Modification Of Gelatin Research Articles

Microfluidic-enabled bottom-up hydrogels from annealable naturally-derived protein microbeads

Naturally-derived proteins, such as collagen, elastin, fibroin, and gelatin (denatured collagen) hold a remarkable promise for tissue engineering and regenerative medicine. Gelatin methacryloyl (GelMA), synthesized from the methacryloyl modification of gelatin, mimicking the structure of extracellular matrix, has widely been used as a universal multi-responsive scaffold for a broad spectrum of applications, spanning from cell therapy to bioprinting and organoid development. Despite the widespread applications of GelMA, coupled stiffness and porosity has inhibited its applications in 3D cellular engineering wherein a stiff scaffold with large pores is demanded (e.g., at concentrations >10 wt%). Taking advantage of the orthogonal thermo-chemical responsivity of GelMA, we have developed microfluidic-assisted annealable GelMA beads, that are first stabilized by temperature-mediated physical crosslinking, flowed to form a scaffold structure, and then chemically annealed using light to fabricate novel bead-based 3D GelMA scaffolds with high mechanical resilience. We show how beaded GelMA (B-GelMA) provides a self-standing microporous environment with an orthogonal void fraction and stiffness, promoting cell adhesion, proliferation, and rapid 3D seeding at a high polymer concentration (∼20 wt%) that would otherwise be impossible for bulk GelMA. B-GelMA, decorated with methacryloyl and arginylglycylaspartic acid (RGD) peptide motifs, does not require additional functionalization for annealing and cell adhesion, providing a versatile biorthogonal platform with orthogonal stiffness and porosity for a myriad of biomedical applications. This technology provides a universal method to convert polymeric materials with orthogonal physico-chemical responsivity to modular platforms, opening a new horizon for converting bulk hydrogels to beaded hydrogels (B-hydrogels) with decoupled porosity and stiffness.

Structural Modification of Fish Gelatin by the Addition of Gellan, κ-Carrageenan, and Salts Mimics the Critical Physicochemical Properties of Pork Gelatin.

Certain people cannot consume any pork product or derivatives for religious reasons, thus it is essential to find a pork gelatin (PG) substitute for food product development. The commonly used polysaccharides, gellan and carrageenan, together with salt, can be added to fish gelatin (FG) to match the textural properties of PG, representing a promising substitute for PG. The difference in the mechanism of gellan and carrageenan to improve properties of FG has been revealed from nanostructure level. The use of food grade ingredients and simple mixing process are favorable in the food industry.

Epigallocatechin Gallate-Modified Gelatin Sponges Treated by Vacuum Heating as a Novel Scaffold for Bone Tissue Engineering.

Chemical modification of gelatin using epigallocatechin gallate (EGCG) promotes bone formation in vivo. However, further improvements are required to increase the mechanical strength and bone-forming ability of fabricated EGCG-modified gelatin sponges (EGCG-GS) for practical applications in regenerative therapy. In the present study, we investigated whether vacuum heating-induced dehydrothermal cross-linking of EGCG-GS enhances bone formation in critical-sized rat calvarial defects. The bone-forming ability of vacuum-heated EGCG-GS (vhEGCG-GS) and other sponges was evaluated by micro-computed tomography and histological staining. The degradation of sponges was assessed using protein assays, and cell morphology and proliferation were verified by scanning electron microscopy and immunostaining using osteoblastic UMR106 cells in vitro. Four weeks after the implantation of sponges, greater bone formation was detected for vhEGCG-GS than for EGCG-GS or vacuum-heated gelatin sponges (dehydrothermal cross-linked sponges without EGCG). In vitro experiments revealed that the relatively low degradability of vhEGCG-GS supports cell attachment, proliferation, and cell–cell communication on the matrix. These findings suggest that vacuum heating enhanced the bone forming ability of EGCG-GS, possibly via the dehydrothermal cross-linking of EGCG-GS, which provides a scaffold for cells, and by maintaining the pharmacological effect of EGCG.

