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In the present study, fibroblasts seeded matrices prepared from bovine pericardium was evaluated in the repair of full-thickness skin defects in rats. The pericardium tissue from buffalo was treated with 1% sodium dodecyl sulphate (SDS) for 48 hours showed complete acellularity with normal thickness and arrangements of collagen fibres. In-vitro primary mouse embryonic fibroblasts (p-MEF) cells were cultured and seeded on the acellular pericardium matrix (APM) using static seeding technique. Forty-eight (48) rats of either sex were randomly divided into 4 groups of 12 animals each. Under general anaesthesia, a 20x20 mm2 full-thickness skin defect was created on dorsal thoracic area. The defect in group A (control) was repaired with standard dressing material whereas, groups B, C and D animals were repaired with autograft, APM and p-MEF seeded APM, respectively. Wound healing was evaluated on the basis of clinical, gross, immunological and histopathological observations. On the basis of different observations recorded it can be said that APM has wound healing potency however, seeding with p-MEF increases the wound healing potency. APM seeded p-MEF was found to be a novel biomaterial which has amphipathic character in full-thickness skin repair and enhances wound healing.