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Kerala Veterinary and Animal Sciences University, Wayanad

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20211
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Subject: Veterinary Anatomy and Histology × Author: ABHIN RAJ K.P. × Start date: 2020 ×
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    ThesisItemOpen Access
    STRUCTURAL AND PHYSICO-CHEMICAL EVALUATION OF RAW AND DECELLULARISED PORCINE TUNICA VAGINALIS AS A NOVEL ACELLULAR MATRIX
    (COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, 2021-12-30) ABHIN RAJ K.P.; Sreeranjini A.R
    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.