Post Oral Administration of Epigallocatechin Gallate from Camelia sinensis Extract Enhances Vascular Endothelial Growth Factor and Fibroblast Growth Factor Expression during Orthodontic Tooth Movement in Wistar Rats.

Background:East Java green tea leaf (Camelia sinensis)possesed active compound such as EpigallocatechinGallate (EGCG) is well known for enhancing the boneremodelling through enhancement of VascularEndothelial Growth Factor (VEGF) and FibroblastGrowth Factors (FGF-2). Remodelling of alveolar boneis very important to obtain optimal Orthodontic ToothMovement (OTM) to align the tooth. Aim: Toinvestigate the expression of VEGF and FGF-2expression during OTM in Wistar rat afteradministration of EGCG from C. sinensis Extract(EGCG-CSE) Wistar rats. Material and Methods: Thisstudy was true experimental study with post-test onlycontrol group design. Twenty eight Wistar rats wererandomly selected and divided into four groupsaccordingly; K- group which did not get both EGCGCSE administration and OTM; K+ group with OTM for14 days, but no EGCG-CSE administration; 1 (T1) with4 days of OTM and 7 days of EGCG-CSEadministration; treatment group 2 (T2) with both 14days OTM and EGCG-CSE administration. Ten g2 force/mm of NiTi close coil spring was installedbetween the upper left molars and cental insicive tomove the molar mesially that induce OTM. All OTMthanimal model were terminated in the 14 days.Maxillary was isolated for immunohistochemistryinvestigation. Tukey Honest Significant Difference(HSD) was done after Analysis of Variance (ANOVA)test to investigate the significant difference betweengroups (p<0.05). Results: The highest positive VEGFexpression was found in the T2 in both area.Meanwhile, the highest positive FGF-2 expression wasfound in the K-group in both area. There weresignificant different of VEGF and FGF-2 expression inboth area between groups except T1 and T2.Conclusion: Post administration of EGCG-CSE canstimulate the VEGF and FGF-2 expression during OTMin Wistar rats.

Lipopolysaccharides' Cytotoxicity on Human Umbilical Cord Mesenchymal Stem Cells

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.

Cytotoxicity of Calcium Hydroxide on Human Umbilical Cord Mesenchymal Stem Cells

Abstract Objective: 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.

High Mobility Group Box 1 and Heat Shock Protein-70 Expression Post (-)-Epigallocatechin-3-Gallate in East Java Green Tea Methanolic Extract Administration During Orthodontic Tooth Movement in Wistar Rats

Abstract Objective: To investigate the expression of High Mobility Group Box 1 (HMGB1) and Heat Shock Protein-70 (HSP-70) during orthodontic tooth movement (OTM) after (-)- Epigallocatechin-3-Gallate (EGCG) in East Java Green Tea (Camelia Sinensis) Methanolic Extract (GTME) administration in vivo. Material and Methods: 28 Wistar rats (Rattus Novergicus) was used and divided into 4 groups accordingly: K- without EGCG and OTM; K+ with OTM, without EGCG for 14 days; T1with OTM for 14 days and EGCG for 7 days; treatment group 2 (T2) with OTM and EGCG for 14 days. OTM animal model was achieved through the installation of the OTM device by means of NiTi close coil spring with 10g force placed between the first incisor and first maxillary molars. The samples were terminated on Day 14. The pre-maxillary was isolated for the immunohistochemical examination. Analysis of Variance (ANOVA) then continued with Tukey Honest Significant Difference (HSD) (p<0.05) was performed to analyze the data. Results: The highest HMGB1 and HSP-70 expression were found in the K+ group pressure side, meanwhile the lowest HMGB1 and HSP-70 expression were found in K- group tension side in the alveolar bone. There was a significant decrease of HMGB1 and HSP-70 expression in T2 compared to T1 and K+ with significant between groups (p<0.05; p=0.0001). Conclusion: The decreased expression of HMGB1 and HSP-70 in alveolar bone of OTM wistar rats due to post administration of GTME that consisted EGCG.

