Comparative evaluation and optimization of amniotic membrane derived bioscaffold for bone regeneration in critical sized bone defect in rabbit radius model.

Abstract

Background: Bone regeneration and repair are critical research areas within regenerative medicine, aiming to address the challenges posed by critical-sized bone defects. Bioscaffolds and cell-based therapies have been explored to enhance osteogenesis and promote effective bone regeneration. This study aimed to assess the regenerative potential of rabbit amniotic membrane (rAM) derived bioscaffold and its comparative evaluation with another bioscaffold, the decellularized periosteum (DP) of the buffalo rib, which was recellularized with rAM derived mesenchymal stem cells (MSCs).

Methods: Passage 3 (P3) rAM-MSCs were validated for positive and negative stemness marker expression by PCR, immunolocalization and trilineage differentiation. Fresh rAM was cryopreserved for three months. An autologous rabbit model was used to assess the osteogenic capacity of different bioscaffolds: fresh rAM (Group II), frozen-thawed rAM (Group III), DP (Group IV), and DP enriched with rAM-MSCs (Group V) and control (Group I) in critical-size defect of 10 mm in radius bone. Radiographic evaluation was performed on the 1^st, 60^th and 90^th days, and ultramicroscopic and histomorphological evaluations were performed.

Results: Groups II and V presented the highest levels of remodeling and osteogenesis, a reduction in the defect size and total radiological scores. Groups II and V had the highest levels of osteogenesis, bone marrow development, cortical bone production, and medullary bone formation and the highest total histology score.

Conclusion: These findings revealed that fresh rAM, a rich source of MSCs, and DP enhanced with rAM-MSCs are the preferred bioscaffolds for critical-sized bone defect repair.