Chitosan Gelatin Research Articles

Formulation, in vitro evaluation and kinetic analysis of chitosan–gelatin bilayer muco-adhesive buccal patches of insulin nanoparticles

The present study was performed to optimise the formulation of a muco-adhesive buccal patch for insulin nanoparticles (NPs) delivery. Insulin NPs were synthesised by an ionic gelation technique using N-di methyl ethyl chitosan cysteine (DMEC-Cys) as permeation enhancer biopolymer, tripolyphosphate (TPP) and insulin. Buccal patches were developed by solvent-casting technique using chitosan and gelatine as muco-adhesive polymers. Optimised patches were embedded with 3 mg of insulin-loaded NPs with a homogeneous distribution of NPs in the muco-adhesive matrix, which displayed adequate physico-mechanical properties. The drug release characteristics, release mechanism and kinetics were investigated. Data fitting to Peppas equation with a correlation coefficient indicated that the mechanism of drug release followed an anomalous transport that means drug release was afforded through drug diffusion along with polymer erosion. In vitro drug release, release kinetics, physical and mechanical studies for all patch formulations reflected the ideal characteristics of this buccal patch for the delivery of insulin NPs.

Enhancing mechanical properties of polyelectrolyte complex nanofibers with graphene oxide nanofillers pretreated by polycation

The wide application of polyelectrolyte complex (PEC) nanofibers has been restrained by their unsatisfactory mechanical properties due to high porosity. In this study, we proposed a strategy for improving the mechanical properties of PEC nanofibers using graphene oxide (GO) fillers pretreated by polycation. Chitosan/gelatin (CS/GE) was adopted as the PEC model system. Through the proposed preparation procedure, GO/CS/GE nanofibers can be obtained successfully by electrospinning. Fourier transform infrared spectroscopy (FITR) and differential scanning calorimetry (DSC) results revealed the strong interactions between GO and PEC matrix, which resulted from the synergistic effect of electrostatic attraction and hydrogen bonding. The effective dispersion of GO in CS/GE matrix was verified through TEM and XRD analysis. As demonstrated by the results of tensile testing, mechanical properties of CS/GE can be effectively enhanced with the addition of GO fillers. At 1.5 wt% filler content, GO/CS/GE nanofiber membranes achieved 358% augment of tensile strength, 175% improvement of Young's modulus and 460% boost of toughness compared with neat CS/GE, which were attributed to effective filler-matrix interactions and good dispersion of fillers. Therefore, the inclusion of GO fillers into PEC through the procedure proposed in this study is a promising strategy for mechanical enhancement of PEC nanofibers.

Rapidly and Effectively Improving the Mechanical Properties of Polyelectrolyte Complex Nanofibers through Microwave Treatment

A facile method for improving the mechanical properties of polyelectrolyte complex (PEC) nanofibers by microwave treatment is proposed and investigated in chitosan/gelatin (CG) PEC model system. The microwave treatment with 600 W for 120 s produces 1.3‐ and 2.6‐folds elevation on Young's modulus and tensile strength of nanofiber membranes, respectively, which is attributed to the enhanced intermolecular interactions and the increased crystallinity in CG composites. In addition, the microwave‐treated CG nanofibers are found still possessing excellent swelling and antibacterial capability, which are indispensable for wound dressing application. Therefore, the microwave treatment can be employed as an effective and rapid strategy to produce PEC nanofibers with ideal mechanical property.

In Vitro and In Vivo Evaluation of Tetracycline Loaded Chitosan-Gelatin Nanosphere Coatings for Titanium Surface Functionalization.

