Org-ApGltn) sheet. Hematoxylin-eosin stained sections revealed that a dense UV-C10-AdFS layer remained on the surface of the porcine pleura even after burst strength measurement. Moreover, UV-C10-AdFS has excellent cytocompatibility and efficiently supports the growth of L929 cells. UV-C10-AdFS is a promising adhesive material for sealing wet biological tissue., Competing Interests: Declaration of competing interest The authors have no competing interests to declare., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Bioinks play a key role in determining the capability of the biofabricatoin processes and the resolution of the printed constructs. Excellent biocompatibility, tunable physical properties, and ease of chemical or biological modifications of gelatin methacryloyl (GelMA) have made it an attractive choice as bioinks for biomanufacturing of various tissues or organs. However, the current preparation methods for GelMA-based bioinks lack the ability to tailor their physical properties for desired bioprinting methods. Inherently, GelMA prepolymer solution exhibits a fast sol-gel transition at room temperature, which is a hurdle for its use in stereolithography (SLA) bioprinting. Here, synthesis parameters are optimized such as solvents, pH, and reaction time to develop GelMA bioinks which have a slow sol-gel transition at room temperature and visible light crosslinkable functions. A total of eight GelMA combinations are identified as suitable for digital light processing (DLP)-based SLA (DLP-SLA) bioprinting through systematic characterizations of their physical and rheological properties. Out of various types of GelMA, those synthesized in reverse osmosis (RO) purified water (referred to as RO-GelMA) are regarded as most suitable to achieve high DLP-SLA printing resolution. RO-GelMA-based bioinks are also found to be biocompatible showing high survival rates of encapsulated cells in the photocrosslinked gels. Additionally, the astrocytes and fibroblasts are observed to grow and integrate well within the bioprinted constructs. The bioink's superior physical and photocrosslinking properties offer pathways of tuning the scaffold microenvironment and highlight the applicability of developed GelMA bioinks in various tissue engineering and regenerative medicine applications., (© 2020 Wiley-VCH GmbH.)

Adipose tissue engineering aims to provide solutions to patients who require tissue reconstruction following mastectomies or other soft tissue trauma. Mesenchymal stromal cells (MSCs) robustly differentiate into the adipogenic lineage and are attractive candidates for adipose tissue engineering. This work investigates whether pore size modulates adipogenic differentiation of MSCs toward identifying optimal scaffold pore size and whether pore size modulates spatial infiltration of adipogenically differentiated cells. To assess this, extrusion-based 3D printing is used to fabricate photo-crosslinkable gelatin-based scaffolds with pore sizes in the range of 200-600 µm. The adipogenic differentiation of MSCs seeded onto these scaffolds is evaluated and robust lipid droplet formation is observed across all scaffold groups as early as after day 6 of culture. Expression of adipogenic genes on scaffolds increases significantly over time, compared to TCP controls. Furthermore, it is found that the spatial distribution of cells is dependent on the scaffold pore size, with larger pores leading to a more uniform spatial distribution of adipogenically differentiated cells. Overall, these data provide first insights into the role of scaffold pore size on MSC-based adipogenic differentiation and contribute toward the rational design of biomaterials for adipose tissue engineering in 3D volumetric spaces., (© 2020 The Authors. Published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

The effects of ultrasound-assisted extraction (UAE) and microwave-assisted extraction (MAE) methods on molecular and physicochemical characteristics of the resultant gelatin were examined. Before extraction procedure, we investigated the optimum pH for swelling of Common carp by-products, which is an important pretreatment for gelatin production. The highest swelling yield was achieved at pH 13 among pH 1-14 with unit intervals. Results indicated that the UAE gelatin has a higher gel strength, viscosity, melting point, and gelling point. The power and time of sonication showed a reverse relation with these characteristics. In addition, as the time of microwave heating was raised, the gel strength, viscosity, melting point, and gelling point were decreased. The FT-IR spectra showed similar peaks but the Amide B in UAE gelatin slightly vanished. The electrophoretic pattern also revealed the higher gel strength and viscosity of UAE gelatin due to the higher intensity of α and β chains compared to MAE gelatin. It can be concluded from all of the results of this study that the produced gelatin using these procedures can be a good source of gelatin in food and drug industries., (© 2019 Wiley Periodicals, Inc.)

In this study, the composite of carboxymethyl cellulose (CMC) film modified with gelatin and TiO 2 -Ag nanoparticles (CMC/Gel/TiO 2 -Ag) was prepared and some properties of synthesized film including physicochemical and photocatalytic properties were investigated. FT-IR results showed that new interactions between the film components were created. Scanning electron microscopy (SEM) results showed that the TiO 2 -Ag particles with 50-100 nm distributed in the CMC/Gelatin film. The results of the mechanical test showed that the TiO 2 -Ag nanoparticles at low concentrations increased tensile strength (TS) and decreased strain to break (STB), but with increasing nanoparticles concentrations, TS decreased and STB increased. Photocatalytic study showed that the prepared CMC/Gel/TiO 2 -Ag film has good photocatalytic property. Gas chromatography was used to study photocatalytic effects of film. Increasing TiO 2 -Ag nano particles on the film increases the photocatalytic activity of films against NH3, ethanol and benzene., (Copyright © 2019. Published by Elsevier Ltd.)

In this study, the cyto-compatibility and cellular functionality of cell-laden gelatin-methacryloyl (Gel-MA) hydrogels fabricated using a set of photo-initiators which absorb in 400-450 nm of the visible light range are investigated. Gel-MA hydrogels cross-linked using ruthenium (Ru) and sodium persulfate (SPS), are characterized to have comparable physico-mechanical properties as Gel-MA gels photo-polymerized using more conventionally adopted photo-initiators, such as 1-[4-(2-hydroxyethoxy)-phenyl]-2-hydroxy-2-methyl-1-propan-1-one (Irgacure 2959) and lithium phenyl(2,4,6-trimethylbenzoyl) phosphinate (LAP). It is demonstrated that the Ru/SPS system has a less adverse effect on the viability and metabolic activity of human articular chondrocytes encapsulated in Gel-MA hydrogels for up to 35 days. Furthermore, cell-laden constructs cross-linked using the Ru/SPS system have significantly higher glycosaminoglycan content and re-differentiation capacity as compared to cells encapsulated using I2959 and LAP. Moreover, the Ru/SPS system offers significantly greater light penetration depth as compared to the I2959 system, allowing thick (10 mm) Gel-MA hydrogels to be fabricated with homogenous cross-linking density throughout the construct. These results demonstrate the considerable advantages of the Ru/SPS system over traditional UV polymerizing systems in terms of clinical relevance and practicability for applications such as cell encapsulation, biofabrication, and in situ cross-linking of injectable hydrogels., (© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Hard capsules are made from gelatin, an organic polymer obtained through the hydrolysis of collagen present in animal tissues. Gelatin can be degraded by microorganisms and some strategies can be used to control contaminating micro-organisms. Gamma irradiation is considered as an effective sterilization method; however, its application can alter the chemical structure of the irradiated product. Samples of hard gelatin capsules were irradiated at doses of 5, 15, and 25 kGy at room temperature. The characterizations of the physical and chemical effects were evaluated by scanning electron microscopy, Fourier-transform infrared spectroscopy, X-ray diffractometry, and differential scanning calorimetry techniques. Furthermore, hard gelatin capsule samples were dissolved and inoculated with Bacillus subtilis, a Gram-positive spore-forming bacterium, to evaluate the effect of gamma ray radiation on bacterial counts. The results showed that gamma radiation did not interfere on physical parameters of the capsule, such as moisture content, mass, body and cap length, and disintegration time. Nevertheless, differential scanning calorimetry results demonstrated changes in the glass transition temperature, indicating the formation of crosslinking in irradiated capsules. It was observed that there were significant reductions on the inoculated bacterial population starting from the lowest irradiation dose and there was no detection of bacterial growth from the 15 kGy dose, while in the non-irradiated samples were found with 10 4  CFU mL -1 of bacteria. Therefore, this work concludes that the gamma radiation is effective on the reduction of the microbial population, cause discrete physical-chemical alterations, and could be used as a hard capsule sterilization technique.

