Abstract
In the present study, protocols for decellularizing the buffalo rumen were optimized. The rumen was made acellular using ionic detergents (sodium deoxycholate), nonionic detergents (Triton X-100 and Tween 20), zwitterionic detergents (tri (n-butyl) phosphate), and enzymes (trypsin) in 0.5% and 1% concentration. Treatment with detergents was done up to 96 h, and the solution was changed at every 12 h time intervals, and samples were collected at 12, 24, 48, 72, and 96 h time intervals for macroscopic and microscopic observations, DNA quantification, and in vitro cytocompatibility analysis. All the specimens treated for different time intervals were adequately decellularized, except enzyme treatment. Treatment with trypsin enzyme was inefficient in decellularization during various time intervals. Complete acellularization was achieved with 1% sodium deoxycholate at 72 h. Masson’s trichrome staining revealed thick, transversely and longitudinally arranged collagen fibers. The prepared acellular rumen matrices were successfully tested in 26 buffalo calves, aged between 3 and 27 months and weighing 50–115 kg, suffering from umbilical hernia with an average diameter 6–12.5 cm of the hernial ring.
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Singh, A.K. et al. (2022). Tissue Scaffolds Derived from Buffalo Rumen and Clinical Applications. In: Kumar, N., Kumar, V., Shrivastava, S., Gangwar, A.K., Saxena, S. (eds) Tissue Scaffolds. Springer Protocols Handbooks. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2425-8_13
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DOI: https://doi.org/10.1007/978-1-0716-2425-8_13
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