Owing to the biocompatibility of titanium surface, titanium implants are suitable substrates for microbial colonization and biofilm formation, which is still a serious clinical threat. Current research trends have been focused on the development of antibacterial coatings on titanium substrate or adhesion resistant surface. In our previous study, tetracycline (Tc) loaded chitosan-gelatin (CSG) nanosphere coatings are successfully fabricated on titanium substrates via electrophoretic deposition. These coatings show nanosphere structure, and excellent antibacterial property in vitro. However, further in vitro and in vivo evaluation of the coatings is required for the future application. Therefore, in the present study, the authors investigate the coatings' mechanical, swelling and degradation property, in vitro cellular response to preosteoblast cells, and the antibacterial property in rabbits. Results show that Tc incorporation can improve the tensile bond strength of the coating, decrease the swelling ratio, and accelerate the degradation of the coating. Although high Tc concentration group exhibits cytotoxicity to MC3T3-E1 cells, its in vivo antibacterial property is preferred, and shows better outcome than the prophylactic administration of Tc. Tc loaded CSG nanosphere coatings are suitable antibacterial coatings for titanium surface functionalization.

Design and evaluation of buccal films as paediatric dosage form for transmucosal delivery of ondansetron

In the process of implementation and innovation of paediatric dosage forms, buccal films for transmucosal administration of drug represent one of the most interesting approach. In fact, films are able to provide an extended duration of activity allowing minimal dosage and frequency and offer an exact and flexible dose, associated with ease of handling. The objective of the present study was to develop polymeric films for the sustained release of ondansetron hydrochloride, a selective inhibitor of 5-HT3 receptors indicated in paediatrics for the prevention and treatment of nausea and vomiting caused by cytotoxic chemotherapy or radiotherapy and postoperatively. Films were prepared by casting and drying of aqueous solutions containing different weight ratios of hydroxypropylmethylcellulose (HPMC) with chitosan (CH) or sodium hyaluronate (HA) or gelatin (GEL) and characterized for their physico-chemical and functional properties. The presence of HA, GEL and CH did not improve the mucoadhesive properties of HPMC film. The inclusion of GEL and CH in HPMC film increased in vitro drug release with respect to the inclusion of HA, although films containing HA showed the highest water uptake. Moreover in agreement with the release behaviour, the inclusion of CH and GEL provided higher drug permeation through porcine buccal mucosa with respect to HPMC film and ensured linear permeation profiles of drug.

Bactericidal paper trays doped with silver nanoparticles for egg storing applications

In this study, a cost-effective way to deposit the silver nanoparticles (AgNPs) on paper egg trays was developed, which proved suitable for prolonged storage of table eggs for house-hold use without deterioration of egg quality. Silver nanoparticles were synthesized based on chemical reduction approach and mixed with gelatin–chitosan mixer used as a colloidal stabilizer as well as fixing agent. AgNPs-doped paper egg trays were characterized by TEM, SEM, FTIR, EDX and XRD. AgNPs containing egg trays were tested for its bactericidal effect against commonly found bacteria on egg shells, E. coli, S. aureus, Streptococcus spp and Salmonella spp. Storing of eggs in the AgNPs-deposited paper egg trays improved the shelf-life of the eggs by more than 14 days compared to controls (eggs stored in conventional trays). In conclusion, the developed paper trays possessed strong antimicrobial activity and it could be an effective storage material for eggs.

A comparison of cryogel scaffolds to identify an appropriate structure for promoting bone regeneration

To create an ideal graft substitute for regenerating bone, the scaffold should possess osteoconductive, osteoinductive, and osteogenic properties. Hydrogels are a very common scaffold, but the mechanical integrity and nanoporous structure are not advantageous for bone regeneration. Cryogelation is a technique in which the controlled freezing and thawing of a polymer creates a spongy, macroporous structure with ideal structural characteristics and promising mechanical stability. Hydrogels and cryogels of three different materials (chitosan–gelatin, N-vinyl-2-pyrrolidone, and silk fibroin (SF)) were compared to assess the optimal material and form of scaffold for this application. Cryogel and hydrogel structures were tested in parallel to evaluate porosity, swelling, mechanical integrity, cellular infiltration, and mineralization potential. Cryogels proved superior to hydrogels based on swelling potential and mechanical properties. Among the cryogels, SF demonstrated high pore diameter and area, mineralization upon cellular infiltration, and the largest presence of osteocalcin, a marker of bone formation. These results demonstrate the practicality of cryogels for a bone regeneration application and identify SF as a potential material choice.

