Osteogenic Potential Differentiation of Human Amnion Mesenchymal Stem Cell with Chitosan-Carbonate Apatite Scaffold (In Vitro Study)

Abstract

Background: Tissue engineering based approaches have received much attention. Incorporation of chitosan and carbonate apatite (CA) improve its capability. Human mesenchymal stem cells (hMSCs) is viable for xenogenic transplantation. The purpose of this study was to fabricate and evaluate the osteogenic potential differentiation of human amnion mesenchymal stem cell with carbonate apatitechitosan scaffolds (CA-ChSs) for tissue engineering. Method: Human amniotic membrane was procured from using cesarean section. Soncini's protocol was employed for the isolation procedure. The cells cultured on collagen-coated dishes using Dulbecco's minimal essential medium (DMEM)/F12 (1:1). A chitosan powder of medium molecular weight deacetylated chitin, poly(D(glucosamine) was used and mixed with CA. Immunocytochemistry and flowcytometry used for phenotypic characterization of hAMSC.Result: Amniotic membrane obtained using cesarean section under aseptic condition did not exhibit any growth of cell cultures which were not contaminated. Immunocytochemistry testing revealed that the target cells expressed strong mesenchymal stem cell marker CD 105. Characterization at passage 10 showed that CD44 was the most significant and abundant surface receptors. The number of viable cells in chitosan-carbonate apatite was 66.59%. Scanning electron microscope (SEM) observation revealed that CA-ChSs had threedimensional structure with many pores and hAMSc could attached and proliferation among the porosity of the scaffold. The formation of calcium in the cell as an indicator of osteoblast cells was detected using Alizarin Red solution. Conclusion: hAMSc harvested from human amniotic membrane seeding in CA-ChSs had the capability for in vitro osteogenesis makes them be the one of the potential options for bone tissue engineering.Keywords: human amniotic membrane mesenchymal stem cells, chitosan-carbonate apatite, scaffold, MSC phenotypic characterization, MTT assay. Cite This Article: Kamadjaja, M., Salim, S., Rantam, F. 2016. Osteogenic potential differentiation of human amnion mesenchymal stem cell with chitosan-carbonate apatite scaffold (in vitro study).

<i>In vitro</i> bone sialoprotein-I expression in combined gingival stromal cells and platelet rich fibrin during osteogenic differentiation

“…All GSC culture media were changed every 3-4 days. The capacity for GSC osteogenic differentiation was examined on days 7, 14 and 21 [8].…”

“…Numerous previous studies relating to dentistry have analyzed the relationship between bacterial activity and inflammation resulting from periodontal disease which lead to problematic alveolar bone defects [5]. To overcome these defects, numerous techniques such as hydroxyapatite [6,7] or Carbonate-apatite bone graft [8] have been developed to stimulate and enhance the regeneration of alveolar bone.…”

“…Consequently, it was proposed to combine Gingival Stromal Progenitor Cells (GSCs) and Platelet Rich Fibrin (PRF) in order to stimulate and increase the regeneration of alveolar bone defect. By developing a combination of GSCs and PRF, it is possible to achieve the complete regeneration of alveolar bone defect [7,8]. GSCs osteogenic differentiation can be detected by Bone Sialoprotein-I (BSP-I) in its later stages [9][10][11].…”

Study of Human Amniotic Membrane Mesenchymal Stem Cells Using Gelatin and Alginate as Nontoxic Scaffolds

“…However, there are weaknesses in autogenous bone graft; for example, they are more expensive. Moreover, the required complex procedure depends on the location and size of the defect [3]. These donors are still unable to overcome the morphological and functional demands of three-dimensional structure, as well as the complicated construction of periodontal equipment, or the challenging features of peri-implant and alveolar bone defects [4].…”

“…Therefore, it is expected to obtain alternative methods of bone grafting that do not result in morbidity at the donor site. Donors should have unlimited availability and have the same bone regeneration capacity as autogenous bone grafts [3].…”