This study investigated a novel method of photosensitizer-induced cross-linking (using riboflavin as a sensitizer) to improve the structural and physicochemical properties of gelatin-based edible films with different glycerol concentrations (25% and 50%) during different UV exposure times (2, 4 and 6 h). The films' tensile strength was enhanced significantly for both glycerol concentrations with increasing UV exposure times compared to the control film, so that the highest tensile strength was observed for films with 25% glycerol and 6 h of UV exposure (25%-6 h). The films' tensile strength declined and the elongation at break increased about three times when the glycerol concentration was increased to 50% with 6 h exposure. The photosensitizer-induced cross-linking significantly reduced the films' solubility and permeability. The UV-treated films exhibited very good barrier properties against UV, with zero light transmission at a wavelength of 200 to 350 nm. Moreover, no toxicity was found in any of the films. In addition, Fourier transform infrared spectroscopy and differential scanning calorimetry findings revealed a good interaction between functional groups of riboflavin (as the sensitizer) and gelatin in the 25%-6 h film. Therefore, this new method can be a suitable alternative to chemical methods of cross-linking biopolymers., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Background: In this paper, two kinds of commonly used vegetable carbon black (VCB, 3000 mesh; nano) at 50 g kg -1 concentration (based on dried gelatin) were added to 48 g kg -1 of fish gelatin (GEL) solutions and their effects on mechanical, barrier and anti-photooxidation properties of GEL films were investigated., Results: From the SEM images, it was shown that compared with 3000 mesh VCB (1-2 μm), nano VCB (100-200 nm) made the microstructure of GEL film more compact and more gelatin chains were cross-linked by nano VCB. The addition of nano VCB significantly increased gelatin film strength with the greatest tensile strength of 52.589 MPa and stiffness with the highest Young's modulus of 968.874 MPa, but led to the reduction of film elongation. Also, the VCB presence significantly improved water vapour and oxygen barrier properties of GEL film. Importantly, nano VCB increased GEL film with better UV barrier property due to its stronger UV absorption nature when compared with micron VCB. This property could help in the preservation of oil samples in the photooxidation accelerated test., Conclusion: With improved properties, the nano VCB-reinforced GEL film may have great potential for application in the edible packaging field, especially for the anti-photooxidation property. © 2017 Society of Chemical Industry., (© 2017 Society of Chemical Industry.)

Recent developments in radiation therapy aimed at more precise dose delivery along with higher dose gradients (dose painting) and more efficient dose delivery with higher dose rates e.g. flattening filter free (FFF) irradiation. Magnetic-resonance-imaging based polymer gel dosimetry offers 3D information for precise dose delivery techniques. Many of the proposed polymer gels have been reported to exhibit a dose response, measured as relaxation rate ΔR2 (D) , which is dose rate dependent. A lack of or a reduced dose-rate sensitivity is very important for dosimetric accuracy, especially with regard to the increasing clinical use of FFF irradiation protocols with LINACs at high dose rates. Some commonly used polymer gels are based on Methacrylic-Acid-Gel-Initiated-by-Copper (MAGIC). Here, we report on the dose sensitivity (ΔR2/ΔD) of MAGIC-type gels with different oxygen scavenger concentration for their specific dependence on the applied dose rate in order to improve the dosimetric performance, especially for high dose rates. A preclinical x-ray machine ('Yxlon', E  =  200 kV) was used for irradiation to cover a range of dose rates from low [Formula: see text] min   =  0.6 Gy min -1 to high [Formula: see text] max   =  18 Gy min -1 . The dose response was evaluated using R2-imaging of the gel on a human high-field (7T) MR-scanner. The results indicate that all of the investigated dose rates had an impact on the dose response in polymer gel dosimeters, being strongest in the high dose region and less effective for low dose levels. The absolute dose rate dependence [Formula: see text] of the dose response in MAGIC-type gel is significantly reduced using higher concentrations of oxygen scavenger at the expense of reduced dose sensitivity. For quantitative dose evaluations the relative dose rate dependence of a polymer gel, normalized to its sensitivity is important. Based on this normalized sensitivity the dose rate sensitivity was reduced distinctly using an increased oxygen scavenger concentration with reference to standard MAGIC-type gel formulation at high dose rate levels. The proposed gel composition with high oxygen scavenger concentration exhibits a larger linear active dose response and might be used especially in FFF-radiation applications and preclinical dosimetry at high dose rates. We propose in general to use high dose rates for calibration and evaluation as the change in relative dose sensitivity is reduced at higher dose rates in all of the investigated gel types.

The aim of this study was to investigate the efficacy of power ultrasound without using any heating stage in solubilizeing gelatin dispersions, and to characterize the mechanical and microstructural properties of the resulting gels using texture analysis and scanning electron microscopy, respectively. Usually to prepare a gel from gelatin, a primary heating stage of at about 40C or above is required to solubilize gelatin macromolecules. In this study solubilizing gelatin dispersions using power ultrasound without any heating was successfully performed. For solubilising gelatin, an ultrasound equipment with a frequency of 20 kHz, amplitude of 100% and power range of 50-150 W was used. Aqueous gelatin dispersions (4% w/v) were subjected to ultrasound for different times (40-240 s) at a constant temperature of 13C. Applying ultrasound to gelatin dispersions caused increases in water absorption and water solubility of the hydrocolloid. The textural parameters of the resulting gelatin gels, increased with increasing time and power of ultrasound. Moreover, a generalized Maxwell model with three elements was used for calculating relaxation times of the gels. The microstructural observations by SEM showed that the structural cohesiveness of the gels increased by increasing ultrasonication time. Ultrasound-assisted solubilization of gelatin can have emerging implications for industrial uses in pharmaceuticals, food and non-food systems., Practical Applications: Usually to prepare a gel from gelatin, a primary heating stage of at about 40C or above is required to solubilize gelatin macromolecules. Therefore, the use of gelatin as a hydrocolloid in food processings or pharmaceutical formulations which lack a heating step has been a technological and practical challenge. In this study solubilizing gelatin dispersions using power ultrasound without any heating was successfully performed. Ultrasound-assisted solubilisation of gelatin can have emerging implications for industrial uses in pharmaceuticals, food, and non-food systems, for example, to conserve heat sensitive compounds., (© 2016 Wiley Periodicals, Inc.)