Surface functionalization with strontium-containing nanocomposite coatings via EPD

Metal orthopedic implants still face challenges in some compromised conditions, partly due to bio-inertness. The present study aimed to functionalize metallic implants with organic-inorganic nanocomposite (strontium-containing chitosan/gelatin) coatings through a simple single-step electrophoretic deposition under mild conditions. The surface characterization and in vitro cellular response were studied and compared with chitosan/gelatin (CS/G) coatings. SEM images suggested the inorganic nanoparticles may be encapsulated within or mixed with organic polymers. The XRD patterns showed that strontium carbonate was generated in the coatings. The TEM images revealed strontium-containing nanoparticles were released from the coatings in PBS. The continuous release after the initial burst release ensured the enduring effects of the functionalized surface. The tensile bond strength of the coatings to the substrates increased after the addition of strontium. In vitro cellular study confirmed that strontium-containing coatings supported the proliferation of MC3T3-E1 cells and exhibited excellent ability to enhance the differentiation of such pre-osteoblasts. Therefore, such organic-inorganic nanocomposite coatings are a promising candidate to functionalize orthopedic implant surfaces.

Biocompatible scaffolds composed of chemically crosslinked chitosan and gelatin for tissue engineering

ABSTRACTChitosan‐based scaffolds are widely studied in tissue regeneration because of their biocompatibility and biodegradability. Scaffolds are obtained by different techniques and can be modified with other polymers allowing controlling their properties. This article discusses the assembling of three‐dimensional chitosan porous scaffolds blended with gelatin. Gelatin was used to enhance cells attachment due to the presence of cell adhesion motifs, while improving mechanical strength. 2,5‐dimethoxy‐2,5‐dihydrofurane (DHF) was used as the crosslinking agent, because it allowed to control the reaction kinetics through temperature, time and DHF concentration. The results indicate that scaffolds morphology, pore sizes and distribution, compressive moduli and biodegradation in vitro with lysozyme, can be customized with variations of gelatin content and crosslinking degree. Scaffolds were neither cytotoxic nor genotoxic for human keratinocytes, exhibiting cell–substrate interactions. Our findings demonstrated that chitosan–gelatin scaffolds crosslinked with DHF, as a new crosslinking agent, are suitable in tissue engineering applications. © 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016, 133, 43814.

Janus emulsion mediated porous scaffold bio-fabrication

A three dimensional biopolymer network structure with incorporated nano-porous calcium phosphate (CaP) balls was fabricated by using gelatin-chitosan (GC) polymer blend and GC stabilized olive/silicone oil Janus emulsions, respectively. The emulsions were freeze-dried, and the oil droplets were washed out in order to prepare porous scaffolds with larger surface area. The morphology, pore size, chemical composition, thermal and swelling behavior was studied by Scanning Electron Microscopy (SEM), Fourier Transform Infrared Spectroscopy (FTIR) and micro-Differential Scanning Calorimetry (micro-DSC). Microscopic analysis confirmed that the pore size of the GC based sponges after freeze-drying may be drastically reduced by using Janus emulsions. Besides, the incorporation of nanoporous calcium phosphate balls is also lowering the pore size and enhancing thermal stability.

Fabrication and characterization of Mg-doped chitosan–gelatin nanocompound coatings for titanium surface functionalization

