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: Prominent residual ridge is necessary to gain retention and stabilility for succesful prosthodontic treatment such as removable, fixed or implant. Spirulina is a natural substance that can help tissue healing and chitosan also a natural substance that reported to have the ability to help bone remodelling. The combination gel of spirulina and chitosan could be considered as an alternative material to maintain residual ridge height after tooth extraction. Purpose: The aim of study was to examine the effect of combination gel of Spirulina and chitosan on healing process of Cavia cobaya post tooth extraction socket by counting the amount of osteoclast, osteoblast and colagen as an indicator. Methods: Twenty eight cavia cobaya were divided into 4 groups. Insisive mandible extraction was done and the sockets were filled with 3% CMCNa for control groups, 3% spirulina chitosan 200 mg for group 1, 6% spirulina chitosan 200 mg for group 2, 12% spirulina chitosan 200 mg for group 3. After 30 days, histopathology examination was done by using microscope to count the amount of osteoclast, osteoblast and collagen. results: Data was analyzed by using Anova and Tukey HSD. For osteoclast, there was no significant different between every groups, while for osteoblast and collagen there was significant different between groups. The results showed that induction of combination gel spirulina chitosan was able to accumulate collagen fiber and resulting faster wound healing. Conclusion: Combination 12% gel spirulina chitosan 200 mg could be used as an alternative material for better bone remodeling after tooth extraction.

To examine the cytotoxicity of calcium hydroxide on human umbilical cord mesenchymal stem cells (HUCMSC) to understand the characteristics for use in regenerative dentistry procedures especially regenerative endodontics. Material and Methods: HUCMSC was isolated, cultured, and confirmed by flow cytometry. The biological characteristics, such as cell morphology, proliferation, and protein expression, were screened. To check the cytotoxicity, HUCMSC was cultured and divided into two groups, the control group (cultured in minimum essential medium (MEM) alpha) and calcium hydroxide group (cultured in MEM alpha and calcium hydroxide). Methyl-thiazole-tetrazolium (MTT) assay was done on different concentrations of calcium hydroxide (0.39 to 25 µg/mL) and the cells were observed and counted. One-way ANOVA test was used with a significance level set at 5%. Results: Flow cytometric analysis confirmed positive of CD73, CD90, CD105, negative of CD45 and CD34. A significant difference was found between the concentration of 6.25 and 3.125 µg/mL (p=0.004). There was no significant difference among 6.25, 12.5 and 25 µg/mL concentrations. There was also no significant difference among 0.39, 0.78, 1.56, and 3.125 µg/mL concentrations. Conclusion: Even though calcium hydroxide is a medicament of choice in clinical endodontics, it decreases the viability of HUCMSC. The lower the concentration of calcium hydroxide, the higher the viability of HUCMSC.

Background: Low levels of estrogen can cause osteoporosis and usually occur during a woman's menopausal phase. Osteoporosis can lead to bone resorption, the absence of osseointegration, and implant failure. The aim of this study is to determine the expression of transforming growth factor-beta 1 (TGF-β1), runt-related transcription factor (RUNX2), and osteoblasts in mandibular rats with low levels of estrogen. Materials and Methods: This study is an in vivo experimental research. Female Wistar rats ( n = 18) were divided into two groups: (1) Postsham surgery and (2) ovariectomy group. After 12 weeks, the rats were sacrificed to identify the level of estrogen, while histological analysis was conducted to determine the level of osteoblast and the expression of TGF-β1 and RUNX2. The data were analyzed using t -test ( P < 0.05). Results: There were significant lower levels of estrogen and osteoblast among the ovariectomy group compared to the postsham group ( P < 0.05). RUNX2 levels were found to be significantly higher in the ovariectomy group than that in the postsham group ( P < 0.05). However, there were no significant differences between TGF-β1 levels within the ovariectomy and postsham groups ( P > 0.05). Conclusion: Ovariectomy can lead to decreased osteoblastogenesis in mandibular bone by the reduced level of osteoblast and the increased expression of TGF-β1 and RUNX2.