Effect of Vitamin D during Orthodontic Tooth Movement on Receptor Activator of Nuclear Factor Kappa-Β Ligand Expression and Osteoclast Number in Pregnant Wistar Rat (Rattus novergicus)

Background: Female patients have the possibility tobecome pregnant during orthodontic treatment. VitaminD usually consumed by pregnant women. Estrogen andVitamin D could affect bone metabolism. Aim andObjectives: The aim of this study was to analyze theeffect of vitamin D during orthodontic movement inpregnant rats by Receptor Activator of Nuclear FactorKappa-Β Ligand (RANKL) expression and osteoclastnumber. Material and Methods: The experimentalobservational analytic study with post-test only controlgroup design and simple random sampling method wasconducted. 24-healthy-female Wistar rats were dividedinto 4 groups; K1: pregnant rats with orthodontic toothmovement and vitamin D on Day 7; K2: pregnant ratswith orthodontic tooth movement and vitamin D on Day14; K3: pregnant rats with orthodontic tooth movementwithout vitamin D on Day 7 and; K4: pregnant rats withorthodontic tooth movement without vitamin D on Day214. Nickle-Titanium coil spring with 10 g/mm forcewas placed between the incisors and the maxillarymolars. The RANKL expression and osteoclastsnumber were analyzed using Analysis of Variance(ANOVA) (p<0.05). Results: The highest osteoclastsnumber (8.494 ± 1.194), and RANKLexpression (7.967± 2.185) found in K1 group with significant betweengroups (p<0.05).Conclusions: Vitamin D increaseosteoclast number and RANKL expression duringorthodontic tooth movement in pregnant rats.

Regeneration of Salivary Gland Defects of Diabetic Wistar Rats Post Human Dental Pulp Stem Cells Intraglandular Transplantation on Acinar Cell Vacuolization and Interleukin-10 Serum Level

Abstract Objective: To investigate the regeneration of rat's salivary gland diabetic defect after intraglandular transplantation of Human Dental Pulp Stem Cells (HDPSCs) on acinar cell vacuolization and Interleukin-10 (IL-10). Material and Methods: HDPSCs isolated from the dental pulp of first premolars #34. HDPSCs from the 3rd passage was characterized by immunocytochemistry of CD73, CD90, CD105 and CD45. Twenty-four male Wistar rats, 3-month-old, 250-300 grams induced with Streptozotocin 30 mg/kg body weight to create diabetes mellitus (DM) divided into 4 groups (n=6); positive control group on Day-7; positive control group on Day-14; treatment group Day-7 (DM+5.105HDPSCs); treatment group on Day-14. On Day-7 and Day-14, rats were sacrificed. Histopathological examination performed to analyze acinar cells vacuolization while Enzyme-linked Immunoabsorbent Assay to measure IL-10 serum level. Data obtained were analyzed statistically using multiple comparisons Bonferroni test, Kruskal Wallis, Shapiro-Wilk and Levene's test result Results: The highest acinar cell vacuolization found in control group Day 14 (0.239 ± 0.132), meanwhile the lowest acinar cell vacuolization found in treatment group Day 7 (0.019 ± 0.035) with significant difference (p=0.003). The highest IL-10 serum level found in treatment group Day 14 (175.583 ± 120.075) with significant difference (p=0.001) Conclusion: Transplantation of HDPSC was able to regenerate submandibular salivary gland defects in diabetic rats by decreasing acinar cell vacuolization and slightly increase IL-10 serum level.

Correlation of Low CD4+ Counts with High Dental Caries Prevalence in Children Living with Perinatal HIV/AIDS Undergoing Antiretroviral Therapy

Abstract Objective: To investigate the correlation Cluster of Differentiation 4+ (CD4+) counts with a high prevalence of dental caries in Children Living with Perinatal Human Immunodeficiency Virus/Acquired Immunodeficiency Syndrome (CLWPHA). Material and Methods: An analytical observational research with a cross-sectional design was conducted at Tertiary General Hospital, Surabaya, Indonesia. Randomized total sampling consisted of 29 CLWPHA 1-12 years old at outpatient ward Infectious Disease Intermediate Unit (UPIPI) Tertiary General Hospital Doctor Soetomo Surabaya. Demographic details and recent CD4+ counts obtained from medical records. Decayed, missing, filled teeth (dmft) in primary teeth and DMFT in permanent teeth index were used. Pearson's and Spearman's Correlation test (p<0.05) were performed Results: Dental caries prevalence was 86.2%; meanwhile, the dmft/DMFT index score found were high (8.2 / 6.3). CD4+ count and caries showed significant correlation (r=-0.394, p<0.05). Tooth brushing frequency with caries showed a significant correlation (r=-0.419, p<0.05). Antiretroviral (ARV) therapy and caries showed an insignificant negative correlation (p<0.05) Conclusion: The high prevalence of dental caries in CLWPHA correlated with low CD4+ counts.