To show the cytotoxicity of Porphyromonas gingivalis lipopolysaccharide (LPS) on human umbilical cord mesenchymal stem cells (HUCMSCs) to better understand the characteristics for its application in regenerative procedures under periodontopathogen LPS influence. Material and Methods: Ultrapure Porphyromonas gingivalis LPS was used in this study. This research used a frozen stock HUCMSCs, previously confirmed by flow cytometry. The biological characteristics, such as cell morphology, proliferation, and protein expression, were screened. To check the cytotoxicity, HUCMSCs were cultured and divided into two groups, the control group and LPS group with various concentrations from 25 to 0.39 µg/mL. MTT assay was done and the cells were observed and counted. The significance level was set at 5%. Results: The percentage of living HUCMSCs on LPS group were not significantly different among concentrations (p>0.05) from 25 to 0.39 µg/mL, even though there were slight mean decrease between groups, but they were not significant. The duration of 24 hours of exposure of LPS does not significantly lower HUCMSCs viability. Conclusion: LPS does not affect the viability of HUCMSCs. The lower the concentration of LPS, the higher the viability of HUCMSCs.

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).

Background: Bone resorption due to tooth extraction leads to unpredictable bone volume for future prosthetics. Crab shells were promoted as a solution to prevent bone resorption, along with an effort to reduce biological waste. Purpose: This study aimed to analyze the expression of bone sialoprotein (BSP) and dentine matrix protein-1 (DMP1) in the wound healing process in tooth-extraction sockets after applying a crab shell-derived hydroxyapatite scaffold. Methods: The subjects (28 Cavia cobaya) were divided into control and treatment groups. The control group was left untreated, while the treatment group received a hydroxyapatite scaffold of Portunus pelagicus shell in the tooth socket. The expression of BSP and DMP1 was determined by immunohistochemical staining on days 7 and 14. One-way analysis of variance and Tukey’s honest significance difference test were used to find the groups with the most significant difference. Results: The highest mean expression of BSP and DMP1 was in the day 14 treatment group, while the lowest was in the day 7 control group. Conclusion: Administering hydroxyapatite scaffold derived from the Portunus pelagicus shell to the post-extraction sockets increased the expression of both BSP and DMP1.

Introduction:Crab shell (Portunus pelagicus) has the potential to be a source of hydroxyapatite biomaterials that used as bone grafts. Before clinical application, crab shell graft should be tested for its biocompatibility in vitro on human gingival fibroblast.Aim:This study aimed to determine the biocompatibility of Portunus pelagicus hydroxyapatite graft on human gingival fibroblast cell culture.Material and Methods:Human gingival fibroblast cell cultures were divided into control group and treatment group with the addition of hydroxyapatite graft powder from Portunus pelagicus at a concentration of 100 ppm, 50 ppm, and 25 ppm. The synthesis process of hydroxyapatite was conducted by heating at 1000°C then characterizing the compound with SEM-EDX. All samples were incubated in α-MEM medium, then were given MTT material. The cultures on the plate were examined using ELISA reader. The results were analyzed using a Oneway Anova.Results:The percentage of living cells throughout all treatment group shown results that exceeded the LD50 parameter. The highest percentage of living cells was at 25 ppm concentration group.Conclusion:The hydroxyapatite graft powder from crab shells is biocompatible with human gingival fibroblast cell culture.

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