Antimicrobial activity of organic–inorganic hybrid films based on gelatin and organomodified silicones

AbstractGelatin–siloxane hybrid films containing antimicrobial agents were prepared in the two‐step procedure. First, gelatin was chemically modified with organomodified silicones containing epoxy (S1) and both epoxy and fluoroalkyl groups (S2). Next, antimicrobial agents such as lysozyme, EDTA, and nisin, in various concentrations, were introduced into obtained hybrid solution, prior to solvent casting of the films. Antimicrobial activity of prepared samples, as well as neat gelatin and gelatin–siloxane hybrid as a reference, was investigated using a modified agar diffusion method against several bacterial strains (Aeromonas hydrophila, Bacillus subtilis, Escherichia coli) and opportunistic fungus (yeast species Candida albicans). Prepared gelatin–siloxane films were characterized using FT‐IR, thermal and SEM analysis. Mechanical properties and swelling behavior of the samples were also investigated. The influence of type and amount of organosilicon compound used for modification of gelatin, as well as type and concentration of applied active agent on antimicrobial activity, was determined. We observed higher antimicrobial activity for all synthesized gelatin–siloxane hybrids than for unmodified gelatin with lysozyme, nisin, and EDTA prepared as comparison. Moreover, samples obtained with siloxane containing fluoroalkyl groups (S2) showed higher resistance to microbial agents than hybrids with S1 siloxane. Hybrid films showed also better water stability than neat biopolymer.

Poly (acrylic acid) grafted gelatin nanocarriers as swelling controlled drug delivery system for optimized release of paclitaxel from modified gelatin

The present study was undertaken to carry out modification of gelatin by grafting of poly (acrylic acid) onto the backbone of gelatin via free radical polymerization of acrylic acid using potassium persulphate (KPS) as an initiator. The modified gelatin was transformed into nanoparticles following microemulsion crosslinking method and the prepared nanoparticles were investigated as swelling controlled drug delivery system for controlled release of paclitaxel. The structure, morphology and composition of the grafted gelatin nanoparticles (gelatin-g-PAA) were studied using various analytical techniques like FTIR, TEM, SEM, XRD, and zeta potential measurements, and in vitro cytotoxicity of nanoparticles was assayed. The influence of experimental conditions such as chemical composition of poly (acrylic acid) grafted gelatin nanoparticles, pH of the release media, presence of salt and sugar solutions were also investigated on the release profiles of paclitaxel. The data of the drug release processes were analysed kinetically using Ficks power law, zero order, first order and Korsmeyer- Peppas models and a correlation was drawn between the quantity of released drugs and degree of swelling of the nanoparticles.

Composite Colloidal Gels Made of Bisphosphonate‐Functionalized Gelatin and Bioactive Glass Particles for Regeneration of Osteoporotic Bone Defects

AbstractInjectable composite colloidal gels are developed for regeneration of osteoporotic bone defects through a bottom‐up assembly from bisphosphonate‐functionalized gelatin and bioactive glass particles. Upon bisphosphonate functionalization, gelatin nanoparticles show superior adhesion toward bioactive glass particles, resulting in elastic composite gels. By tuning their composition, these composite colloidal gels combine mechanical robustness with self‐healing ability. The composite colloidal gels support cell proliferation and differentiation in vitro without requiring any osteogenic supplement. In vivo evaluation of the composite colloidal gels reveals their capacity to support the regeneration of osteoporotic bone defects. Furthermore, the bisphosphonate modification of gelatin induces a therapeutic effect on the peri‐implantation region by enhancing the bone density of the osteoporotic bone tissue. Consequently, these composite colloidal gels offer new therapeutic opportunities for treatment of osteoporotic bone defects.

Fabrication of Highly Crosslinked Gelatin Hydrogel and Its Influence on Chondrocyte Proliferation and Phenotype.

