Hyperbaric Oxygen Therapy to Minimize Orthodontic Relapse in Rabbits.

OBJECTIVES: The purpose of the present study was to discover how hyperbaric oxygen therapy (HBOT) could reduce orthodontic relapse by altering the expressions of hypoxia-inducible factor (HIF)-1 messenger ribonucleic acid (mRNA), type I collagen (Col I), and matrix metalloproteinase-1 (MMP-1) in the gingival supracrestal fibers in rabbits. MATERIALS AND METHODS: This study involved 44 male rabbits (Oryctolagus cuniculus) randomly divided into the normal group (K0), the orthodontic group without HBOT (K1), and the orthodontic group with HBOT (K2). Following orthodontic separation of the two upper central incisors, a retention phase and relapse assessment were performed. The HBOT was performed for a period of 2, 4, 6, 8, and 10 days after retention. HIF-1α transcription was assessed employing real-time polymerase chain reaction, whereas Col I and MMP-1 proteins were examined using immunohistochemistry. The orthodontic relapse was measured clinically using a digital caliper. STATISTICAL ANALYSIS: We used the one-way analysis of variance followed by Tukey's post hoc for multiple comparisons to measure differences between pairs of means; a p-value of 0.05 was considered statistically significant. RESULTS: HBOT significantly increased the HIF-1α mRNA expression (p = 0.0140), increased Col I (p = 0.0043) and MMP-1 (p = 0.0068) on the tensioned and pressured side of the gingival supracrestal fibers, respectively, and clinically decreased the relapse (p = 3.75 × 10-40). CONCLUSION: HBOT minimizes orthodontic relapse by influencing HIF-1α expression, collagen synthesis (Col I), and degradation (MMP-1). This result suggests that HBOT has the potential to be used as an adjunctive method in the orthodontic retention phase.

RANK-RANKL-OPG expression after gingival mesenchymal stem cell hypoxia preconditioned application in an orthodontic tooth movement animal model.

Background: The expression of receptor activator of Nuclear Factor Kappa Beta (RANK) and its ligand (RANKL), as well as osteoprotegrin (OPG), in the alveolar bone (AB), may improve bone remodeling during orthodontic tooth movement (OTM). It is hypothesized that hypoxia-preconditioned gingival mesenchymal stem cells (GMSC) may be more effective than normoxia-preconditioned GMSC in this regard. This study aims to investigate the expression of RANK, RANKL, and OPG in the compression and tension sides of AB after allogeneic administration of GMSC that were normoxia or hypoxia-preconditioned in rabbits (Oryctolagus cuniculus) undergoing OTM. 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. Results: The expressions of RANK-RANKL-OPG in the alveolar bone of the compression and tension sides were significantly different during the 14-day period of OTM following allogeneic administration of GMSC that were normoxia or hypoxia-preconditioned (p < 0.05). Conclusion: The expression of RANK-RANKL was significantly increased on the compression side of the alveolar bone during OTM after the administration of hypoxia-preconditioned allogeneic GMSC but not on the tension side. Conversely, RANKL-OPG expression was enhanced on the tension side but not on the compression side, as observed through immunohistochemical analysis in vivo.

Gingiva Mesenchymal Stem Cells Normoxic or Hypoxic Preconditioned Application Under Orthodontic Mechanical Force on Osterix, Osteopontin, and ALP Expression.

OBJECTIVES: The aim of this article was to investigate Osterix, ALP, and osteopontin expression in the compression and tension sides of alveolar bone after the application of normoxic/hypoxic-preconditioned GMSCs in rabbits (Oryctolagus cuniculus) induced with OMF. 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. RESULTS: Osterix, ALP, and osteopontin expressions were significantly different after allogeneic application of hypoxic-preconditioned GMSCs than normoxic-preconditioned GMSCs in the tension and compression of the alveolar bone side during OMF (p < 0.05). CONCLUSION: Osterix, ALP, and osteopontin expressions were significantly more enhanced post-transplantation of GMSCs with hypoxic-preconditioning than after transplantation of normoxic-preconditioned GMSCs in rabbits (O. cuniculus) induced with OMF.

Gingival Mesenchymal Stem Cells Metabolite Decreasing TRAP, NFATc1, and Sclerostin Expression in LPS-Associated Inflammatory Osteolysis In Vivo.