The objective of this work was to display the effect of electron beam accelerator doses on properties of plasticized fish gelatin film. Electron spin resonance indicates free radical formation during irradiation, which might induce intermolecular cross-linking. Tensile strength for gelatin film significantly increases after irradiation (improved by 30% for 60 kGy). The vapour permeability is weakly affected by irradiation. Surface tension and its polar component increase significantly and are in accordance with the increase of wettability. So, irradiation may change the orientation of polar groups of gelatin at the film surface and crosslink the hydrophobic amino acids. No modification of the crystallinity of the film is observed. These findings suggest that if structure changes, it only occurs in the amorphous phase of the gelatin matrix. It is also observed that irradiation enhances the thermal stability of the gelatin film, by increasing the glass transition temperature and the degradation temperature., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Biofunctional scaffolds that support the adhesion, proliferation, and osteo-differentiation of mesenchymal stem cells (MSCs) are critical for bone tissue engineering. In this study, a simple in situ UV-crosslinking strategy was utilized to fabricate gelatin electrospun fibrous (GEF) scaffolds with multiple biosignals, including cell adhesive Arg-Gly-Asp (RGD) peptide, osteo-conductive hydroxyapatite (HAp) nanoparticles, and osteo-inductive bone morphogenic protein-2 (BMP-2). The adhesion and proliferation of MSCs on the GEF scaffolds were improved by the incorporation of RGD. Meanwhile, the incorporation of HAp and BMP-2 enhanced osteo-differentiation of MSCs. The three incorporated bio-factors exert a synergistic effect on osteogenesis of MSCs in the GEF scaffolds. This strategy of incorporating multiple biomolecules could be used to fabricate crosslinked electrospun scaffolds of natural polymers for tissue-engineering applications., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The first derivatives of gelatin and type I collagen fluorescence spectra were characterized in order to describe the effect of free radicals on pyridinoline (PYD) cross-links. The different gas saturation conditions were used to investigate the effect of different free radicals. An analysis of first derivative fluorescence spectra suggests that PYD cross-link fluorescence emission is composed of three peaks in gelatin, but only two in type I collagen. The PYD cross-link was photo-degraded more than other gases in the presence of O2. This suggests that the singlet oxygen ((1)O2) plays a key role when using photodynamic therapy to treat skin fibrosis disease with Hypocrellin B (HB).

In vitro engineering of autologous fatty tissue constructs is still a major challenge for the treatment of congenital deformities, tumor resections or high-graded burns. In this study, we evaluated the suitability of photo-crosslinkable methacrylated gelatin (GM) and mature adipocytes as components for the composition of three-dimensional fatty tissue constructs. Cytocompatibility evaluations of the GM and the photoinitiator Lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP) showed no cytotoxicity in the relevant range of concentrations. Matrix stiffness of cell-laden hydrogels was adjusted to native fatty tissue by tuning the degree of crosslinking and was shown to be comparable to that of native fatty tissue. Mature adipocytes were then cultured for 14 days within the GM resulting in a fatty tissue construct loaded with viable cells expressing cell markers perilipin A and laminin. This work demonstrates that mature adipocytes are a highly valuable cell source for the composition of fatty tissue equivalents in vitro. Photo-crosslinkable methacrylated gelatin is an excellent tissue scaffold and a promising bioink for new printing techniques due to its biocompatibility and tunable properties., (© The Author(s) 2015.)

Photocrosslinkable hydrogels are frequently used in cartilage tissue engineering, with crosslinking systems relying on cytotoxic photoinitiators and ultraviolet (UV) light to form permanent hydrogels. These systems are rarely assessed in terms of optimization of photoinitiator or UV dosage, with non-cytotoxic concentrations from literature deemed sufficient. We hypothesized that the number of reactive functional groups present within a hydrogel polymer is highly relevant when crosslinking, affording cytoprotection to chondrocytes by preferentially interacting with the highly reactive radicals that are formed during UV-mediated activation of a photoinitiator. This was tested using two photocrosslinkable hydrogel systems: gelatin methacrylamide (GelMA) and gellan gum methacrylate (GGMA). We further assessed the effects of two different UV dosages on chondrocyte differentiation while subject to a single photoinitiator dosage in the GGMA system. Most notably, we found that a higher ratio of reactive groups to photoinitiator molecules offers cytoprotective effects, and future developments in photocrosslinkable hydrogel technology may involve assessment of such ratios. In contrast, we found there to be no effect of UV on chondrocyte differentiation at the two chosen dosages. Overall the optimization of photocrosslinkable systems is of great value in cartilage tissue engineering and these data provide a groundwork for such concepts to be developed further., Statement of Significance: Photocrosslinkable hydrogels, which use photoinitiators and predominantly ultraviolet light to form stable matrices for cell encapsulation and tissue development, are promising for cartilage tissue engineering. While both photoinitiators and ultraviolet light can damage cells, these systems have generally not been optimized. We propose that the ratio of reactive functional groups within a polymer to photoinitiator molecules is a critical parameter for optimization of photocrosslinkable hydrogels. Using photocrosslinkable gelatin and gellan gum, we found that a higher ratio of reactive groups to photoinitiator molecules protected chondrocytes, but did not affect chondrocyte differentiation. The principle of cytoprotection by functional groups developed in this work will be of great value in optimizing photocrosslinkable hydrogel systems for cartilage and other tissue engineering applications., (Copyright © 2015 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

Tilapia skin gelatin, pig skin gelatin, and their mousse premixes were exposed to UV irradiation for 103, 206, and 309 kJ/cm(2). All samples after 309 kJ/cm(2) exposure exhibited a significant increase in gel strength, gel forming ability as well as viscosity of solutions. It was shown that UV treatment could also improve the pig skin gelatin foam stability and foam formation ability compared to those of tilapia skin gelatin. Nevertheless, the panelists gave the lowest scores to mousse made with 309 kJ/cm(2) UV-irradiated premix mousse pig skin gelatin. Tilapia skin gelatin could be used as a substitute ingredient for premix mousse made from pig skin gelatin., (© The Author(s) 2014 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.)

Modular tissue engineering holds great potential in regenerating natural complex tissues by engineering three-dimensional modular scaffolds with predefined geometry and biological characters. In modular tissue-like construction, a scaffold with an appropriate mechanical rigidity for assembling fabrication and high biocompatibility for cell survival is the key to the successful bioconstruction. In this work, a series of composite hydrogels (GH0, GH1, GH2, and GH3) based on a combination of methacrylated gelatin (GelMA) and hydroxyapatite (HA) was exploited to enhance hydrogel mechanical rigidity and promote cell functional expression for osteon biofabrication. These composite hydrogels presented a lower swelling ratio, higher mechanical moduli, and better biocompatibility when compared to the pure GelMA hydrogel. Furthermore, on the basis of the composite hydrogel and photolithograph technology, we successfully constructed an osteon-like concentric double-ring structure in which the inner ring encapsulating human umbilical vascular endothelial cells (HUVECs) was designed to imitate blood vessel tubule while the outer ring encapsulating human osteoblast-like cells (MG63s) acts as part of bone. During the coculture period, MG63s and HUVECs exhibited not only satisfying growth status but also the enhanced genic expression of osteogenesis-related and angiogenesis-related differentiations. These results demonstrate this GelMA-HA composite hydrogel system is promising for modular tissue engineering.