Titanium and its alloys have been widely used in clinic and achieved great success. Due to the bio-inertness of titanium surface, challenges still exit in some compromised conditions. The present study aimed to functionalize titanium surface with magnesium (Mg)-doped chitosan/gelatin (CS/G) nanocompound coatings via electrophoretic deposition (EPD). CS/G coatings loaded with different amount of magnesium were successfully prepared on titanium substrate via EPD. Physicochemical characterization of the coatings confirmed that magnesium ions were loaded into the coatings in a dose-dependent manner. XRD results demonstrated that co-deposition of magnesium influenced the crystallinity of the coatings, and a new crystalline substance presented, namely hydrated basic magnesium carbonate. Mechanical tests showed improved tensile and shear bond strength of the magnesium-doped coatings, while the excessively high magnesium concentration could eventually decrease the bonding strength. Sustained release of magnesium ion was detected by ICP-OES within 28 days. TEM images also displayed that nanoparticles could be released from the coatings. In vitro cellular response assays demonstrated that the Mg-doped nanocompound coatings could enhance the proliferation and osteogenic differentiation of MC3T3-E1 cells compared to CS/G coatings. Therefore, it could be concluded that Mg-doped CS/G nanocompound coatings were successfully fabricated on titanium substrates via EPD. It would be a promising candidate to functionalize titanium surface with such organic–inorganic nanocompound coatings.

Antimicrobial activity of chitosan coatings and films against Listeria monocytogenes on black radish

The antibacterial activity of chitosan coatings prepared with acetic or lactic acid, as well as of composite chitosan–gelatin films prepared with essential oils, was evaluated in fresh shredded black radish samples inoculated with Listeria monocytogenes ATCC 19115 and L. monocytogenes ATCC 19112 during seven days of storage at 4°C. The chitosan coating prepared with acetic acid showed the most effective antibacterial activity. All tested formulations of chitosan films exhibited strong antimicrobial activity on the growth of L. monocytogenes on black radish, although a higher inhibition of pathogens was achieved at higher concentrations of chitosan. The antimicrobial effect of chitosan films was even more pronounced with the addition of essential oils. Chitosan–gelatin films with thyme essential oils showed the most effective antimicrobial activity. A reduction of 2.4log10CFU/g for L. monocytogenes ATCC 19115 and 2.1log10CFU/g for L. monocytogenes ATCC 19112 was achieved in the presence of 1% chitosan film containing 0.2% of thyme essential oil after 24h of storage.

Fabrication and characterization of novel nano-biocomposite scaffold of chitosan–gelatin–alginate–hydroxyapatite for bone tissue engineering

A novel nano-biocomposite scaffold was fabricated in bead form by applying simple foaming method, using a combination of natural polymers–chitosan, gelatin, alginate and a bioceramic–nano-hydroxyapatite (nHAp). This approach of combining nHAp with natural polymers to fabricate the composite scaffold, can provide good mechanical strength and biological property mimicking natural bone. Environmental scanning electron microscopy (ESEM) images of the nano-biocomposite scaffold revealed the presence of interconnected pores, mostly spread over the whole surface of the scaffold. The nHAp particulates have covered the surface of the composite matrix and made the surface of the scaffold rougher. The scaffold has a porosity of 82% with a mean pore size of 112±19.0μm. Swelling and degradation studies of the scaffold showed that the scaffold possesses excellent properties of hydrophilicity and biodegradability. Short term mechanical testing of the scaffold does not reveal any rupturing after agitation under physiological conditions, which is an indicative of good mechanical stability of the scaffold. In vitro cell culture studies by seeding osteoblast cells over the composite scaffold showed good cell viability, proliferation rate, adhesion and maintenance of osteoblastic phenotype as indicated by MTT assay, ESEM of cell–scaffold construct, histological staining and gene expression studies, respectively. Thus, it could be stated that the nano-biocomposite scaffold of chitosan–gelatin–alginate–nHAp has the paramount importance for applications in bone tissue-engineering in future regenerative therapies.

Improved Biodegradable Radiation Cured Polymeric Film Prepared from Chitosan-Gelatin Blend

The mechanical, thermal, swelling, and release properties of chitosan-gelatin (CG) films have been investigated in order to verify the influence of UV and gamma radiation on the stability of the films. Thin films of chitosan and gelatin (1 : 3, w/w) that were radiated with 100 krad of gamma dose showed the best performance and the TS values reached 25, 45, and 49 MPa, respectively, for chitosan, gelatin, and blend. The corresponding highest TS values were 23, 42, and 45 MPa, respectively, for 10 passes of UV radiation. The effect of radiation over gelatin, chitosan, and CG blend caused modification in the arrangement of molecules in the crystal lattice that is significant by XRD analysis. Surfaces of the films were also investigated by scanning electron microscope (SEM). Fourier transform infrared spectroscopy (FTIR) studies further revealed structural changes of the films. These changes were attributed to understanding the behavior of the irradiated chitosan, gelatin, and CG blend on application of thermal energy using DSC and TGA studies, water uptake of the films in aqueous medium, and soil degradation properties to observe the best possibility for its application.

