Rejoining of cut wounds by engineered gelatin-keratin glue.

BACKGROUND: Rejoining of cut tissue ends of a critical site challenges clinicians. The toxicity, antigenicity, low adhesive strength, flexibility, swelling and cost of the currently employed glue demands an alternative. Engineered gelatin-keratin glue (EGK-glue) described in the present study was found to be suitable for wet tissue approximation. METHODS: EGK-glue was prepared by engineering gelatin with caffeic acid using EDC and conjugating with keratin by periodate oxidation. UV-visible, (1)H NMR and circular dichroism analyses followed by experiments on gelation time, rheology, gel adhesive strength (in vitro), wet tissue approximation (in vivo), H&E staining of tissue sections at scheduled time intervals and tensile strength of the healed skin were carried out to assess the effectiveness of the EGK-glue in comparison with fibrin glue and cyanoacrylate. RESULTS: Results of UV-visible, NMR and CD analyses confirmed the functionalization and secondary structural changes. Increasing concentration of keratin reduces the gelation time (<15s). Lap-shear test demonstrates the maximum adhesive strength of 16.6±1.2kPa. Results of hemocompatibility and cytocompatibility studies suggested the suitability of the glue for clinical applications. Tissue approximation property assessed using the incision wound model (Wistar strain) in comparison with cyanoacrylate and fibrin glue suggested, that EGK-glue explicitly accelerates the rejoining of tissue with a 1.86 fold increase in skin tensile strength after healing. CONCLUSIONS: Imparting quinone moiety to gelatin-keratin conjugates through caffeic acid and a weaker oxidizing agent provides an adhesive glue with appreciable strength, and hemocompatible, cytocompatible and biodegradable properties, which, rejoin the cut tissue ends effectively. GENERAL SIGNIFICANCE: EGK-glue obtained in the present study finds wide biomedical/clinical applications.

Engineering of chitosan and collagen macromolecules using sebacic acid for clinical applications.

Transformation of natural polymers to three-dimensional (3D) scaffolds for biomedical applications faces a number of challenges, viz., solubility, stability (mechanical and thermal), strength, biocompatibility, and biodegradability. Hence, intensive research on suitable agents to provide the requisite properties has been initiated at the global level. In the present study, an attempt was made to engineer chitosan and collagen macromolecules using sebacic acid, and further evaluation of the mechanical stability and biocompatible property of the engineered scaffold material was done. A 3D scaffold material was prepared using chitosan at 1.0% (w/v) and sebacic acid at 0.2% (w/v); similarly, collagen at 0.5% (w/v) and sebacic acid at 0.2% (w/v) were prepared individually by freeze-drying technique. Analysis revealed that the engineered scaffolds displayed an appreciable mechanical strength and, in addition, were found to be biocompatible to NIH 3T3 fibroblast cells. Studies on the chemistry behind the interaction and the characteristics of the cross-linked scaffold materials suggested that non-covalent interactions play a major role in deciding the property of the said polymer materials. The prepared scaffold was suitable for tissue engineering application as a wound dressing material.

Preparation and characterization of malonic acid cross-linked chitosan and collagen 3D scaffolds: an approach on non-covalent interactions.

The present study emphasizes the influence of non-covalent interactions on the mechanical and thermal properties of the scaffolds of chitosan/collagen origin. Malonic acid (MA), a bifuncitonal diacid was chosen to offer non-covalent cross-linking. Three dimensional scaffolds was prepared using chitosan at 1.0% (w/v) and MA at 0.2% (w/v), similarly collagen 0.5% (w/v) and MA 0.2% (w/v) and characterized. Results on FT-IR, TGA, DSC, SEM and mechanical properties (tensile strength, stiffness, Young's modulus, etc.) assessment demonstrated the existence of non-covalent interaction between MA and chitosan/collagen, which offered flexibility and high strength to the scaffolds suitable for tissue engineering research. Studies using NIH 3T3 fibroblast cells suggested biocompatibility nature of the scaffolds. Docking simulation study further supports the intermolecular hydrogen bonding interactions between MA and chitosan/collagen.

