Expression of Growth Associated Protein-43 (GAP-43) and Brain-derived Neurotrophic Factor (BDNF) from Adipose Mesenchymal Stem Cells (ASC) with fresh frozen nerve scaffold in 5% hypoxic condition

Abstract

Background: Peripheral nerve injury is one of the most common musculoskeletal injuries, and the healing is prolonged and affects the patient's quality of life. Secretomes derived from Adipose Stem Cells (ASC) is widely researched in tissue engineering. Various studies currently focus on observing potential scaffolds in nerve regeneration. This study aims to analyze the expression of Growth Associated Protein-43 (GAP-43) and Brain-derived Neurotrophic Factor (BDNF) produced by ASC culture with fresh frozen nerve scaffold under 5% hypoxic conditions in vitro.Methods: An in vitro experimental laboratory research using the randomized control group post-test only design was conducted at the Dr. Soetomo Hospital Tissue Bank, Surabaya, East Java, Indonesia. Isolated mesenchymal stem cells from fat tissue were characterized using CD-105. If more than 90% of the MSCs membrane surface binds to the CD-105 marker is indicated positive. Hypoxic conditions were applied to culture stem cells with oxygen levels of 5% and several days of cultivation time until they reached passage four and passage eight. ELISA-test for GAP-43 and BDNF was carried out using secretomes obtained from ASC culture with fresh frozen nerve scaffold in normoxic (21%) and hypoxic (5%) conditions. The analysis was performed on the 2nd to 4th day and 6th day of culture using IBM® SPSS® program version 23 for Windows.Results: In this study, there was a significant difference in BDNF expression in the group given fresh frozen nerve scaffold (p=0.040) but no significant difference in the expression of GAP-43 in the group given fresh frozen nerve scaffold (p=0.214) in normoxic conditions.Conclusion: The 5% hypoxic medium did not provide a significant difference in the expression of GAP-43 and BDNF.