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Subject: Animal Biochemistry × Author: HAZORIKA, MOUSUMI × End date: 2017 × Start date: 2015 × Thesis Type: Ph.D ×
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    ThesisItemOpen Access
    THERAPEUTIC POTENTIAL OF BONE-MARROW DERIVED MESENCHYMAL STEM CELLS IN WOUND HEALING
    (Assam Agricultural University, Khanapara, Guwahati, 2017-07) HAZORIKA, MOUSUMI; Kalita, Dhruba Jyoti
    The present experiment was carried out to explore the therapeutic potential of rabbit bone-marrow derived mesenchymal stem cells (rBM-MSCs) in wound healing in New Zealand White rabbits. The experiment was conducted in the Department of Veterinary Biochemistry, College of Veterinary Science, Assam Agricultural University, Khanapara, Guwahati-781022. Eighteen numbers of clinically healthy male New Zealand white rabbit of 3-4 months with mean body weight of 1.52 kg were randomly divided into three different groups of six animals each viz. Gr-I (Control group), Gr-II (Test group-1) and Gr-III (Test group-2). A total of thirty-six surgical wounds were created on the skin of eighteen rabbits with two in each. The Gr-I rabbits received no treatment and was considered as Control group. Gr-II rabbits were treated with a standard drug i.e. Povidine iodine ointment (Betadine®, USP 10 % w/w) while Gr–III rabbits were treated with cultured BM-MSCs topically. They were raised under standard laboratory conditions and veterinary supervision throughout the experimental period and were given standard pellet feed. The experiment was conducted in a 2x2 factorial design for a period of 30 days. Bone marrow was collected from rabbits to culture and characterize the BM-MSCs. Blood was collected from all the animals of each group on days 0, 3, 7, 15 and 30 to estimate haemato-biochemical constituents viz. haemoglobin, PCV, ESR, TEC, TLC and DLC; serum glucose, total protein, triglycerides, SGOT, SGPT, alkaline phosphatase, uric acid, Ca2+, P, Mg2+, Na+, K+, Cl-, iron, copper, zinc, cytokine proteins: TNF-α, IL-1β, IL-4, IL-6, IL-10 and to study cytokine gene expression pattern during wound healing. Wound area was measured on days 1, 3, 7, 15, 21 and 30 of each animal, of different experimental groups. Tissue was collected on day 10 and day 20 of surgery from each animal of different experimental groups to estimate certain granulation tissue markers viz. LPO, NO; SOD, CAT, GSH, vitamin A, vitamin C, vitamin E; MPO; protein and DNA; and connective tissue markers viz. HYP, HXA, HUA, collagen and elastin. The tissue was also collected on days 1, 10 and 20 of surgery to observe the histopathological changes during wound healing. The rBM-MSCs was isolated, cultured and characterized as per the standards set by ISCT. The rBM-MSC treated group showed complete healing of wound by day 21 whereas complete healing of wound was observed on day 30 in both Gr-I and Gr-II. All the haemato-biochemical constituents were found within the normal range throughout the experimental period. However, the levels of certain parameters significantly increase or decrease amongst the groups with the advancement of time. The rBM-MSCs significantly increased the antioxidants’ levels and reduced the generation of free radicals’ and acute inflammatory marker on day 10 and day 20 of surgery. Significant increase observed in the content of granulation tissue biochemical markers (protein and DNA) and connective tissue markers (HYP, HXA, HUA, collagen and elastin). Histopathologically, the BM-MSCs treated group showed early tissue changes compared to other groups and approximately resembles the adjacent normal skin on day 20. On the basis of macroscopic, biochemical and histopathological evaluation of wounds, it can be concluded that the rBM-MSCs can be used successfully for the treatment of surgical wounds without any significant adverse effects on the animal.