Bonding interactions and stability assessment of biopolymer material prepared using type III collagen of avian intestine and anionic polysaccharides.

The present study demonstrate bonding interactions between anionic polysaccharides, alginic acid (AA) and type III collagen extracted from avian intestine used for the preparation of thermally stable and biodegradable biopolymer material. Further the study describes, optimum conditions (pH, temperature and NaCl concentration) required for the formation of fibrils in type III collagen, assessment on degree of cross-linking, nature of bonding patterns, biocompatibility and biodegradability of the cross-linked biomaterial. Results revealed, the resultant biopolymer material exhibit high thermal stability with 5-6 fold increase in tensile strength compared to the plain AA and collagen materials. The degree of cross-linking was calculated as 75%. No cytotoxicity was observed for the cross-linked biopolymer material when tested with skin fibroblast cells and the material was biodegradable when treated with enzyme collagenase. With reference to bonding pattern analysis we found, AA cross-linked with type III collagen via (i) formation of covalent amide linkage between -COOH group of AA and ε-NH2 group of type-III collagen as well as (ii) intermolecular multiple hydrogen bonding between alginic acid -OH group with various amino acid functional group of type-III collagen. Comparisons were made with other cross-linking agents also. For better understanding of bonding pattern, bioinformatics analysis was carried out and discussed in detail. The results of the study emphasize, AA acts as a suitable natural cross-linker for the preparation of wound dressing biopolymer material using collagen. The tensile strength and the thermal stability further added value to the resultant biopolymer.

Preparation and characterization of a thermostable and biodegradable biopolymers using natural cross-linker.

The present study describes preparation and characterization of a thermally stable and biodegradable biopolymer using collagen and a natural polymer, alginic acid (AA). Required concentration of alginic acid and collagen was optimized and the resulting biopolymer was characterized for, degree of cross-linking, mechanical strength, thermal stability, biocompatibility (toxicity) and biodegradability. Results reveal, the degree of cross-linking of alginic acid (at 1.5% concentration) with collagen was calculated as 75%, whereas it was 83% with standard cross-linking agent, glutaraldehyde (at 1.5% concentration). The AA cross-linked biopolymer was stable up to 245°C and Exhibits 5-6-fold increase in mechanical (tensile) strength compared to plain collagen (native) materials. However, glutaraldehyde cross-linked material exhibits comparatively less thermal stability and brittle in nature (low tensile strength). With regard to cell toxicity, no cytotoxicity was observed for AA cross-linked material when tested with mesenchymal cells and found degradable when treated with collagenase enzyme. The nature of bonding pattern and the reason for thermal stability of AA cross-linked collagen biopolymer was discussed in detail with the help of bioinformatics. A supplementary file on efficacy of AACC as a wound dressing material is demonstrated in detail with animal model studies.

Biopolymer from microbial assisted in situ hydrolysis of triglycerides and dimerization of fatty acids.

The present study demonstrates biopolymer production by in situ bio-based dimerization of fatty acids by microorganism isolated from marine sediments. Microbial isolate grown in Zobell medium in the presence of triglycerides for the period of 24-240 h at 37 degrees C, hydrolyze the applied triglycerides and sequentially dimerized the hydrolyzed products and subsequently polymerized and transformed to a biopolymer having appreciable adhesive properties. Physical (nature, odour, stickyness and tensile strength), chemical (instrumentation) and biochemical (cell free broth) methods of analyses carried out provided the hypotheses involved in the formation of the product as well as the nature of the product formed. Results revealed, lipolytic enzymes released during initial period of growth and the biosurfactant production during later period, respectively, hydrolyze the applied triglycerides and initiate the dimerization and further accelerated when the incubation period extended. The existence and the non-existence of in situ hydrolysis of various triglycerides followed by dimerization and polymerization and the mechanism of transformation of triglycerides to biopolymer are discussed in detail.