The present study aims at the preparation of silk fibroin solution for possible applications in tissue engineering. Pure silk fibroin protein was extracted from Bombyx mori silk cocoon by degumming method using aqueous Na 2 CO 3 solution followed by solubilizing in LiBr aqueous solution. The fibroin protein solution was purified and studied for the protein content. The degumming & solubility were significantly dependent on the salt concentration, treatment temperature, and time. A salt concentration of 0.02 M Na 2 CO 3, temperature of 80°C and time more than 40 min were found to be favorable degumming conditions. The proper conditions of dissolution were found as 9.3 M LiBr, 70 °C temperature, and 3 h dissolving time. The morphology of degummed silk were investigated by SEM at different magnification. SEM revealed the absence of glue like sericine over the silk fibroin surface at optimal degumming condition. The fibroin protein content of the solution was measured by Bradford protein assay. The results indicate that the regenerated silk fibroin (RSF) can be used for fabrication of porous silk fibroin scaffolds for various tissue engineering applications.
In this study, a porous silk fibroin (SF) scaffold was modified with soluble eggshell membrane protein (SEP) with the aim of improving the cell affinity properties of the scaffold for tissue regeneration. The pore size and porosity of the prepared scaffold were in the ranges 200-300 lm and 85-90%, respectively. The existence of SEP on the scaffold surface and the structural and thermal stability were confirmed by energy-dispersive X-ray spectroscopy, X-ray diffraction, Fourier transform infrared spectroscopy, differential scanning calorimetry, and thermogravimetric analysis. The cell culture study indicated a significant improvement in the cell adhesion and proliferation of mesenchymal stem cells (MSCs) on the SF scaffold modified with SEP. The cytocompatibility of the SEP-conjugated SF scaffold was confirmed by a 3-(4,5-dimethyltriazol-2-y1)-2,5-diphenyl tetrazolium assay. Thus, this study demonstrated that the biomimic properties of the scaffold could be enhanced by surface modification with SEP. V C 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014, 131, 40138.
Optimization of protein extraction using silk cocoon was carried out by the design of experiment (DOE) to obtain the response surface methodology ( RSM ). Box-Behnken rotatable design was used to create an experimental program to provide data to model the effects of various factors on protein extraction efficiency. The variables chosen were sodium carbonate concentration ( 1 X ), Lithum bromide concentration ( 2 X ) and temperature ( 3X ). The mathematical relationship between protein extraction efficiency and three significant independent variables can be approximated by a second order quadratic model. RSM was used to describe the individual and interactive effects of three variables at three levels, combined according to a Box-Behnken Design. The coefficient of determination ( 2R ) for the model is 0.9761. Probability value ( P < 0.0001) demonstrates a very high significance for the regression model indicating that Box-Behnken Design can be applied to the protein extraction from silk cocoon, and it is an economical way of obtaining the maximum amount of information with the fewest experiments.
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