Gelatin methacrylate (GelMA) hydrogels have been widely studied for biomedical applications, such as tissue engineering and drug delivery, because of their good biocompatibility and injectability. However, the quick degradation and low mechanical property of GelMA hydrogels need to be improved for further applications, especially for long-term implantation. In this study, a sequential double modification of gelatin was used to achieve high density of photocrosslinkable double bonds in gelatin derivatives. The amino groups in gelatin were first reacted with methacrylic anhydride. After this, the hydroxyl and carboxyl groups in gelatin were reacted with glycidyl methacrylate to obtain the double modified gelatin macromer. The double modified gelatin macromer was used to prepare gelatin hydrogels with high crosslinking density. The hydrogels exhibited high storage modulus and low degradation. Culture of bovine articular chondrocytes in the gelatin hydrogels showed that chondrocytes had round morphology and maintained a cartilaginous phenotype while cell proliferation was hampered. This method for increasing crosslinking density should be useful for preparation of stable hydrogels for cartilage tissue engineering.

Structural analysis of photocrosslinkable methacryloyl-modified protein derivatives.

Biochemically modified proteins have attracted significant attention due to their widespread applications as biomaterials. For instance, chemically modified gelatin derivatives have been widely explored to develop hydrogels for tissue engineering and regenerative medicine applications. Among the reported methods, modification of gelatin with methacrylic anhydride (MA) stands out as a convenient and efficient strategy to introduce functional groups and form hydrogels via photopolymerization. Combining light-activation of modified gelatin with soft lithography has enabled the materialization of microfabricated hydrogels. So far, this gelatin derivative has been referred to in the literature as gelatin methacrylate, gelatin methacrylamide, or gelatin methacryloyl, with the same abbreviation of GelMA. Considering the complex composition of gelatin and the presence of different functional groups on the amino acid residues, both hydroxyl groups and amine groups can possibly react with methacrylic anhydride during functionalization of the protein. This can also apply to the modification of other proteins, such as recombinant human tropoelastin to form MA-modified tropoelastin (MeTro). Here, we employed analytical methods to quantitatively determine the amounts of methacrylate and methacrylamide groups in MA-modified gelatin and tropoelastin to better understand the reaction mechanism. By combining two chemical assays with instrumental techniques, such as proton nuclear magnetic resonance (1H NMR) and liquid chromatography tandem-mass spectrometry (LC-MS/MS), our results indicated that while amine groups had higher reactivity than hydroxyl groups and resulted in a majority of methacrylamide groups, modification of proteins by MA could lead to the formation of both methacrylamide and methacrylate groups. It is therefore suggested that the standard terms for GelMA and MeTro should be defined as gelatin methacryloyl and methacryloyl-substituted tropoelastin, respectively, to remain consistent with the widespread abbreviations used in literature.

Novel electrospun nanofibers of modified gelatin-tyrosine in cartilage tissue engineering

In natural cartilage tissues, chondrocytes are linked to extracellular matrix (ECM) through cell-surface binding proteins. Surface modification of gelatin can provide a new generation of biopolymers and fibrous scaffolds with chemical, mechanical, and biological properties. In this study tyrosine protein and 1,2,3-triazole ring were utilized to functionalize gelatin without Cu catalyst. Their molecular structure was characterized by Fourier transform infrared spectroscopy (FTIR) and nuclear magnetic resonance spectroscopy (1HNMR). Chemical cross-linkers such as glutaraldehyde (GA) and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC)/N-hydroxysulfosuccinimide (NHS) were used to electrospin the modified gelatin. The modification of gelatin and cross-linking effects were confirmed by scanning electron microscopy (SEM), contact angle measurement, and mechanical tests. MTT assay using chondrocyte cells showed cell viability of electrospun modified gelatin scaffolds. In vitro cell culture studies showed that electrospun engineered protein scaffolds would support the attachment and growth of cells. The results also showed that cross-linked nanofibers with EDC/NHS could be considered excellent matrices in cell adhesion and proliferation before electrospinning process and their potential substrate in tissue engineering applications, especially in the field of cartilage engineering.

