Allogeneic Amniotic Membrane-Derived 3-Dimensional Culture of Mesenchymal Stem Cells Promotes Rapid Burn Wound Closure in a Non-Human Primate Burn Wound Injury Model

Abstract

Background: Amniotic membrane is a laboratory medical waste that potentially serves as a good source of mesenchymal stem cell (AM-MSCs). This study is aimed to investigate the feasibility and safety of allogeneic three-dimensional (3D) micro-carrier cultures of AM-MSCs transplantation that promotes rapid closure of the third-degree burns wound using non-human primate (NHP), Macaca fascicularis, burn wound injury model. Methods: AM-MSCs was isolated by standard protocol in two-dimensional (2D) cultures following the increasing process of cells in 3D micro-carrier cultures. These were analyzed for its morphology, ability to proliferate, surface markers and multilineage differentiation potential. The 3D AM-MSCs were then injected under the burns wound lesion of the NHP burn wound injury model. Then, we observed the growth factors, the speed of healing process and the scars formation of 3D AM-MSCs, compared to the gold standard treatment of silver sulfadiazine (Burnazine) ointment. Samples treated with the physiological sodium chloride (NaCl 0,9%) served as a negative control. Results: The surface antigen phenotype, morphology and differentiation characteristics of 3D AM-MSCs exhibited properties of MSCs. The speed of wound closure was significantly accelerated when 3D AM-MSCs were applied in NHP burn wound injury model compared to the group with topical application of Burnazine ointment. HGF and VEGF levels were increased, and there were no scars formation on the lesions treated with 3D AM-MSCs. The healing speed were quickly restored, and we found new hair growth in the lesion area found in 3D AM-MSCs-group. Conclusion: 3D AM-MSCs transplantation effectively promotes rapid wound closure in NHP burn wound model. The large amount of cells requirement are solved by 3D cultures that will facilitate the development and improve the burn treatment in clinical set-up. Funding: R.N. supported by the Research and Development of PT Bio Farma (Persero), Life Scinces Company, Bandung, Indonesia, 2018/2019. A.F. supported by the Grants-in-Aid from the Ministry of Research, Technology and Higher Education of the Republic of Indonesia 06/E/KPT/2019 (World Class Research). Declaration of Interest: Authors have declared that no competing interests exist. Ethical Approval: This study was approved by the Ethics Committee of the Institutional of Animal Care and Use Ethics Commission (IACUC) of PT Bio Farma (Persero) No. 01/IACUC-BF/III/2017 with a validity period of 2 years, April 14th, 2017 2019.