OBJECTIVE: Bone is a dynamic tissue that undergoes remodeling. During bone remodeling, there are transcription factors such as nuclear factor-activated T cells-1 (NFATc1), sclerostin, and tartrate-resistant acid phosphatase (TRAP) that are released for bone resorption. Metabolite from gingival mesenchymal stem cells (GMSCs) has the ability to activate proliferation, migration, immunomodulation, and tissue regeneration of bone cells and tissues. Furthermore, the aim of this study is to investigate the metabolite of GMSCs' effect on expression of NFATc1, TRAP, and sclerostin in calvaria bone resorption of Wistar rats. MATERIALS AND METHODS: Twenty male healthy Wistar rats (Rattus norvegicus), 1 to 2 months old, 250 to 300 g body were divided into four groups, namely group 1 (G1): 100 µg phosphate-buffered saline day 1 to 7; group 2 (G2): 100 µg lipopolysaccharide (LPS) day 1 to 7; group 3 (G3): 100 µg LPS + 100 µg GMSCs metabolite day 1 to 7; and group 4 (G4): 100 µg GMSCs metabolite day 1 to 7. Escherichia coli LPS was used to induce inflammatory osteolysis on the calvaria with subcutaneous injection. GMSCs metabolite was collected after passage 4 to 5, then injected subcutaneously on the calvaria. All samples were sacrificed on the day 8 through cervical dislocation. The expression of TRAP, NFATc1, and sclerostin of osteoclast in the calvaria was observed with 1,000× magnification. STATISTICAL ANALYSIS: One-way analysis of variance and Tukey honest significant different were conducted to analyze differences between groups (p < 0.05). RESULTS: The administration of GMSCs metabolite can significantly decrease TRAP, NFATc1, and sclerostin expression (p < 0.05) in LPS-associated inflammatory osteolysis calvaria in Wistar rats (R. norvegicus). There were significantly different TRAP, NFATc1, and sclerostin expressions between groups (p < 0.05). CONCLUSION: GMSCs metabolite decrease TRAP, NFATc1, and sclerostin expression in LPS-associated osteolysis calvaria in Wistar rats (R. norvegicus) as documented immunohistochemically.

Oral Microbiome Profiles and Inflammation in Pregnant Women Who Used Orthodontic Appliances.

It is common for women to undergo orthodontic treatment during pregnancy, especially through the use of fixed orthodontic devices. In changing the oral microbiome profile, it is crucial to increase the immune responses of pregnant women using fixed orthodontics; however, changes in the microbiomes of pregnant women with orthodontic appliances can be adjusted. Therefore, we aimed to conduct research on the oral cavity microbiome profiles, specifically IL-6 and TNF-α, of pregnant women using fixed orthodontic appliances. We proposed an observational analysis of 30 third-trimester pregnant women. OHI-S was recorded, saliva collection was performed using the passive drool method for IL-6 and TNF-α, and analysis and mucosal swabs were used to determine the oral microbiome profile. Kruskal−Wallis and post hoc Bonferroni tests were used to identify any significant differences with values of p < 0.05. Of these pregnant women, those with orthodontic appliances developed 10 types of bacteria at similar levels (>80%) from the genera Streptococcus, Lactobacillus, and Veillonella. There was no difference between the oral microbiomes of the control group and the pregnant women with a history of orthodontic appliance use. While the level of TNF-α in the women with orthodontic appliances was higher compared with the control group who had never used orthodontic appliances (p < 0.05), there was no difference in the IL-6 levels. The IL-6 and microbiome profile produced normal results, so the use of orthodontic appliances during pregnancy should be allowed with conditions. Pregnant women with orthodontic appliances must keep the oral cavity clean and their appliances well-maintained to avoid oral problems.

The Short-Term Effect of Active Skeletonized Sutural Distractor Appliance on Temporomandibular Joint Morphology of Class III Malocclusion Subjects.

OBJECTIVES: The purpose of this study was to evaluate the short-term effect of active skeletonized sutural distractor (ASSD) appliance on temporomandibular joint morphology of class III malocclusion subjects. MATERIALS AND METHODS: This was a prospective interventional study. Cone-beam computerized tomography (CBCT) images of 22 patients were taken before and after treatment by using Planmeca Promax 3D CBCT machine version 2.9.2 (Planmeca OY Helsinki, Finland). The condylar width, height, length, roof of glenoid fossa thickness, and all joint spaces were measured. The condylar position was determined based on Pullinger and Hollander formula. The condylar shape was determined as per Kinzinger et al. The condylar volume was calculated by using Mimics software (Materialize, Belgium). STATISTICAL ANALYSIS: Data analysis was performed by using SPSS software version 24. Wilcoxon paired signed-rank test was used to compare the difference in temporomandibular joint morphology and condylar volume between pre- and post-treatment measurements. Chi-square test was used to compare the condylar position and shape. RESULTS: The superior (p = 0.000 on the right side, p = 0.005 on the left side) and posterior joint spaces (p = 0.000 on both sides) were decreased after the treatment, respectively. The condyles were rotated upward and backward, thereby increasing the anterior joint spaces (p = 0.000 on both sides) after the treatment. The condylar volume increases after treatment, but no significant differences were observed (p = 0.903 on the right side, p = 0.062 on the left side). CONCLUSION: The significant changes were observed in joint spaces. The condyles were more anteriorly placed before treatment. Condylar position and shape alter in response to ASSD treatment. The condylar volume did not show any significant change.