Biopolymer-based functional inks for the preparation of artificial cartilage via bioprinting technology

We present bioinks based on biopolymers from the native extracellular matrix, namely gelatin and chondroitin sulfate, which are suitable for bioprinting technology applying living mammalian cells. The bioinks can be crosslinked due to chemical modification of the biopolymers with methacrylic functions. Such insoluble hydrogels with tunable physico-chemical propertie are able to closely mimic the native extracellular matrix of biological tissues (ECM). Highly methacrylated gelatin (GM10) resulted in solutions with low viscosities (e.g. 15 wt%: 6.7 ± 1.0 mPa s at 37 °C) within the inkjet-printable range and crosslinked hydrogels with high mechanical strength. Less methacrylated gelatin (GM2) was proper for preparation of soft hydrogels and highly viscous bioinks for pneumatic dispensing technology (e.g. 15 wt%: 39.3 ± 3.6 mPa s at 37 °C). By additional acetylation of GM2 (GM2A8) the solution viscosity could be significantly lowered (e.g. 15 wt%: η = 5.2 ± 0.5 mPa s, 37 °C) and, thus, inkjet-printable bioinks with low crosslinking capacity were also gained. Consequently, a versatile bioink system for bioprinting technology was created. by adjustable twofold modification of gelatin with photochemically reactive and inert groups. Introduction of an additional methacrylated component of the ECM, methacrylated chondroitin sulfate (CSM), further tuned the chemical and physical properties of bioink and hydrogels. GM(A)-CSM hybrid hydrogels possessed for example significantly higher swellability than pure GM(A) hydrogels of the same mass fraction. CSM also influenced bioink viscosity: For unmodified gelatin and gelatin with low degree of modification CSM induced a decrease in bioink viscosity (e.g. 15 wt% GM2: 39.3 ± 3.6 mPa s versus 14 wt% GM2 + 1 wt% CSM: 32.9 ± 6.5 mPa s). In contrast to this, addition of CSM to bioinks made of gelatins with high degree of modification, GM2A8 or GM10, induced an increase in solution viscosity (e.g. 15 wt% GM2A8: 5.2 ± 0.5 mPa s versus 14 wt% GM2A8 + 1 wt% CSM: 6.5 ± 1.0 mPa s). The developed bioinks were proven to be suitable for bioprinting with viable mammalian cells, in this case porcine articular chondrocytes, and for fabrication of tissue models with intrinsic structure. Thus, biomaterials presented in this study can be used for biofabrication of artificial three-dimensional tissues with biomimetic organization, such as articular cartilage with its hierarchical structure.

Gelatin methacrylate microspheres for controlled growth factor release

Gelatin has been commonly used as a delivery vehicle for various biomolecules for tissue engineering and regenerative medicine applications due to its simple fabrication methods, inherent electrostatic binding properties, and proteolytic degradability. Compared to traditional chemical cross-linking methods, such as the use of glutaraldehyde (GA), methacrylate modification of gelatin offers an alternative method to better control the extent of hydrogel cross-linking. Here we examined the physical properties and growth factor delivery of gelatin methacrylate (GMA) microparticles (MPs) formulated with a wide range of different cross-linking densities (15–90%). Less methacrylated MPs had decreased elastic moduli and larger mesh sizes compared to GA MPs, with increasing methacrylation correlating to greater moduli and smaller mesh sizes. As expected, an inverse correlation between microparticle cross-linking density and degradation was observed, with the lowest cross-linked GMA MPs degrading at the fastest rate, comparable to GA MPs. Interestingly, GMA MPs at lower cross-linking densities could be loaded with up to a 10-fold higher relative amount of growth factor than conventional GA cross-linked MPs, despite the GA MPs having an order of magnitude greater gelatin content. Moreover, a reduced GMA cross-linking density resulted in more complete release of bone morphogenic protein 4 and basic fibroblast growth factor and accelerated release rate with collagenase treatment. These studies demonstrate that GMA MPs provide a more flexible platform for growth factor delivery by enhancing the relative binding capacity and permitting proteolytic degradation tunability, thereby offering a more potent controlled release system for growth factor delivery.

