EVALUATION OF IN VIVO HOST RESPONSE OF DIFFERENTLY DECELLULARISED OMENTAL SCAFFOLDS
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Date
2019-12-31
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COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR
Abstract
In this study, efficiency of decellularisation protocols as well as host
response of differently decellularised bovine omental scaffolds were evaluated
in a rat model. Bile treated omental scaffold (BBO), triton X- 100 treated
omental scaffolds (TBO) and un-processed omental scaffolds (UBO) were used
during the implantation period along with sham control (SCO). Efficiency of
different decellularisation protocols was assessed by DNA quantification, 4, 6-
diamidino-2-phenylindole (DAPI) staining, scanning electron microscopy and
histological examination. Cellular and tissue response was evaluated semi quantitatively using ISO-10993-6: 2016 at 7, 14 and 30 days post- implantation
(PI). CD4+ and CD8+ T-cells in retrieved implants and spleen were evaluated
immunohistochemically to study the graft acceptance of these implants.
Proliferation index was also evaluated by Ki67 immunolabelling. The results of
this study showed that bile treatment had better decellularisation efficiency
than triton X- 100. BBO showed better remodeling response in terms of
neovascularisation, fatty infiltration and neocollagenisation than TBO and
UBO. At all time periods studied, BBO implant had significantly (p< 0.05)
lesser CD4+ per cent and higher CD8+ per cent than TBO and UBO,
respectively. UBO showed significantly higher CD4+ per cent at all time
periods than BBO and TBO. The CD4+/CD8+ ratio for BBO was significantly
(p< 0.05) lower than TBO and UBO. Besides, BBO showed high Ki67 index
(p<0.05) than TBO at 30 days PI. These results document that bile as an
effective decellularising agent with less tissue toxicity than chemical detergent,
triton X- 100. Apart from these, BBO enhanced remodeling potential through
neovascularisation, cellular proliferation and neocollagenisation with
significantly lower CD4+/CD8+ ratio and higher Ki67 index. Thus, bile treated
omental scaffold may be a promising candidate for applications in human and
veterinary medicine for improving patient’s life.
Description
Thesis Submitted in partial fulfillment of the requirement for the degree of Master of Veterinary science in Veterinary Pathology