Effect of Caffeic Acid Phenethyl Ester Provision on Fibroblast Growth Factor-2, Matrix Metalloproteinase-9 Expression, Osteoclast and Osteoblast Numbers during Experimental Tooth Movement in Wistar Rats (Rattus norvegicus).

OBJECTIVES: To investigate the effect of caffeic acid phenethyl ester (CAPE) provision on matrix metalloproteinase-9 (MMP-9), fibroblast growth factor-2 (FGF-2) expression, osteoclast and osteoblast numbers during experimental orthodontic tooth movement (OTM) in male Wistar rats (Rattus norvegicus). MATERIALS AND METHODS: Forty-eight healthy male Wistar rats (R. norvegicus), 16 to 20 weeks old with 200 to 250 g body weight (bw) were divided into several groups as follows: K1: OTM for 3 days; K2: OTM for 7 days; K3: OTM for 14 days; KP1: OTM and CAPE for 3 days; KP2: OTM and CAPE for 7 days; and KP3: OTM and CAPE for 14 days. A nickel titanium closed coil spring 8.0 mm long with 10 g/mm2 was installed between the upper left first molar and upper central incisor to move molar mesially. CAPE provision with a dose of 20 mg/kg bw of animal studies was done per orally. Immunohistochemistry was done to examine MMP-9 expression and osteoclast number in compression side as well as FGF-2 expression and osteoblast number in tensile side of the OTM. STATISTICAL ANALYSIS: One-way analysis of variance test and Tukey's honest significant difference test were performed to determine the difference between the groups (p < 0.05). RESULTS: MMP-9 expression and osteoclast numbers in the compression side were significantly different between the groups. Similarly, FGF-2 expression and osteoclast numbers in the tensile side were significantly different between the groups. CONCLUSIONS: CAPE provision during OTM increases the number of osteoblasts and the FGF-2 expression significantly in the tensile side. Osteoclast numbers and MMP-9 expression significantly decrease in the compression side.

Gingival-Derived Mesenchymal Stem Cell from Rabbit (Oryctolagus cuniculus): Isolation, Culture, and Characterization.

OBJECTIVE: This study aims to confirm whether the GDMSCs isolated from rabbit's (Oryctolagus cuniculus) gingiva are mesenchymal stem cells (MSCs). MATERIALS AND METHODS: This study design was partly quasi-experimental with an observational design. GDMSCs were isolated from the gingiva of healthy male rabbits (O. cuniculus) (n = 2), 6 months old, and 3 to 5 kg of body weight. The specific cell surface markers of MSCs; clusters of differentiation (CD), namely, CD44, CD73, CD90, CD105, and CD200 expressions; and hematopoietic stem cell surface markers CD34 and CD45 were examined using flow cytometry and immunohistochemistry with immunofluorescence. The osteogenic differentiation of isolated GDMSCs was examined using alizarin red staining. RESULTS: GDMSCs in the fourth passage showed a spindle-like formation and fibroblast-like cells that attached to the base of the culture plate. GDMSCs were MSCs that positively expressed CD44, CD73, CD90, CD105, and CD200 but did not express CD34 and CD45 when examined using flow cytometry and immunohistochemical analysis. GDMSCs had osteogenic differentiation confirmed by calcified deposits in vitro with a red-violet and brownish color after alizarin red staining. CONCLUSION: GDMSCs isolated from the rabbits (O. cuniculus) were confirmed as MSCs in vitro documented using immunohistochemistry and flow cytometry. GDMSCs can differentiate into osteogenic lineage in vitro that may be suitable for regenerative dentistry.

Perspectives of Indonesian Orthodontists on the Ideal Orthodontic Treatment Time.