Biofunctionalization of gelatin microcarrier with oxidized hyaluronic acid for corneal keratocyte cultivation

Development of microcarriers as alternative architecture models to traditional three-dimensional porous scaffolds can potentially eliminate the concern of extensive geographic necrosis during keratocyte cultivation. For the first time, this work was performed to evaluate the in vitro expansion of rabbit corneal keratocytes (RCKs) on the gelatin microcarrier (GMC) modified with hyaluronic acid (HA). In order to confirm the successful synthesis, the oxidized HA (oHA) samples following treatment of HA with sodium periodate were investigated by chemical characterization, including Fourier transform infrared and nuclear magnetic resonance spectroscopy. Biofunctionalization of GMC with oHA was then achieved by the formation of aldimine linkage between free amino groups in protein and aldehyde groups in oxidized polysaccharide. The grafting yield of oHA onto GMC determined by alcian blue staining assay was 26.9±0.1%. Tissue culture polystyrene (TCPS) plates and the microcarriers from either the GMC or GMC-oHA group were used for suspension culture of RCKs. Results of WST-1 assays, scanning electron microscopy, extracellular matrix production assays, and quantitative real-time reverse transcription polymerase chain reaction and Western blot analysis showed that among all the groups studied, the GMC-oHA samples could most effectively support a large scale growth of RCKs while enhancing their total collagen and glycosaminoglycan contents (i.e., indicative of biosynthetic capacity) and keratocan and ALDH1 gene and protein expressions (i.e., indicative of cell phenotype and function). It is concluded that the aforementioned keratocyte behaviors are found to be strongly dependent on the type of culture substrate (i.e., TCPS plate versus biopolymer microcarrier) and on the biomaterial functionalization (i.e., modification of gelatin with oHA).

Radiation stability and modification of gelatin for biological and medical applications

Gelatin is used in various biological and medical fields, including drug delivery systems and tissue engineering. In the context of these applications, radiation sterilization of gelatin was evaluated in terms of radiation stability. The molecular weight of gelatin powder irradiated by electron beams (EB) was analyzed using gel permeation chromatography (GPC). We found that irradiation decomposed the gelatin and that the weight-averaged molar mass (Mw) decreased by approximately 7–10% with sterilization doses in the range of 5–25kGy. Also, we found that the hydrolysis rate in body and cell culture environments (37°C water) was affected by irradiation. Although gelatin powder underwent chain scission when irradiated, crosslinking was predominantly induced when the gelatin was irradiated in water solution. Radiation-crosslinked (RX) gelatin hydrogel was fabricated without using any crosslinkers. In this case, fabrication and radiation sterilization were performed simultaneously. Using gel fraction and GPC analysis of the eluted sol, it was determined that the RX-gelatin hydrogel was stable for 7 days in water at 37°C. These results provide important data for evaluating the feasibility of biological and medical applications of gelatin and RX-gelatin hydrogel.

Gelatin-Based Materials in Ocular Tissue Engineering.

Gelatin has been used for many years in pharmaceutical formulation, cell culture and tissue engineering on account of its excellent biocompatibility, ease of processing and availability at low cost. Over the last decade gelatin has been extensively evaluated for numerous ocular applications serving as cell-sheet carriers, bio-adhesives and bio-artificial grafts. These different applications naturally have diverse physical, chemical and biological requirements and this has prompted research into the modification of gelatin and its derivatives. The crosslinking of gelatin alone or in combination with natural or synthetic biopolymers has produced a variety of scaffolds that could be suitable for ocular applications. This review focuses on methods to crosslink gelatin-based materials and how the resulting materials have been applied in ocular tissue engineering. Critical discussion of recent innovations in tissue engineering and regenerative medicine will highlight future opportunities for gelatin-based materials in ophthalmology.

Chemical tailoring of gelatin to adjust its chemical and physical properties for functional bioprinting.