Electrophoretic deposition of chitosan/gelatin coatings with controlled porous surface topography to enhance initial osteoblast adhesive responses.

Electrophoretically deposited (EPD) coatings have often been employed recently for the addition of different new chemical compositions to classic chitosan coatings to improve the biocompatibility and therapeutic potential of coated implants. However, little attention has been paid to enhance the cell response to EPD coatings via integrating the effects of chemical components and surface topography. Here, we fabricated EPD chitosan/gelatin (CS/G) coatings with controlled porous surface topography by controlling bubble generation in the EPD process via changing the gelatin content in solution from 0, 0.01, 0.1, and 1 to 10 mg ml-1. The pure chitosan coating surface was characterized by homogeneous large pores of 500 μm. After 0.01 mg ml-1 gelatin was added, 180 μm small pores appeared on the walls of large pores. As the gelatin content increased to 0.1 mg ml-1, a number of small pores increased noticeably. When the gelatin content reached 1 mg ml-1, large pores disappeared, and the coating displayed homogeneous small pores. 10 mg ml-1 gelatin concentration led to coatings consisting of small pores with not integral and continuous structures. The initial osteoblastic responses, including cell adherence progress, spreading area, proliferation rate, and focal adhesion-related gene expression, gradually improved from 0 to 0.01, 0.1, and 1 mg ml-1 gelatin content, but decreased from 1 to 10 mg ml-1. All these results indicated that the initial cell responses to coatings reached a peak when it was 1 mg ml-1 gelatin and they had homogeneous small pores, which might contribute to the synergistic effects of the porous surface structure and components. This work would be beneficial for expanding the potential application of EPD coatings.

Application of chitosan films in the quality control of fresh shredded vegetables

The objective of this study was to investigate the antimicrobial effects of chitosan gelatin films with or without added essential oils against Listeria monocytogenes on fresh shredded cabbage, carrot and black radish. Samples of fresh shredded vegetables were inoculated with L. monocytogenes and then stored between two layers of edible chitosan films at various concentrations, with or without added essential oils. The inoculated, film treated samples were stored at 4 °C. Samples were taken every day,during six days, for enumeration of surviving L. monocytogenes by plating on appropriate media. Compared to control,1% chitosan film and 0.5% film-induced to 4.7 and 2.9 log CFU/g reductions on shredded cabbage 3rd day respectively. After 3 days, films with 1% chitosan reduced L. monocytogenes population by 3 log CFU/g on carrots. Antimicrobial activity of chitosan film increases with increasing concentrations of applied chitosan, where 1% chitosan film has the highest antimicrobial activity compared to 0.5% chitosan film. Nevertheless, chitosan films were more effective on black radish than on cabbage and carrot. Addition of thyme essential oils, significantly improves the activity of the antimicrobial film and reduce the time required for the inhibition of bacteria in vegetables. Also, changes of color, taste, aroma and chemical composition of non-treated and treated samples were determined. The application of chitosan films exerts a significant influence on the pH changes and content of soluble solids of stored vegetables. Chitosan films improved sample appearance, since they diminished the occurrence of the color changes during storage. The results of this experiment showed that the use of chitosan films is a viable alternative in controlling the microorganisms present in minimally processed vegetables, improving its sensory quality. The advantages of edible films were the simple manufacturing process, biodegradability and usage of natural and safe compounds.