A layered gelatin-chitosan hydrogel with graded composition was prepared via photocrosslinking to simulate the polysaccharide/collagen composition of the natural tissue and mimic the multi-layered gradient structure of the cartilage-bone interface tissue. Firstly, gelatin and carboxymethyl chitosan were reacted with glycidyl methacrylate (GMA) to obtain methacrylated gelatin (Gtn-GMA) and carboxymethyl chitosan (CS-GMA). Then, the mixed solutions of Gtn-GMA in different methacrylation degrees with CS-GMA were prepared to form the superficial, transitional and deep layers of the hydrogel, respectively under the irradiation of ultraviolet light, while polyhedral oligomeric silsesquioxane was introduced in the deep layer to improve the mechanical properties. Results suggested that the pore sizes of the superficial, transitional and deep layers of the layered hydrogel were 115 ± 30, 94 ± 34, 51 ± 12 μm, respectively and their porosities were all higher than 80 %. The compressive strengths of them were 165 ± 54, 565 ± 50 and 993 ± 108 kPa, respectively and the strain of the gradient hydrogel decreased along the thickness direction, similar to the natural tissue. The in vitro cytotoxicity results showed that the hydrogel had good cytocompatibility and the in vivo repair results of osteochondral defect demonstrated remarkable recovery by using the gradient gelatin-chitosan hydrogel, especially when the hydrogel loading transforming growth factor-β1. Therefore, it was suggested that the prepared layered gelatin-chitosan hydrogel in this study could be potentially used to promote cartilage-bone interface tissue repair.

Sterilization through γ-irradiation has been reported to affect collagen mechanical properties, but its possible effects on gelatin based materials have not been investigated up to now. Herein we report the results of a mechanical, chemical and thermal study performed on gelatin films before and after γ-irradiation. The investigation was performed on uncrosslinked films as well as on crosslinked films. To this aim, two common crosslinking agents, glutaraldehyde and genipin, at different concentration (0.15, 0.30 and 0.67%) were used. The results indicate that sterilization significantly affects the mechanical properties of uncrosslinked films, whereas it displays a modest effect on gelatin swelling, release in solution, thermal stability and molecular structure. Both glutaraldehyde and genipin enhance the mechanical properties and stability in solution of the gelatin films. In particular, the values of Young modulus increase as a function of crosslinker concentration up to about 10 and 18 MPa for genipin and glutaraldehyde treated samples respectively. The results of in vitro study demonstrate that the films crosslinked with genipin do not display any cytotoxic reaction, whereas glutaraldehyde crosslinking provokes an acute and dose dependent cytotoxic effect.

Chondroprogenitor cells encapsulated in a chondrogenically supportive, three-dimensional hydrogel scaffold represents a promising, regenerative approach to articular cartilage repair. In this study, we have developed an injectable, biodegradable methacrylated gelatin (mGL)-based hydrogel capable of rapid gelation via visible light (VL)-activated crosslinking in air or aqueous solution. The mild photocrosslinking conditions permitted the incorporation of cells during the gelation process. Encapsulated human-bone-marrow-derived mesenchymal stem cells (hBMSCs) showed high, long-term viability (up to 90 days) throughout the scaffold. To assess the applicability of the mGL hydrogel for cartilage tissue engineering, we have evaluated the efficacy of chondrogenesis of the encapsulated hBMSCs, using hBMSCs seeded in agarose as control. The ability of hBMSC-laden mGL constructs to integrate with host tissues after implantation was further investigated utilizing an in vitro cartilage repair model. The results showed that the mGL hydrogel, which could be photopolymerized in air and aqueous solution, supports hBMSC growth and TGF-β3-induced chondrogenesis. Compared with agarose, mGL constructs laden with hBMSCs are mechanically stronger with time, and integrate well with native cartilage tissue upon implantation based on push-out mechanical testing. VL-photocrosslinked mGL scaffold thus represents a promising scaffold for cell-based repair and resurfacing of articular cartilage defects.

A major goal in the application of hydrogels for tissue engineering scaffolds, especially for load-bearing tissues such as cartilage, is to develop hydrogels with high mechanical strength. In this study, a double-network (DN) strategy was used to engineer strong hydrogels that can encapsulate cells. We improved upon previously studied double-network (DN) hydrogels by using a processing condition compatible with cell survival. The DN hydrogels were created by a two-step photocrosslinking using gellan gum methacrylate (GGMA) for the rigid and brittle first network, and gelatin methacrylamide (GelMA) for the soft and ductile second network. We controlled the degree of methacrylation of each polymer so that they obtain relevant mechanical properties as each network. The DN was formed by photocrosslinking the GGMA, diffusing GelMA into the first network, and photocrosslinking the GelMA to form the second network. The formation of the DN was examined by diffusion tests of the large GelMA molecules into the GGMA network, the resulting enhancement in the mechanical properties, and the difference in mechanical properties between GGMA/GelMA single networks (SN) and DNs. The resulting DN hydrogels exhibited the compressive failure stress of up to 6.9 MPa, which approaches the strength of cartilage. It was found that there is an optimal range of the crosslink density of the second network for high strength of DN hydrogels. DN hydrogels with a higher mass ratio of GelMA to GGMA exhibited higher strength, which shows promise in developing even stronger DN hydrogels in the future. Three dimensional (3D) encapsulation of NIH-3T3 fibroblasts and the following viability test showed the cell-compatibility of the DN formation process. Given the high strength and the ability to encapsulate cells, the DN hydrogels made from photocrosslinkable macromolecules could be useful for the regeneration of load-bearing tissues., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Hydrogels are three-dimensional water-insoluble hydrophilic natural or synthetic polymer networks made up of crosslinked water-soluble polymers. The purpose of this study was to develop and directly compare photo crosslinked hydrogels on the basis of pure gelatin, alginate and hyaluronic acid as well as their blends. The functionalization of starting materials with methacrylate moieties was evaluated by 1H-NMR spectroscopy. Hydrogels were prepared from methacrylates by photo cross-linking using UV light. The effect of changing the hydrogel composition was quantified through examination of hydrogel swelling behavior and rheological properties. In addition, the viability and adhesion of neonatal rat cardiomyocytes (NRCM) seeded onto the hydrogels was examined by in vivo imaging of NRCM-mediated scaffold contraction as well as by histological evaluation after immunostaining. Biological testing showed good biocompatibility and cell survival in the presence of all materials discussed. Adhesion of cells could only be observed in the presence of gelatin. Blends of gelatin, alginate and hyaluronic acid are promising candidates for the generation of non-toxic, biocompatible hydrogel scaffolds for tissue engineering. Variation of individual compound ratios in the blends can be used for a precise control of mechanical properties and may allow wide-ranging uses in various tissue engineering applications with different mechanical requirements.

A novel visible light-crosslinkable porcine gelatin was prepared for gelation and micropatterning. The preparation employed a photo-oxidation-induced crosslinking mechanism. First, furfuryl groups were incorporated into the gelatin. Second, the modified gelatin was mixed in water with Rose Bengal, which is a visible light sensitizer. Irradiation by visible light solidified the aqueous solution. In addition, when the solution was cast on a plate, dried and photo-irradiated in the presence of a photomask a micropattern was formed that matched the micropattern on the photomask. The gelatin-immobilized regions enhanced cell adhesion. It was also confirmed that the gelatin incorporating furfuryl and Rose Bengal have no significant toxicity. The photo-crosslinkable gelatin was employed as a direct pulp capping material in the dental field. Considering these results, this system could be useful as a new type of visible light-induced crosslinkable biosealant., (2010 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

Bioprinting by the codeposition of cells and biomaterials is constrained by the availability of printable materials. Herein we describe a novel macromonomer, a new two-step photocrosslinking strategy, and the use of a simple rapid prototyping system to print a proof-of-concept tubular construct. First, we synthesized the methacrylated ethanolamide derivative of gelatin (GE-MA). Second, partial photochemical cocrosslinking of GE-MA with methacrylated hyaluronic acid (HA-MA) gave an extrudable gel-like fluid. Third, the new HA-MA:GE-MA hydrogels were biocompatible, supporting cell attachment and proliferation of HepG2 C3A, Int-407, and NIH 3T3 cells in vitro. Moreover, hydrogels injected subcutaneously in nude mice produced no inflammatory response. Fourth, using the Fab@Home printing system, we printed a tubular tissue construct. The partially crosslinked hydrogels were extruded from a syringe into a designed base layer, and irradiated again to create a firmer structure. The computer-driven protocol was iterated to complete a cellularized tubular construct with a cell-free core and a cell-free structural halo. Cells encapsulated within this printed construct were viable in culture, and gradually remodeled the synthetic extracellular matrix environment to a naturally secreted extracellular matrix. This two-step photocrosslinkable biomaterial addresses an unmet need for printable hydrogels useful in tissue engineering.

