STRUCTURAL AND PHYSICO-CHEMICAL EVALUATION OF RAW AND DECELLULARISED PORCINE TUNICA VAGINALIS AS A NOVEL ACELLULAR MATRIX
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Date
2021-12-30
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COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR
Abstract
The current research was aimed to develop a novel acellular matrix from
porcine tunica vaginalis by a standardised decellularisation protocol and to study the
structural and physico-chemical properties of raw tunica vaginalis and the developed
acellular matrix. Due to the intimate association of visceral layer of tunica vaginalis
with testicular capsule, parietal layer of tunica vaginalis (PTV) were collected for the
development of the acellular matrix from the testes of six adult healthy pigs slaughtered
at the Meat Technology Unit, College of Veterinary and Animal Sciences, Mannuthy.
Grossly PTV was found as an extensive, thin, elastic, flask shaped serous sac with
numerous blood vessels and nerves. It lined the scrotum and the inguinal canal and was
attached to the tail of epididymis and vas deferens. Histologically PTV comprised
mesothelium and dense regular connective tissue composed of thick collagen bundles,
a few elastic, reticular and smooth muscle fibres, blood vessels, and nerves. PTV was
decellularised by two protocols using aqueous solution of bovine bile and Trypsin EDTA. After decellularisation, physico-chemical properties of the acellular matrix
were evaluated and compared with that of raw PTV. Thickness, tensile strength,
collagen content and resistance to collagenase enzyme of the decellularised matrix
were less when compared with the raw PTV, whereas the collagen solubility was
higher. Efficiency of decellularisation was assessed by routine and special histological
staining methods, nuclear staining using 4, 6-Diamidino-2-phenylindole dihydro
chloride, estimation of glycosaminoglycans, analysis of DNA content and by scanning
electron microscopy. Lack of cellularity, lowest DNA content (<50 ng/µL) and
increased porosity of acellular matrix revealed that efficiency of decellularisation using
Trypsin-EDTA was better than that with bile treatment.
Description
Submitted in partial fulfilment of the requirement for the degree of
MASTER OF VETERINARY SCIENCE in Veterinary Anatomy and Histology