Keratin from the hoof is a less explored source for making valuable products. In this paper we present the extraction of pure keratin from bovine hooves and characterized them to better address the possible exploitation of this bio-resource as an alternative material for tissue engineering applications. The keratin protein from the pulverized hooves was extracted by reduction, which was observed to be pure, and two polypeptide chains of molecular weight in the range of 45–50 and 55–60 KDa were determined using SDS-PAGE assay. FTIR analysis complementing circular dichroism (CD) data, established that hoof keratin predominantly adopted α-helical conformation with admixture of β-sheet. The keratin was shown to have appreciably high denaturation temperature (215°C) as indicated by differential scanning calorimetric (DSC) analysis. Thermogravimetric analysis (TGA) also showed the retention of 50% of the original weight of the sample even at a temperature of 346°C. The keratin from the hoof had been observed to be biocompatible when analyzed with MTT assay using fibroblast cells, showing more than 90% cell viability. Hence, hoof keratin would be useful for high value biomedical applications.

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Fabrication of keratin-collagen (KC) 3D scaffold with improved thermal denaturation rate is reported. In vitro application of (KC) scaffold stimulates basic extra cellular matrix constituents. KC Scaffold considerably reduced undesirable properties of both collagen and keratin while collagen incorporation reduces the fragility with increases of strength and flexibility in the scaffold. In addition to this, the scaffold showed homogenous well-interconnected pores in the range of 10-100 mm when observed in scanning electron microscope. Usage of keratin in KC scaffold offers increased biodegradation rate and higher denaturation rate in addition to its rapid cell growth with normal morphology ultimately reaching cell population of 3.9-9.7 million per cm 3 after 48 hr in KC scaffold. Circular dichroism (CD) and Fourier transform spectroscopy (FT-IR) of KC showed presence of helical structure of collagen and ß-turns of keratin confirming retention of native structures of both the proteins KC scaffold showed good swelling behavior and water uptake. Our study strongly supports the superidity of KC scaffold over the collagen or keratin when they are independently used for tissue engineering applications.