Double chemical functionalization of gelatin by methacrylation and acetylation of free amino groups enables control over both the viscous behavior of its solutions and the mechanical properties of the resulting hydrogels after photochemical crosslinking. The degree of methacrylation is controlled by the molar excess of methacrylic anhydride applied. Tenfold molar excess leads to highly methacrylated gelatin (GM10), resulting in solutions with low viscosities within the inkjet-printable range (10 wt%: 3.3 ± 0.5 mPa s, 37 °C) and crosslinked hydrogels with high storage moduli G' (10 wt%: 15.2 ± 6.4 kPa). Twofold excess of methacrylic anhydride leads to less methacrylated gelatin (GM2) proper for preparation of soft hydrogels (10 wt%: G' = 9.8 ± 4.6 mPa s) but its solutions are highly viscous (10 wt%: 14.2 ± 1.1 mPa s, 37 °C) and thus prone to clogging printing nozzles. Here we show that additional introduction of acetyl functionalities into GM2 results in a significant decrease in solution viscosity (10 wt%: 2.9 ± 0.2 mPa s, 37 °C) and prevention of physical gel formation. In such a manner twofold functionalized gelatin can be inkjet-printed while the degree of chemical crosslinking remains low and the resulting gels are soft. Thus, by adjustable twofold modification of gelatin, i.e. inserting photochemically reactive and inert groups, a versatile bioink for inkjet bioprinting is created, which allows for addressing ECM based hydrogel matrices with a broad range of physical properties. Moreover, bioinks are proven to be cytocompatible and proper for inkjet printing of viable mammalian cells.

Stiff gelatin hydrogels can be photo-chemically synthesized from low viscous gelatin solutions using molecularly functionalized gelatin with a high degree of methacrylation

Gelatin is a very promising matrix material for in vitro cell culture and tissue engineering, e.g. due to its native RGD content. For the generation of medical soft tissue implants chemical modification of gelatin improves the mechanical properties of gelatin hydrogels and the viscous behavior of gelatin solutions for liquid handling. We present a systematic study on the influence of high degrees of methacrylation on the properties of gelatin solutions and photo-chemically crosslinked hydrogels. Changes from shear thinning to shear thickening behavior of gelatin solutions were observed depending on mass fraction and degree of methacrylation. Degrees of swelling of crosslinked hydrogels ranged from 194 to 770 % and storage moduli G' from 368 to 5 kPa, comparable to various natural tissues including several types of cartilage. Crosslinked gels proofed to be cytocompatible according to extract testings based on DIN ISO 10933-5 and in contact with porcine chondrocytes.

Glyoxal modification of gelatin leads to change in properties of solutions and resulting films

The influence of different pretreated film forming solutions (FFSs) on the physical, chemical and barrier properties of gelatin films was studied. Hydrolyzation with hydrochloric acid and acetylation with acetic acid anhydride at different pHs were performed. Subsequently, gelatin was crosslinked with glyoxal to result in biopolymer films with properties predetermined by the rheological behavior of the respective FFS. Gel time, storage modulus and tan δ of FFSs were obtained from small-strain oscillatory measurements and were correlated with swelling and solubility of the resulting films as well as with chemical analysis of lysine and arginine and the respective amino acid modifications Ne-carboxymethyllysine (CML), glyoxal derived lysine dimer (GOLD) and 5-(2-imino-5-oxo-1-imidazolidinyl)norvaline (imidazolinone). In general, chemical modification reduced the storage modulus whether it was hydrolyzation, acetylation or modification with glyoxal. Treatment by hydrolyzation or acetylation led to differences in gel time and in the ratio between viscous and elastic portion of the different hydrogels, also affecting the relationship between monovalent amino acid modification and bivalent crosslinking after glyoxal addition. Thus, the structural character of FFS hydrogels and of the protein network of films both determined the physical and barrier properties of chemically modified gelatin films.

Molecular and structural characteristics of cod gelatin films modified with EDC and TGase

Cod gelatin films before and after cross-linking of gelatin with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) or transglutaminase (TGase) have been characterized by Fourier transform infrared (FT-IR) spectroscopy and differential scanning calorimetry (DSC) analysis. For comparison, a film prepared from unmodified pig gelatin has been also analysed. The difference spectra showed that cod gelatin during the film formation involved first of all water-to-amide hydrogen bonds, and the film from pig gelatin contained water-to-amide, amide-to-amide and water-to-water hydrogen bonds. A higher number of hydrogen bonds in the structure of the film from pig gelatin contributed to much better recovering of the helical structure in this film than in the film from cod gelatin, as the peaks at about 1663 cm −1 in the amide I band and at about 1537 cm −1 in the amide II band in the second-derivative spectra revealed. The recovered helical structure, in turn, resulted in a significantly higher melting enthalpy value in the case of the film from the pig gelatin. After modification of cod gelatin with EDC or TGase, the inter-chain cross-linkages formed in the films led to the conformation of gelatin with no indications of helical ordering. An increase of melting temperature of gelatin films by 7 °C on EDC and by 10 °C on the TGase modifications was related to the formation of covalent cross-links, and a decrease of glass temperature by 28 °C and 7 °C on EDC and TGase cross-linking, respectively, demonstrated the plasticizing effect of water.