Purpose: Polymer-based gel dosimeter (MAGIC type) is a preferable phantom material for PET range verification of proton beam therapy. However, improvement in elemental tissue equivalency (specifically O/C ratio) is very desirable to ensure realistic time-activity measurements., Methods: Glucose and urea was added to the original MAGIC formulation to adjust the O/C ratio. The dose responses of the new formulations were tested with MRI transverse relaxation rate (R2) measurements., Results: The new ingredients improved not only the elemental composition but also the sensitivity of the MAGIC gel. The O/C ratios of our new gels agree with that of soft tissue within 1%. The slopes of dose response curves were 1.6-2.7 times larger with glucose. The melting point also increased by 5 degrees C. Further addition of urea resulted in a similar slope but with an increased intercept and a decreased melting point., Conclusions: Our improved MAGIC gel formulations have higher sensitivity and better elemental tissue equivalency for 3D dosimetry applications involving nuclear reactions.

We investigated the amount and distribution of hydrogen peroxide (H(2)O(2)) generated in a solid gelatin sample irradiated by heavy ion (carbon) beam. We irradiated the gelatin sample, which contained a nitroxyl radical (4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl, TEMPOL), with a 290-MeV/nucleon carbon beam (~128 Gy). To verify the distribution of H(2)O(2) generation in the irradiated sample, we employed both electron paramagnetic resonance (EPR) spectroscopic and magnetic resonance (MR) imaging methods based on H(2)O(2)-dependent paramagnetic loss of TEMPOL. We obtained a distribution profile of the H(2)O(2)-dependent reaction in the gelatin sample when we irradiated gelatin samples with carbon beams with several different linear energy transfer (LET) values. Because the profiles of oxygen consumption in the gelatin sample measured by L-band EPR oxymetry and of the H(2)O(2)-dependent reaction have almost the same shape, the profile of the H(2)O(2)-dependent reaction can be used as an estimation of the profile of the generation of H(2)O(2). The H(2)O(2) profile in one intact gelatin sample scanned by 7-tesla MR imaging showed a similar shape as a result of the EPR experiment. We obtained several hundreds of micromolars of H(2)O(2) generated in a gelatin sample irradiated by carbon beam when 200 Gy was given at the surface of the sample. H(2)O(2) distribution was almost flat, with only a slight peak just before the end of the beam.

The development of novel three-dimensional cell culture platforms for the culture of aortic valvular interstitial cells (VICs) has been fraught with many challenges. Although the most tunable, purely synthetic systems have not been successful at promoting cell survivability or function. On the other hand, entirely natural materials lack mechanical integrity. Here we explore a novel hybrid system consisting of gelatin macromers synthetically modified with methacrylate functionalities allowing for photoencapsulation of cells. Scanning electron microscopy observations show a microporous structure induced during polymerization within the hydrogel. This porous structure was tunable with polymerization rate and did not appear to have interconnected pores. Treatment with collagenase caused bulk erosion indicating enzymatic degradation controls the matrix remodeling. VICs, an important cell line for heart valve tissue engineering, were photoencapsulated and examined for cell-directed migration and differentiation. VICs were able to achieve their native morphology within 2 weeks of culture. The addition of the pro-fibrotic growth factor, transforming growth factor-beta1, accelerated this process and also was capable of inducing enhanced alpha-smooth muscle actin and collagen-1 expression, indicating a differentiation from quiescent fibroblasts to active myofibroblasts as demonstrated by quantitative real-time polymerase chain reaction and immunohistochemistry. Although these studies were limited to VICs, this novel hydrogel system may also be useful for studying other fibroblastic cell types.

The 2H NMR spectrum of HDO in gelatin gel, that is stretched inside silicone rubber tubing, displays a well resolved doublet. Spectra were obtained with a range of offset frequencies of partially saturating radio-frequency (RF) radiation. The resulting steady-state irradiation envelope (also referred to as a 'z-spectrum') has the peculiar feature that maximal suppression of the doublet occurs when the irradiation is applied exactly at the centre frequency, between the two HDO peaks. We present a quantum mechanical explanation for this phenomenon. It is shown that the phenomenon is the result of double quantum transitions. The analysis is extendable to more complex quadrupolar and dipolar-coupled systems of other nuclides. It has implications for enhancement of contrast in magnetic resonance imaging of heterogeneous systems using dipolar and quadrupolar interactions.

Lasing requires an active gain medium and a feedback mechanism. In conventional lasers the feedback is provided externally, e.g. by mirrors. An alternate approach is through Bloch waves in photonic crystals composed of periodic dielectric materials in which propagation of light in certain frequency ranges, known as photonic bandgaps, is forbidden. Compared to periodic crystals, quasicrystals have higher symmetry and are more favorable for the formation of photonic bandgaps. Hence quasicrystals are more efficient in providing the feedback mechanism for lasing. Here we report observation of lasing at visible wavelengths from dye-doped three-dimensional icosahedral quasicrystals fabricated in dichromate gelatin emulsions using a novel seven-beam optical interference holographic method. Multi-directional lasing exhibiting the icosahedral symmetry was observed. The lasing modes and pattern were explained by using the lasing condition expressed in the reciprocal lattice space of the icosahedral quasicrystal.

Radiation-sensitive polymer gels for clinical dosimetry have been intensively investigated with magnetic resonance imaging (MRI) because the transversal magnetic relaxation time is dependent on irradiation dose. MRI is expensive and not easily available in most clinics. For this reason, low-cost, quick and easy-to-use potential alternatives such as optical computed tomography (CT), x-ray CT or ultrasound attenuation CT have also been studied by others. Here, we instead evaluate the dose dependence of the elastic material property, Young's modulus and the dose response of the viscous relaxation of radiation-sensitive gels to discuss their potential for dose imaging. Three batches of a radiation-sensitive polymer gel (MAGIC gel) samples were homogeneously irradiated to doses from 0 Gy to 45.5 Gy. Young's modulus was computed from the measured stress on the sample surface and the strain applied to the sample when compressing it axially, and the viscous relaxation was determined from the stress decay under sustained compression. The viscous relaxation was found not to change significantly with dose. However, Young's modulus was dose dependent; it approximately doubled in the gels between 0 Gy and 20 Gy. By fitting a second-order polynomial to the Young's modulus-versus-dose data, 99.4% of the variance in Young's modulus was shown to be associated with the change in dose. The precision of the gel production, irradiation and Young's modulus measurement combined was found to be 4% at 2 Gy and 3% at 20 Gy. Potential sources of measurement error, such as those associated with the boundary conditions in the compression measurement, inhomogeneous polymerization, temperature (up to 1% error) and the evaporation of water from the sample (up to 1% error), were estimated and discussed. It was concluded that Young's modulus could be used for dose determination. Imaging techniques such as elastography may help to achieve this if they can provide a local measurement of Young's modulus, which may eliminate problems associated with the boundaries (e.g. variation in coefficient of friction) and inhomogeneous polymerization. Elastography combined with a calibration should also be capable of mapping dose in three dimensions.

