Indu V RajLHINGNEIVAH DOUNGEL2022-11-012022-11-012021-09-30https://krishikosh.egranth.ac.in/handle/1/5810189489Submitted in partial fulfilment of the requirement for the degree of Master of Veterinary Science of Veterinary Anatomy and HistologyThe present study was undertaken to develop an acellular matrix from buffalo bull tunica vaginalis by a newly standardised decellularisation technique. Ten pairs of testes were collected hygienically from ten healthy adult Murrah buffalo bulls slaughtered at the Meat Technology Unit, College of Veterinary and Animal Sciences, Mannuthy. For decellularization, the fresh parietal layer of tunica vaginalis removed from the testes samples were processed as per the newly standardised protocol using trypsin-EDTA with triton-X-100 as detergent. The histomorphological, ultrastructural and physico-chemical characteristics of the raw and decellularised tunica vaginalis were compared and efficiency of decellularisation was evaluated by histological staining techniques, total DNA quantification and scanning electron microscopy. It was observed that the average weight of the left testis in adult Murrah buffalo bulls with and without parietal layer of tunica vaginalis (PTV) was more than that of the right testis and exhibited a statistically significant difference. The outer parietal layer of tunica vaginalis that lined the scrotum could easily be separated out from the testis. The vertical span of PTV of left testis was more than that of the right and the difference was statistically significant. Histologically the PTV consisted of a thin mesothelium and dense regular connective tissue made up of thick collagen bundles, a few elastic fibres and reticular fibres supported by blood vessels, smooth muscle fibres and nerves. It was observed that the decellularised PTV were devoid of cells and genetic materials such as DNA while maintaining its overall tissue histoarchitecture. The physico-chemical characteristics like mean thickness, tensile strength and collagen concentration of decellularised buffalo PTV samples were significantly reduced while the collagen solubility per cent and percentage weight loss after collagenase degradation were significantly higher in decellularised PTV than raw PTV samples. It was concluded that the samples of parietal layer of tunica vaginalis of buffalo prepared by using the newly standardised decellularisation technique showed acceptable physico-chemical properties required for a biologic scaffold. It may be used as a biomaterial in regenerative medicine after extensive clinical screening.EnglishThesis