Emulsifying Property and Antioxidative Activity of Cuttlefish Skin Gelatin Modified with Oxidized Linoleic Acid and Oxidized Tannic Acid

Cuttlefish skin gelatins modified with oxidized linoleic acid (OLA) and oxidized tannic acid (OTA) were characterized and determined for emulsifying properties and antioxidative activity. Modification of gelatin with 5% OTA increased the total phenolic content and 1,1-diphenyl-2-picrylhydrazyl, 2,2′-azinobis(3-ethylbenzothiazoline-6-sulphonic acid) radical scavenging activity and ferric reducing antioxidant power of gelatin–OTA. Incorporation of OLA into gelatin (OLA-to-free amino group molar ratio of 10:1) increased surface hydrophobicity of gelatin from 17.39 to 32.38 and reduce surface tension at air/water interface of gelatin solution from 53 to 32 mN/m. Gelatin–OLA had the increase in emulsion activity index, compared with gelatin without modification and was capable of producing a fine emulsion (d32 = 0.79 μm, d43 = 0.82 μm). Modification of gelatin–OLA complex with OTA at different concentrations (2.5%, 5%, and 10%) increased antioxidative activity but decrease emulsifying properties. However, gelatin–OLA modified with 5% OTA had higher emulsifying properties than the commercial gelatin (bovine gelatin). The presence of an alkyl group and a hydroxyl group in gelatin after modification with OLA and OTA, respectively, was revealed by Fourier transform infrared study. Coincidental decrease in free amino group was also noticeable in modified gelatin. Menhaden oil-in-water emulsion stabilized by gelatin modified with OLA and 5% OTA was more resistant to lipid oxidation and phase separation as evidenced by the lower thiobarbituric acid reactive substances value and smaller oil droplet size, compared with that stabilized by commercial bovine gelatin. Thus, the modification of gelatin by both OLA and OTA was able to improve antioxidative and emulsifying properties of cuttlefish skin gelatin.

Chemically Modified Gelatin as Biomaterial in the Design of New Nanomedicines

The synthesis of new polymers has led to dramatic enhancements in the medical field. In particular, new chemical entities provided new prospects in tissue engineering, cellular therapy and drug delivery. However, significant efforts still need to be taken in consideration in order to achieve diverse clinical applications in these fields, which is challenging because of the lack of suitable materials with desired microstructure, permeability, degradation rates, products, and suitable mechanical properties. For these reasons some chemical strategies are focused in back to the nature approaches or, in other words, in improving the properties of natural polymers by chemical modifications. We report that by using a simple chemical modification technique we can obtain new biomaterials, specifically suitable for biomedical applications. Concretely, we describe the chemical modification of gelatin and the suitable characteristics of the modified protein to develop new nanomedicines. This protein was selected because of its enormous potential in biomedicine, which is currently limited due to the difficulty of its use without toxic chemical crosslinkers. The modification of the protein was based on the transformation of the carboxylic group into amido groups after their reaction with polyamines, leading to a positively charged biopolymer. To cationize the gelatin two polyamines where used: ethylenediamine and spermine, the latter being one of the endogenous polyamines which has a very positive influence over cells. This modification was monitored by physico-chemical techniques such as NMR, spectrophotometry and potentiometry. With the most promising modified gelatins we were able to develop nanoparticles using the ionotropic gelation technique. In order to determine the ability of these new nanosystems to associate bioactive molecules we selected a model plasmid DNA. The developed nanosystems were characterized corroborating their ability to associate the genetic material. In conclusion, we were able to obtain a semi-synthetic biomaterial with tunable physico-chemical properties, which can be used to develop new nanosystems with the ability to associate genetic material. We therefore propose that the gelatin, with its chemical modification, provide a unique biomaterial for the development of new nanomedicines.