HISTOMORPHOLOGY AND PHYSICO-CHEMICAL EVALUATION OF RAW AND DECELLULARISED TUNICA VAGINALIS OF BUFFALO FOR USE AS A BIOMATERIAL IN REGENERATIVE MEDICINE
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Date
2021-09-30
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COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR
Abstract
The 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.
Description
Submitted in partial fulfilment of the requirement for the degree of
Master of Veterinary Science of Veterinary Anatomy and Histology