A kind of novel biodegradable supramolecular hydrogel was synthesized via copolymerization of gelatin methacrylamide with photocurable and biodegradable polypseudorotaxanes under UV irradiation. These polypseudorotaxanes were prepared by supramolecular self-assemblies of alpha-cyclodextrins threaded onto amphiphilic LA-PEG-LA copolymers end-capped with methacryloyl groups. The hydrogels are injectable, and their structure was characterized in detail with FTIR, (1)H NMR, XRD, TG and DSC techniques. Their swelling behaviour and morphologies were also examined. The analytical results demonstrated that the channel-type crystalline structure of the polypseudorotaxanes remains in the as-obtained hydrogels. Moreover, the SEM pictures showed that the hydrogels having gelatin methacrylamide are more suitable for cell seeding and proliferation than those without gelatin added.

The effects of electron beam or gamma-irradiation on technological performances (capsule hardness, expressed as deforming work and dissolution time) of empty 2-shell capsules made of gelatin or hydroxypropylmethylcellulose (HPMC) were studied. Capsule structural changes induced by radiation treatment were investigated by capillary viscometry and atomic force microscopy (AFM). The capsules were irradiated in the air at 5, 15, and 25 kGy. The deforming work of nonirradiated HPMC capsules (0.06 +/- 0.01 J) was lower than that of gelatin capsules (0.10 +/- 0.01 J). The dissolution time of the HPMC capsules (414 +/- 33 seconds) was slightly higher than that determined for gelatin hard capsules (288 +/- 19 seconds). The hardness and dissolution time of gelatin and HPMC capsules were not significantly influenced by the irradiation type and the applied irradiation dose. As the viscometry analyses are concerned, irradiation caused a reduction of the intrinsic viscosity and water and dimethyl sulfoxide solvent power in both the cases. AFM analysis showed that the radiation treatment did not appreciably affect the surface roughness of the samples nor induce structural changes on capsule surface. However, measurements of force-distance curves pointed out a qualitative parameter for the identification of the irradiated capsules. On the bases of these preliminary results, empty gelatin or HPMC hard capsules can be sanitized/sterilized by ionizing radiation.

With the introduction of conformal techniques in radiation therapy, gel dosimetry plays an important role as a 3D dose verification system. There are two main types of gels in use for dosimetry: Fricke gels and polymer gels. The advantages of polymer gels are improved dose response and stability with no diffusion problems. However, the more complicated fabrication procedure and the greater cost compared to Fricke gels makes polymer gels less attractive in routine clinical use. Dose resolution has recently been introduced as a concept for comparing and optimizing the performance of different types of gel dosimeters. This parameter has not yet been investigated for Fricke gels. In this study, the effect on the dose resolution and the diffusion from different gelatine- and Fe2+-concentrations and different pH was evaluated. Increasing the concentration of gelatine from 6 wt% to 10 wt% influenced the diffusion coefficient the most, while reducing the pH from 2.0 to 1.5 had the largest effect on the dose resolution. For a gel consisting of 10 wt% gelatine, 1.0 mM Fe2+ and pH 1.5 the diffusion coefficient was found to be 1.5 mm2 h-1 and the dose resolution was about 4.1% (at 95% confidence level), for a dose of 40 Gy. By evaluating different dose gradients by the gamma-method, the diffusion was shown to have no clinically relevant impact on the dose distribution and plan acceptance within 3 h of irradiation. The results indicate a potential use of Fricke gels for IMRT verification.

A high-frequency (85 MHz) acoustic technique is used to identify system parameters for controlled laser-induced microbubble creation inside tissue-mimicking, gelatin phantoms. Microbubbles are generated at the focus of an ultrafast 793-nm laser source and simultaneously monitored through ultrasonic pulse-echo recordings. Displayed in wavefield form, these recordings illustrate microbubble creation, and integrated backscatter plots provide specifics about microbubble characteristics and dissolution behavior. By varying laser parameters, including pulse fluence (or pulse energy flux, J/cm2), total number of pulses delivered, and the period between pulses, the size, lifetime, and dissolution dynamics of laser-induced microbubbles may be independently controlled. Pulse fluence is the main size-controlling parameter, whereas both increases in pulse fluence and pulse number can lengthen microbubble lifetime from tens to hundreds of milliseconds. In short, a microbubble of particular lifetime does not necessarily have to be of a particular size. Microbubble behavior, furthermore, is independent of pulse periods below a fluence-dependent threshold value, but it exhibits stochastic behavior if pulse repetition is too slow. These results demonstrate that laser pulse fluence, number, and period may be varied to deposit energy in a specific temporal manner, creating and stabilizing microbubbles with particular characteristics and, therefore, potential uses in sensitive acoustic detection and manipulation schemes.

Gelatin having p-nitrocinnamate pendant groups (Gel-NC) was prepared via an efficient one-pot synthesis, yield >87%. (1)H NMR data indicated that 1 mol of gelatin was modified with 18 +/- 6 mol of the photosensitive group. Upon exposure to low-intensity 365 nm UV light and in the absence of photoinitiators or catalysts, Gel-NC cross-linked within minutes into a gelatin-based hydrogel as monitored by UV-vis spectroscopy. The degree of swelling of this biodegradable hydrogel in aqueous solutions responded to changes in Gel-NC concentration levels, the ionic strength of the aqueous solutions, and photo-cross-linking time. Topography changes associated with phase transition resulting from "photocleavage" of the hydrogel network with 254 nm UV light were studied with AFM. Both Gel-NC and its hydrogel expressed low toxicity to human neonatal fibroblast cells. In addition, gelatin-based microgels were prepared via the photo-cross-linking of Gel-NC within inverse micelles.

Photoinduced transformation in gelatin films made with 14-fluoro bacteriorhodopsin derivatives, both wild-type (WT) and D96N mutant, were studied. Spectral and kinetic peculiarities for these two types of samples were compared over a wide range of relative humidity (9-92%). Analysis of the results considered two existing photoinduced processes that occur in suspensions and films of corresponding pigments. It was demonstrated that there is a range of humidity in which the performance of fluorine WT bacteriorhodopsin gelatin films may offer a technological advantage compared with fluorine D96N bacteriorhodopsin.

This study is an investigation to evaluate the in vivo degradation of gelatin hydrogels in terms of their number of cross-links. Various hydrogels were prepared from acidic gelatin, extracted from bovine bone, porcine skin or fish scale, and basic gelatin, extracted from porcine skin, through four types of cross-linking methods, i.e., glutaraldehyde (GA) or dehydrothermal treatment and ultraviolet (UV) or electron beam irradiation. The water content of hydrogels and their number of cross-links, calculated from the tensile modulus of hydrogels, were evaluated as the measure of hydrogel cross-linking extent. Following subcutaneous implantation of 125I-labeled gelatin hydrogels into mice, the radioactivity remaining was measured at different time intervals to assess the in vivo degradability of hydrogels. Irrespective of the gelatin type and cross-linking method, a good correlation was found between the in vivo degradability of hydrogels and their number of cross-links, which is different from the correlation to their water content. This finding indicates that the degradability of hydrogels is governed by their number of cross-links.

