ABSTRACT
Abstract Objective: 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.
Subject(s)
Humans , Umbilical Cord , Lipopolysaccharides , Porphyromonas gingivalis , Cytotoxicity, Immunologic/immunology , Mesenchymal Stem Cells , Analysis of Variance , Flow Cytometry , Indonesia/epidemiologyABSTRACT
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.
Subject(s)
Animal Shells , Bone Substitutes , Brachyura , Durapatite , Fibroblasts , Animals , Bone Transplantation , Cell Survival , Cells, Cultured , Gingiva/cytology , Humans , In Vitro Techniques , Materials Testing , Microscopy, Electron, ScanningABSTRACT
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. 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.
Subject(s)
Animal Shells/chemistry , Brachyura , Durapatite/pharmacology , Fibroblasts/drug effects , Gingiva/cytology , Adult , Animals , Biocompatible Materials , Bone Transplantation , Humans , Materials Testing , Microscopy, Electron, ScanningABSTRACT
BACKGROUND: Studies of bone tissue engineering as a viable alternative to autogenous bone graft show promising results, although its mechanism and effectiveness remain only partially understood. PURPOSE: To explain the osteogenic differentiation of scaffold chitosan (Ch)-carbonate apatite (CA) in seeding with human amniotic mesenchymal stem cells (hAMSCs) on the regeneration of calvarial bone defects in rats. MATERIALS AND METHODS: Shitosan-Carbonate Apatite (Ch-CA) scaffold was created by means of a freeze-drying method. Twenty Wistar rats were randomly divided into two groups: control and treatment. Defects were created in the calvarial bone of each treatment group with a scaffold subsequently implanted. After 8 weeks, the rats were terminated for histology and immunohistochemistry examination. RESULTS: Expressions of vascular endothelial growth factor, bone morphogenetic protein2, Runt-related transcription factor 2 (RUNX2), and angiogenesis occurred earlier in the tissue-engineered group than that in the control group. An 8-week analysis also showed that the expression of RUNX2, alkaline phosphatase, osteocalcin, and collagen type 1 was at more elevated levels in the treatment group than that in the control group. CONCLUSION: These results showed that the combination of hAMSCs and Ch-CA scaffold may become one of the candidates for bone tissue engineering.