OBJECTIVE: The aim of this study was to explore the types of orthodontic treatment provided by Indonesian orthodontists and to analyse their perspectives on the ideal time to initiate orthodontic treatment. MATERIALS AND METHODS: A cross­sectional survey was conducted using the Google Drive questionnaire template. This electronic questionnaire was sent to a sample of orthodontists across different regions of Indonesia. The participants were asked to report the stage at which they would start orthodontic treatment, as well as answer questions about occlusal abnormalities and functional problems. Descriptive statistics for all variables were determined, including both practice characteristics and orthodontic treatment timing. RESULTS: A total of 152 orthodontists agreed to participate in the study, of which 64.5% were female and 35.5% were male. Indonesian orthodontists prefer two-phase orthodontic treatment. Sucking habits and open bite were found to be the most frequent indications for treatment in the primary dentition. Anterior crossbite was found to be the most frequent indication for treatment during the early mixed dentition stage. Severe Class II was found to be the most frequent indication for treatment during the late mixed dentition stage. Indonesian orthodontists are more concerned about impacted canines and midline diastema than other occlusal deviations in the permanent dentition. CONCLUSION: Based on the results of this study, we can conclude that Indonesian orthodontists favor two-phase orthodontic treatment. They also prefer to treat sucking habits and open bite in the primary dentition, anterior crossbite in the early mixed dentition, and severe Class II during the late mixed dentition stage.

The effect of low-level light therapy on orthodontic tooth movement rate, heat shock protein 70, and matrix metallopreteinase 8 expression: Animal study.

BACKGROUND: The aim of this study was to determine the effect of low-level light therapy (LLLT) on orthodontic tooth movement (OTM) rate, heat shock protein 70 (HSP-70) expression, and matrix metalloproteinase 8 (MMP-8) expression. MATERIALS AND METHODS: In this experimental study twenty-four male guinea pigs were randomly divided into three groups (n = 8): control group (K) without orthodontic force and LLLT; treatment group 1 (T1) with orthodontic force, and treatment group 2 (T2) with orthodontic force and LLLT. The labial surfaces of both maxillary central incisors in treatments groups were installed with single-wing bracket before being inserted with close coil spring to give 10 g/cm2 orthodontic force. For the T2 group, 4 J/cm2 of LLLT was administered in the mesial-distal and labial-palatal regions for 3 min every day. On day 14, the gap between teeth was measured and immunohistochemistry staining was done to determine HSP-70 and MMP-8 expression. Data were analyzed using (IBM, New York, (ANOVA), followed by Turkey's HSD test to determine the differences between groups. Nonnormal distributed data would be analyzed using Kruskal-Wallis test, followed by Mann-Whitney test with P < 0.05 being performed. RESULTS: The gap between teeth in the T2 group was greater compared to T1 group (P = 0.00). However, there was a significant decrease of HSP-70 and MMP-8 expression in T2 group compared to T1 group in the tensile and compressive sides. CONCLUSION: LLLT intervention during orthodontic treatment could accelerate OTM rate and decreased HSP-70 and MMP-8 expression both in tension and in compressive side. Thus, LLLT interventions can be used as adjuvant therapy to shorten orthodontic treatment duration.

Soluble Human Leukocyte Antigen Molecules Detected in Orofacial Cleft Patients: A Case-Control Study

Abstract Objective: To compare soluble HLA-C and HLA-DR molecules present in the plasma of orofacial cleft and non-orofacial cleft populations. Material and Methods: Orofacial cleft patients were recruited using an accidental sampling approach (n=15). Peripheral blood was collected from the participants and processed for Enzyme Linked Immunosorbent Assay (ELISA) against HLA-C and HLA-DR with specific antibodies. The absorbance was calculated utilizing ELISA reader. Data were statistically analyzed using an independent t-test to compare the disease and control groups. Results: The levels of soluble HLA-C and HLA-DR were significantly higher in the diseased group compared to the control group (p<0.05). Conclusion: The role of HLA molecules in non-communicable disease and congenital anomalies, particularly orofacial cleft, remains speculative despite the positive results of this study and those of previous investigations. It suggests that the variables examined may affect specific pathways involved in the pathogenesis of orofacial cleft, and predispose the individuals concerned to the oral cleft.