We devised a new transtissue drug-delivery system, based on a multiple-needle-arrayed injector that has 36 long and short needles on the needle head, to administer the drug into local points of the target tissue at a well-controlled depth and pitch. A preliminary in vitro study, focusing the time-dependent depth profiling of protein injected in agarose gel as a model tissue using confocal laser scanning microscope, was conducted to evaluate the performance of the multiple-needle-arrayed injector coupled with photoreactive gelatin (styrenated gelatin: St-gelatin) as the sustained-release vehicle. Rhodamine-conjugated albumin, which was mixed with the St-gelatin buffer solution, was the model drug of the in vitro study, and the mixture was injected into agarose gel using the multiple-needle-arrayed injector by single injection, followed by visible-light irradiation to photocure the gelatin solution. Time-dependent distribution from the injected material into the surrounding agarose gel was observed using a confocal laser scanning microscope up to seven days. Injection of the drug material and concomitant withdrawal of the syringe (termed multirod method) enabled the long- and short-rod-like injections into the agarose gel at the same locations of the injected sites. The model drug gradually diffused throughout the agarose gel. In an in vivo study, the comparison of the efficacy of the angiogenic protein (bFGF: 10 microg for each) with placebo was performed using the non-ischemic hind limb model of rabbits. Four weeks after injection, a significant increase in the number of angiogenic capillaries was observed in the mixed St-gelatin/bFGF group compared with that of placebo. The multiple-needle-arrayed injector coupled with a sustained-release vehicle may be an effective drug delivery system for realizing the spatio-regional distribution of angiogenic protein.

To investigate the effect of immobilized cytokine, erythropoietin (Epo) was immobilized on a culture plate and the Epo-dependent human leukemia cell line UT-7/Epo then was cultured upon the plate. A photo-reactive gelatin was mixed with Epo and the mixture was cast on a plate. The plate was then irradiated with ultraviolet light in the presence or absence of a photo-mask. After washing with water, a micropatterned or unpatterned surface was formed. A leukemia cell line dependent on Epo, UT-7/Epo, was cultured on the sample plate. On the micropatterned surface, apoptosis of cells was induced on the surface without Epo, but was not observed on the Epo-immobilized surface. This result demonstrated that Epo stimulated the cells even after immobilization. Although the activity of immobilized Epo was low, the activity was slightly higher than that achieved by soluble Epo at higher concentration. In addition, the immobilized Epo could be repeatedly used for culture of UT-7/Epo cell. The present study provided a convenient immobilization method and indicated that immobilization of cytokines will be useful for creating an artificial cell culture device.

Selective hyperthermia can be a feasible treatment modality for deep tissue abnormalities. It is accomplished by using a laser or ultrasound noninvasively to transfer energy to a desired target causing tissue damage. This process has two potential benefits to medical professionals: simplicity of procedure and safety to patient. However, optimizing these selective interactions is difficult due to the number of variables. We propose an optimization coefficient relating the dynamic and geometric parameters of selective hyperthermia, and proceed to measure it in an experimental setup consisting of a near-infrared laser and laser-absorbing dye. To simulate tissue, gelatin phantoms are created using a combination of water, intralipid, and gelatin. Our experiments use a 1.00-cm-diam spherical phantom that is homogeneously enhanced with an indocyanine green (ICG) solution and placed inside a nontarget phantom and irradiated by an 805-nm diode laser. Temperature measurements taken at different locations are analyzed so optimization coefficients can be calculated for different parameters. This optimization coefficient compares the difference in temperatures from inside and outside the target. Analysis of the values after thermal equilibrium provides information about the best parameter selection. Our findings indicate that the optimal ICG concentration and power combination for our tested parameters are 0.083% and 0.97 W, respectively. Based on our analysis, optimization can be obtained by using this coefficient to compare the selectivity of several parameter combinations., ((c) 2004 Society of Photo-Optical Instrumentation Engineers.)

The crosslinking process of natural macromolecules with microwave energy should have the potentiality to overcome the problems due to the toxicity of the residuals of chemical crosslinking agents and moreover of the "in vivo" biodegradation products of the chemical crosslinked macromolecule. To evaluate the effective crosslinking of the gelatin forming the microspheres, the water-soluble fraction at 37 degrees C, the water absorption capability, the free amino and free carboxylic acid groups of the gelatin were determined. The structural change in the gelatin microspheres has been detected by the porosity studies. Moreover, both the "in vitro" biodegradability and the biocompatibility of the gelatin microspheres microwave-treated after a subcutaneous injection into female albino guinea pigs were tested. As the results suggest only the gelatin microspheres microwave-treated for 10 min at an inlet temperature of 250 degrees C could have been modified by the crosslink formation among the macromolecular chains. The gelatin microspheres treated with the microwave energy were very well biodegraded as indicated both by the "in vitro" enzymatic degradation studies and mainly by the histopathological examination. This latter study has also demonstrated the biocompatibility of the gelatin microspheres crosslinked with the microwave energy. In order to evaluate the feasibility of the microwave crosslinking process for pharmaceutical applications, both the drug loading and the drug release processes were evaluated using diclofenac as drug model, either as acidic form or as sodium salt. The microspheres were swollen in aqueous solution of diclofenac sodium salt, followed by a washing procedure with cool water to maintain the sodium salt into the microspheres or with pH 1.5 HCl to induce the diclofenac precipitation. To increase the amount of diclofenac acid form in the microspheres, the procedure was repeated three times washing with pH 1.5 HCl after each swelling process. Both the X-ray diffractometry and thermal analysis investigations showed a different physical state of the two drug forms in the microspheres, i.e. the amorphous state of the sodium salt and the crystalline state of the acidic form. According to the experimental results, the drug is released from gelatin microspheres according to the drug loading and the drug solubility.

Purpose: Although gemcitabine, a deoxycytidine analogue, recently demonstrated improvements in the response rate for pancreatic cancer, the median survival for patients is limited to 4-6 months. The purpose of the present study was to develop trans-tissue delivery of gemcitabine, which is based on photocured gelatin gel immobilized with gemcitabine, and to validate whether such a system inhibits the growth of the pancreatic tumor in vivo., Experimental Design: The in vitro release profile of gel-embedded gemcitabine from a gel was examined based on in vitro chemosensitivity of AsPC1 cell (human pancreatic cancer cell line) for gemcitabine. The permeation of gel-embedded rhodamine B (used as a model drug) into tissues and inhibitory effect of tumor growth of photocured gelatin gel immobilized with gemcitabine were examined using in vivo s.c. tumor model of athymic mice., Results: The release profile was characterized as an initial burst of release, followed by a gradual release, irrespective of gelatin concentration. Rhodamine B permeated into the tumor and retained for at least 10 days. Photocured gelatin gel immobilized with gemcitabine significantly reduced the tumor volume compared with gemcitabine injection. Therapeutic success was correlated with decreased cell proliferation and increased cell apoptosis in tumor cells, supported by proliferating cell nuclear antigen and terminal deoxynucleotidyltransferase-mediated nick end labeling staining. Blood analysis and body weight measurement showed that little side effect was observed in this therapy., Conclusions: In situ trans-tissue gemcitabine delivery on the tissue with possibly remnant cancer cells using the drug-releasing matrix developed here is expected to reduce the rate of local recurrence for patients with pancreatic cancer.