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Objectives The aim of this study was to prove that human umbilical cord mesenchymal stem cell (hUCMSC) therapy conducted according to the mandibular osteoporotic model will increase Osterix (Osx) and bone morphogenetic protein-2 (BMP-2) expression, while reducing tartrate-resistant acid phosphatase (TRAP) expression. PKH26 labeling proves that mandibular bone regeneration is produced by hUCMSCs induction. Materials and Methods This study incorporated a true posttest only control group design. Twenty-five female Wistar rats were randomly divided into five groups consisting of the sham surgery (N) group, osteoporotic groups injected with gelatin for 4 weeks (G4) and 8 weeks (G8), and osteoporotic groups injected with hUCMSC-gelatin for 4weeks (SC4) and 8 weeks (SC8). All subjects were provided for BMP-2, Osx, and TRAP on immunohistochemistry examination and PKH-26 labeling. Statistical Analysis All data were analyzed using ANOVA and Tukey HSD tests with p < 0.05 being considered as statistically significant. Results Compared with other groups, the highest level of BMP-2 and Osx occurred in the sham surgery (N) and osteoporotic groups injected with hUCMSCs-gelatin (SC), while the lowest level of TRAP was found in SC4. During 4- and 8-week observation periods, the PKH 26 appeared green (fluorescent). Conclusions hUCMSC demonstrates high-osteogenic activity and increased osteoporotic mandibular bone regeneration, as shown by increased expression of Osx and BMP-2 and decreased TRAP expression. From the labeling, PKH-26 proved that viable hUCMSCs in gelatin solvent can be present in the mandibular bone and be capable of promoting osteogenic differentiation and increasing mineralization and bone formation in the osteoporotic mandibular bone.

Background: Diabetes Mellitus is a systemic disease characterized by an increase in blood glucose which, in the long term, enhances advanced glycation end product and leads to impaired osteogenesis. In prosthodontics, the osteogenic process plays an important role in successful treatment. Purpose: The purpose of this study is to determine the effect of Advanced Glycation End products (AGEs) present in Human Umbilical Cord Mesenchymal Stem Cells (hUCMSCs) on osteogenesis and calcification. Materials and Methods: MSCs isolated from human umbilical cord were cultured and underwent expansion up to passage 5. The research sample was divided into two sub-groups; a treatment group (osteogenic medium+AGE-BSA medium) and a control group (osteogenic medium) each of which underwent three replications. Samples were examined immunocytochemically on days 1, 3, 7, 8, 9, 12, 14 and 21 to quantify the level of RUNX2 expression. Alizarin red staining was performed on day 21. Results: In the treatment group, RUNX2 expression increased on day 3, reaching a peak on days 7 and 14. That expression decreased on day 8. In the control group, the expression of RUNX2 reached its peak on day 8 before decreasing on day 9. The presence of alizarin red indicated calcification in the control medium, but less mineralization in the treatment group. Conclusion: The research indicated that AGE-BSA enhances the production of RUNX2 expression in hUCMSCs at both the initial and maturation stages. This finding was supported by the results of alizarin red staining which indicated that increased levels of RUNX2 produced less mineralization.

Background:Prolongation of the inflammatory process in hyperglycemic interferes with bone formation, inhibits the healing process, and triggers bone resorption. A combination of spirulina and chitosan in the tooth socket of Rattus norvegicus is expected to promote the bone remodeling process. This study aimed to evaluate the effect of spirulina and chitosan on angiogenesis, osteoclast, and osteoblast cell in tooth socket models of type 1 diabetes.Materials and Methods:A laboratory-based experiment involving 36 R. norvegicus, divided into three groups (nondiabetes mellitus (DM), uncontrolled DM, and controlled DM) and further divided into six subgroups. The controlled groups (K1, K2, and K3) were induced with 3% carboxymethyl cellulose Na, while the treated groups were induced with 12% spirulina and 20% chitosan. On the 14th day, the mandibles of the rats were removed. The capillary lumen, osteoblasts, and osteoclast cells were counted by hypothalamic–pituitary–adrenal examination and the results analyzed by means of Shapiro–Wilk, Levene's, one-way ANOVA, and post hoc Tukey's honestly significant difference test.Results:There was a significant increment in the number of capillary lumen, osteoblast cells, and a decrease in osteoclasts in all three treated groups (P1, P2, and P3).Conclusions:A combination of spirulina and chitosan can effectively promote the healing process in postextraction sockets of type 1 DM R. norvegicus.