METHODS: Twenty-four healthy young male rabbits were divided into two groups: T1, which underwent OTM and received normoxia-preconditioned GMSC, and T2, which underwent OTM and received hypoxia-preconditioned GMSC. A ligature wire was attached to the mandibular first molar and connected to a 50 g/mm2 closed coil spring, exerting force on the central incisor and left mandibular molar of the experimental animals. After 24 h of OTM, either normoxia- or hypoxia-preconditioned GMSC were injected into the gingiva of the samples in a single dose of 20 μl of phosphate-buffered saline (PBS). All samples were sacrificed on days 7, 14, and 28, and immunohistochemistry was performed to analyze the expression of RANK, RANKL, and OPG on the tension and compression sides.

METHODS: MON extract phytochemical analysis was conducted to examine active compounds such as flavonoids, saponins, quinones, alkaloids, tannins, terpenoids, and steroids. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay for antioxidant capacity was evaluated, and gas chromatography-mass spectrometry for the detection of volatile active compounds in MON extract was performed. Turax was used to create MON extract at concentrations of 1% and 2%, and then a particle size analysis was carried out. Prevotella intermedia (Pi), Porphyromonas gingivalis (Pg), Aggregatibacter actinomycetemcomitans (Aa), and Fusobacterium nucleatum (Fn) were tested for antibacterial activity of MON extract, comparing them with doxycycline as the reference drug and using the minimal inhibitory concentration (MIC), minimal bactericidal concentration (MBC), and diffusion zone methods.

MATERIALS AND METHODS:  Phytochemistry and liquid chromatography-high resolution mass spectrometry (LC-HRMS) were done to explore the active compounds in MLE. Chemistry screening and interaction, absorption, distribution, metabolism, and excretion (ADME), molecular docking simulation, and visualization of MLE active compounds as anti-inflammatory, antioxidant, and antibacterial were investigated in silico The inhibition zone of MLE against Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), and Fusobacterium nucleatum (Fn) as periodontopathogenic bacterias was performed by diffusion method. Doxycycline 100 mg was used as a positive control, as a treatment group, there were five groups, namely 0%, 25%, 50%, 75%, and 100% MLE.

RESULTS:  Alkaloid, saponin, flavonoid, triterpenoid, steroid, tannin, and quinone were detected in MLE. A high concentration of (-)epicatechin and coumaric acid (CA) were found in MLE. MLE in 100% concentration has the most effective ability to inhibit Fn, Pg, Aa growth in vitro. (-)-Epicatechin has a higher negative binding affinity than CA that can enhance heat shock protein (HSP)-30, HSP-70, HSP-90, interleukin-10, and FOXP3 and also inhibit interleukin-6, peptidoglycan, flagellin, and dectin in silico.

METHODS: A phytochemical analysis of 1 % RNE was carried out to examine the active substances possessed in it, such as flavonoids, quinone, saponin, alkaloids, tannins, terpenoids, and steroids. Nanoemulsion characterization was carried out using a particle size analyzer (PSA). The antibacterial activity of 1 % RNE toward Prevotella intermedia (Pi), Porphyromonas gingivalis (Pg), Aggregatibacter actinomycetemcomitans (Aa), and Fusobacterium nucleatum (Fn) was carried out to determine the minimum inhibitory concentration, minimum bactericidal concentration, and inhibitory zone compared with doxycycline as a positive control.

MATERIALS: AND METHODS:  Absorption, distribution, metabolism, excretion and toxicity prediction, molecular docking simulation, and visualization of chlorogenic acid (CGA) and coumaric acid (CA) as anti-inflammatory, antioxidant, and antibacterial were investigated in silico. Inhibition zone by diffusion method, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) of RGCBE extract against Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Fusobacterium nucleatum (Fn), and Prevotella intermedia (Pi) were done.