Composite microsphere-functionalized scaffold for the controlled release of small molecules in tissue engineering

Current tissue engineering strategies focus on restoring damaged tissue architectures using biologically active scaffolds. The ideal scaffold would mimic the extracellular matrix of any tissue of interest, promoting cell proliferation and de novo extracellular matrix deposition. A plethora of techniques have been evaluated to engineer scaffolds for the controlled and targeted release of bioactive molecules to provide a functional structure for tissue growth and remodeling, as well as enhance recruitment and proliferation of autologous cells within the implant. Recently, novel approaches using small molecules, instead of growth factors, have been exploited to regulate tissue regeneration. The use of small synthetic molecules could be very advantageous because of their stability, tunability, and low cost. Herein, we propose a chitosan–gelatin scaffold functionalized with composite microspheres consisting of mesoporous silicon microparticles and poly(dl-lactic-co-glycolic acid) for the controlled release of sphingosine-1-phospate, a small molecule of interest. We characterized the platform with scanning electron microscopy, Fourier transform infrared spectroscopy, and confocal microscopy. Finally, the biocompatibility of this multiscale system was analyzed by culturing human mesenchymal stem cells onto the scaffold. The presented strategy establishes the basis of a versatile scaffold for the controlled release of small molecules and for culturing mesenchymal stem cells for regenerative medicine applications.

Surface functionalization of titanium with tetracycline loaded chitosan–gelatin nanosphere coatings via EPD: fabrication, characterization and mechanism

Tetracycline loaded chitosan–gelatin nanosphere coating has been fabricated on titanium substrate via electrophoretic deposition.

Release of coumarin incorporated into chitosan-gelatin irradiated films

Chitosan and fish gelatin were used to formulate active biobased films containing an antioxidant (coumarin). After drying, the films were irradiated at 40 and 60 kGy using an electron beam accelerator. The effect of irradiation on the film properties as well as the coumarin release mechanism were investigated and compared with the control. Electron Spin Resonance (ESR) revealed free radical formation during irradiation in films containing coumarin. Antioxidant addition and/or irradiation treatment at a dose of 60 kGy resulted in a shift of amide A and amide B peaks. Furthermore a shift of amide II band was only observed for the control film at the same dose.Irradiation allowed improving the thermal stability of the control films. Both irradiation process and addition of coumarin increased the surface wettability (increase of the polar component of the surface tension). From the water barrier analysis, neither irradiation nor coumarin addition influenced the permeability at the lower RH gradient used (0–30% RH). Using the higher RH gradient (30–84%) induced a rise of the WVP of all films (containing or not coumarin) after irradiation treatment. At 60 kGy, the tensile strength of only the control films increased significantly. Considering coumarin release from the film in aqueous medium, the apparent diffusion coefficient of coumarin is two times reduced after irradiation. Irradiation also allowed to better protect the incorporated antioxidant. Indeed, the amount of coumarin in the non-irradiated film was significantly lowered compared to the initial quantity, which is probably due to chemical reactivity.

Healing of skin wounds with a chitosan–gelatin sponge loaded with tannins and platelet-rich plasma

A chitosan–gelatin sponge (CSGT) was prepared using a chitosan/ascorbic acid solution blend containing gelatin, followed by crosslinking with tannin acid and freeze-drying, thereby combining the chitosan sponge and gelatin sponge. The structure of the CSGT was observed by scanning electron microscopy and was shown to have uniform and abundant pores measuring about 145–240μm in size. We also characterized the sponges by infrared spectroscopy, thermogravimetric analysis, mechanical property tests, swelling behavior analysis, water retention capacity tests, antibacterial property analysis, and cytotoxicity tests. Our data showed that the CSGT had good thermostability and mechanical properties as well as efficient water absorption and retention capacities. Moreover, the CSGT could effectively inhibit the growth of Escherichia coli and Staphylococcus aureus with low toxicity. In animal experiments, macroscopic observations and histological examinations showed that the wound covered by the CSGT healed quickly. Additionally, loading of the CSGT with platelet-rich plasma resulted in further acceleration of wound healing. Therefore, the CSGT and the CSGT with platelet-rich plasma were suitable for application as a wound dressing and may have potential for use in various biomedical applications.