Magnetization transfer (MT) has been measured quantitatively as a function of radiation dose in MAGIC polymer gels. The MT rates between the free and immobile macromolecular proton pools (kmr and kfm), and the ratio of the sizes of these coupled proton pools (Pm/Pf), were measured by analysing the response to an inversion recovery sequence. While pm/pf increases linearly with dose, the fast MT rate kmf also increases with dose, unlike previous measurements in BANG gels. This dependence of kmf on dose suggests there are additional factors that modify spin exchange in MAGIC gels as irradiation occurs.

A new type of polymer gel dosimeter, which responds well to absorbed dose even when manufactured in the presence of normal levels of oxygen, was recently described by Fong et al. [Phys. Med. Biol. 46, 3105-3113 (2001)] and referred to by the acronym MAGIC. The aim of this study was to investigate the feasibility of using this new type of gel for intensity-modulated radiation therapy (IMRT) verification. Gel manufacturing was carried out in room atmosphere under normal levels of oxygen. IMRT inverse treatment planning was performed using the Helios software. The gel was irradiated using a linear accelerator equipped with a dynamic multileaf collimator, and intensity modulation was achieved using sliding window technique. The response to absorbed dose was evaluated using magnetic resonance imaging. Measured and calculated dose distributions were compared with regard to in-plane isodoses and dose volume histograms. In addition, the spatial and dosimetric accuracy was evaluated using the gamma formalism. Good agreement between calculated and measured data was obtained. In the isocenter plane, the 70% and 90% isodoses acquired using the different methods are mostly within 2 mm, with up to 3 mm disagreement at isolated points. For the planning target volume (PTV), the calculated mean relative dose was 96.8 +/- 2.5% (1 SD) and the measured relative mean dose was 98.6 +/- 2.2%. Corresponding data for an organ at risk was 34.4 +/- 0.9% and 32.7 +/- 0.7%, respectively. The gamma criterion (3 mm spatial/3% dose deviation) was fulfilled for 94% of the pixels in the target region. Discrepancies were found in hot spots the upper and lower parts of the PTV, where the measured dose was up to 11% higher than calculated. This was attributed to sub optimal scatter kernels used in the treatment planning system dose calculations. Our results indicate great potential for IMRT verification using MAGIC-type polymer gel.

In order to obtain a gelatin hydrogel crosslinked by a reagent-free method, gamma-ray and electron beam radiation was applied to porcine, bovine and fish gelatin gels and the products were characterized by measuring the gel fraction, the swelling ratio and the enzymatic degradability. On increasing the radiation dose, the gel fraction increased and both the swelling ratio and the enzymatic degradability decreased. The transition temperature from gel to sol of the hydrogel containing more than 5% mammal gelatins increased up to more than 90 degrees C when gamma-ray or electron beam were irradiated by more than 10 kGy. The results show that the degree of crosslinking of irradiated gelatin hydrogels increases with increasing irradiation dose and with decreasing concentration. It is suggested that the radiation crosslinking occurs around the physical crosslinking point or multiple helix structure of gelatin gel.

Ultrasonic speed of propagation and attenuation were investigated as a function of absorbed radiation dose in PAG and MAGIC polymer gel dosimeters. Both PAG and MAGIC gel dosimeters displayed a dependence of ultrasonic parameters on absorbed dose with attenuation displaying significant changes in the dose range investigated. The ultrasonic attenuation dose sensitivity at 4 MHz in MAGIC gels was determined to be 4.7 +/- 0.3 dB m(-1) Gy(-1) and for PAG 3.9 +/- 0.3 dB m(-1) Gy(-1). Ultrasonic speed dose sensitivities were 0.178 +/- 0.006 m s(-1) Gy(-1) for MAGIC gel and -0.44 +/- 0.02 m s(-1) Gy(-1) for PAG. Density and compressional elastic modulus were investigated to explain the different sensitivities of ultrasonic speed to radiation for PAG and MAGIC gels. The different sensitivities were found to be due to differences in the compressional elastic modulus as a function of dose for the two formulations. To understand the physical phenomena underlying the increase in ultrasonic attenuation with dose, the viscoelastic properties of the gels were studied. Results suggest that at ultrasonic frequencies, attenuation in polymer gel dosimeters is primarily due to volume viscosity. It is concluded that ultrasonic attenuation significantly increases with absorbed dose. Also, the ultrasonic speed in polymer gel dosimeters is affected by changes in dosimeter elastic modulus that are likely to be a result of polymerization. It is suggested that ultrasound is a sufficiently sensitive technique for polymer gel dosimetry.

Neural retinal transplantation has great potential for the alleviation of different degenerative and hereditary retinal disorders. However, because of the fragile and soft nature of retina, retinal-sheet transplantation is relatively difficult to achieve. To overcome this difficulty, we developed a technique for lamellar tissue transplantation. Biodegradable gelatin membranes were fabricated into a sandwich and encapsulated retinal grafts for transplantation. Before transplantation, we characterized the in vivo and in vitro properties of such membranes to determine the optimal sterilization procedure, that is, a sterile membrane with suitable degradability and good mechanical properties and without cytotoxicity. Three sterilization methods were conducted, with hydrogen peroxide gas plasma (HPGP), ethylene oxide (EO), and gamma-ray irradiation (gamma). The results were compared with those of a control (no disinfection). Initial studies revealed that the gelatin membranes sterilized with HPGP or EO exhibited retinal pigment epithelium (RPE) cytotoxicity, whereas the membrane sterilized by 16.6-kGy gamma ray irradiation had no RPE cytotoxicity and had enhanced mechanical properties. In the in vivo rabbit study, implanted gelatin membranes demonstrated satisfactory biocompatibility without any inflammation. Transplanted retinal sheets survived well and developed laminar structures. Such a method using gelatin membranes for tissue transportation has great potential for future routine retinal-sheet transplantation., (Copyright 2002 Wiley Periodicals, Inc.)

Background/objective: Laser thrombolysis is the selective removal of thrombus from occluded blood vessels using laser energy. A reconstituted clot model with reproducible optical absorption properties was developed to evaluate the effect of various laser parameters on thrombus removal rate., Study Design/materials and Methods: Reconstituted clots were made with known fibrinogen concentrations and hematocrits. Ex vivo clots were collected from ten swine. Four red gelatin phantoms were prepared. Mass removal rates and ablation efficiencies were determined using a 577 nm, 1 microsec pulsed dye laser. The ablation efficiencies of the three clot models were compared at an energy of 25 mJ and a repetition rate of 4 Hz. In addition, the reconstituted clot model was ablated as pulse energy and repetition rate were varied with average power held constant at 100 mW., Results: The mean ablation efficiency for ex vivo clots ranged from 0.4 +/- 0.1 to 3.4 +/- 0.7 microg/mJ/pulse, with significant differences between groups (ANOVA p < 0.05). Reconstituted clots of varied fibrinogen content had ablation efficiencies of 1.5 +/- 0.2 to 1.6 +/- 0.3 microg/mJ/pulse at this energy and repetition rate. Gelatin ablation efficiency was inversely proportional to protein content and ranged from 0.5 +/- 0.3 to 2.0 +/- 0.7 microg/mJ/pulse. Reconstituted clot mass removal rates (in microg/s) were clinically similar for settings ranging from 13 mJ at 8 Hz to 33 mJ at 3 Hz., Conclusions: The reconstituted model clot is a reproducible and biologically relevant thrombolysis target. Ex vivo clot lacks reproducibility between individuals and gelatin phantoms lack clinical relevance. At a constant average power, varying laser parameters did not affect mass removal rates to a clinically significant degree.