STATISTICAL ANALYSIS: the analysis of variance (ANOVA) difference test, and the post-hoc Tukey's Honest Significant Different (HSD) with a different significance value of p<0.05 RESULTS:  GCA and CA compounds are good drug molecules and it has low toxicity. Chlorogenic acid have higher binding activity than coumaric acid to tumor necrosis factor (TNF)-α, nuclear factor (NF)-κB, receptor activation NF-κB (RANK) and its ligand (RANKL), interleukin (IL)-6, IL-10, runt related transcription factor (RUNX2), receptor activator nuclear Kappa beta Ligand-osteoprotegrin osteocalcin (RANKL-OPG), osteocalcin, nuclear factor associated T-cell 1 (NFATc1), tartate resistant acid phosphatase (TRAP), peptidoglycan, flagellin, dectin, Hsp70, and Hsp10 protein. RGCB ethanol extract has high antioxidant ability and it has MIC, MBC, and inhibit the growth of Aa, Pg, Fn, and Pi at 50% concentration with significantly different (p=0.0001 and<0.05).

MATERIALS AND METHODS:  Forty-eight healthy, young male rabbits were divided into four groups: [-] OMF; [+] OMF; OMF with GMSCs normoxic-preconditioned; and OMF and GMSCs hypoxic-preconditioned. The central incisor and left mandibular molar in the experimental animals were moved, the mandibular first molar was moved mesially using nickel titanium (NiTi) and stainless steel ligature wire connected to a 50 g/mm2 light force closed coil spring. Allogeneic application of normoxic or hypoxic-preconditioned GMSCs was used in as many as 106 cells in a 20 µL phosphate buffered saline single dose and injected into experimental animals' gingiva after 1 day of OTM. On days 7, 14, and 28, all experimental animals were euthanized. Osterix, ALP, and osteopontin expressions were examined by immunohistochemistry.

MATERIALS AND METHODS: Twenty-four healthy male Wistar rats (R. norvegicus) were randomly divided into four groups: K1: blank, without treatment; K2: rats with an activated MEMF and PBS; P1: rats with an activated MEMF and HUCMSC-S; and P2: rats with an inactivated MEMF and HUCMSC-S. MEMF experimental animals was performed by applying a force of 0.42 N using a loop and observed for 28 days. The HUCMSC-S single dose was 2 x 106 cells (equivalent to 5 x 105 cells in 50 μl of aMEM) every two days application. After 28 days, the maxillary containing the midpalatal suture were harvested. Immunohistochemical analysis was performed to quantify the positive expression of BMP-2, BMP-4, BMP-7, VEGF, TGF-β, IGF-1 and FGF-2.

RESULTS: HUCMSC-S local application during active expansion (Group P1) significantly upregulated the expression of BMP-2, BMP-4, BMP-7, VEGF, TGF-β, IGF-1, and FGF-2 in the midpalatal suture on day 28 compared to the control groups (P ≤ 0.05).

MATERIALS AND METHODS: Twenty-eight healthy male Wistar rats, 1-2 months old and 250-300 g body weight, were divided into seven groups, namely K1: control group; K2: 100 μl LPS; K3: hyperglycemia (≥230 mg/dL); K4: 100 μl LPS with hyperglycemia; K5: LPS, hyperglycemia, and 100 μl NEP; K6: LPS, hyperglycemia, and 100 μl HUCMSCS; and K7: LPS, hyperglycemia, and 100 μl HUCMSCS and NEP. Escherichia coli`s LPS was used to induce osteolysis on the calvaria. NEP and HUCMSCS were formulated and collected, then injected subcutaneously on the calvaria. Hyperglycemia-induced streptozotocin 30 mg/kg injection for one week, intraperitoneally. All samples were sacrificed on day 8. Blood samples were collected to examine TRAP and Ctsk levels with enzyme-linked immunosorbent assay. NFATc1, sclerostin, and RANK-RANKL-OPG expression investigated with immunohistochemistry.

RESULTS: HUCMSCS and NEP administration inhibit TRAP and Ctsk levels, reduce NFATc1, sclerostin, and RANK-RANKL, but enhance OPG expression in LPS-induced osteolysis with hyperglycemia with significant differences between groups (P ≤ 0.05).