Receptor Activator of Nuclear Factor-Kappa Ligand and Osteoprotegerin Expressions on Hyperglycemic Wistar Rats (Rattus Novergicus) During Orthodontic Tooth Movement

Abstract Objective: To investigate the differences of receptor activator of nuclear factor-κB ligand (RANKL) and Osteoprotegerin (OPG) expressions between normoglycemic and hyperglycemic Wistar rats (Rattus Novergicus) during Orthodontic Tooth Movement (OTM). Material and Methods: This study was true experimental with post-test group only. Thirty-two healthy male Wistar rats, weighted around 200-250 grams, 12-20 weeks old, were used as OTM animal study. They were divided into 2 groups (n=16), normoglycemic rats (normal blood glucose 80-120 mg/dl) and hyperglycemic rats (>250 mg/dl) induced by Streptozotocin with a dose of 30 mg in PBS injection intraperitoneally. A NiTi closed coil spring was mounted between maxillary first molar and incisors with the light force 10gf/mm2 in both groups to induce OTM. The studied animals were then terminated on days 1, 3, 6, and 9, respectively, and premaxilla was extracted. RANKL and OPG expression were examined utilizing immunohistochemistry (IHC) analysis. One-way ANOVA and Tukey HSD (p<0.05) were utilized to analyze the differences in the expression of RANKL and OPG between groups. Results: The hyperglycemic group on day 1, 9 rats showed a significant increase in the expression of RANKL, whereas OPG expression decreased significantly on days 1, 3, and 9. Conclusion: There was a significant increase of RANKL expression and a decrease of OPG expression in hyperglycemic rats as documented immunohistochemically.

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.

Medicinal Signaling Cells Metabolite Oral Based as a Potential Biocompatible Biomaterial Accelerating Oral Ulcer Healing (In Vitro Study).

OBJECTIVE: Medicinal signaling cells metabolite (MSCM) is often considered medical waste even though it contains abundant growth factors, and advantageous micro- and macromolecules that can accelerate healing in oral ulcer.The purpose of this experimental laboratory study was to analyze the biocompatibility and potential of MSCM, (oral based) to accelerate healing in oral ulcer (in vitro). MATERIALS AND METHODS: MSCM (oral based) was obtained by mixing 10 mL of MSCM and 2% of carboxymethyl cellulose sodium. 3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (or MTT assay) was obtained using human gingival somatic cell culture to examine cell viability treated with MSCM (oral based). Fourier transform infrared spectroscopy was performed to know the functional structure and composition of MSCM (oral based). To know the elemental composition of MSCM (oral based), energy-dispersive X-ray analysis was performed. Scratch test was performed to know the ability of MSCM (oral based) to increase human somatic cell proliferation. RESULTS: MSCM (oral based) has good cell viability. MSCM (oral based) administration accelerated the proliferation of human somatic cell culture after 12-hours in vitro. MSCM (oral based) has carboxylic acids and derivatives chemical bond. MSCM (oral based) mostly contained carbon and potassium but did not contain heavy metal substances. CONCLUSIONS: MSCM (oral based) has a biocompatible and potential ability to accelerate healing in oral ulcer in vitro. It would be useful in daily clinical practice in treating traumatic oral ulcer.

A comparison of antibacterial inhibitory effect on Streptococcus mutans and tensile strength between chitosan-based bonding adhesives and commercial products.

BACKGROUND: Adhesive bonding is the material used to attach a bracket to the enamel surface of the tooth. Streptococcus mutans contributes to enamel demineralization during orthodontic treatment. OBJECTIVES: To analyze the antimicrobial inhibitory effect of Streptococcus mutans bacteria and tensile strength of chitosan and CaCO3-based adhesive bonding material. MATERIALS AND METHODS: The investigation constituted laboratory experimental research featuring analytical observation and a random sampling method. The antibacterial inhibitory effect of chitosan and CaCO3-based adhesive bonding against Streptococcus mutans involved six groups: two control groups using commercial light cure and self-cure adhesive bonding products and four groups using adhesive bonding consisting of 75% CaCO3 + 17.6% Bis-GMA + 22.4% MMA with various percentages of chitosan composition (A1: 25%, A2: 50%, A3: 75%, and A4: 100%) each group consisting of two samples (n = 12). A diametric test was conducted consisting of three samples (n = 15) to measure the tensile strength of each group. Data were analyzed by a combination of one-way analysis of variance and least significant difference tests. RESULT: The antibacterial inhibitory effect showed significant differences between groups (A1: 2.9467 ± 0.4163, A2: 3.6500 ± 0.6245, A3: 5.1267 ± 0.2517, A4: 4.7267 ± 0.9238; P = 0.0000; P < 0.05). A diametric tensile strength test confirmed significant differences between groups (A1: 7.2733 ± 5.0046, A2: 6.7667 ± 4.4346, A3: 6.4533 ± 2.9994, A4: 1.0058 ± 1.0058, K1: 15.6167 ± 3.1250; P = 0.009; P < 0.05). CONCLUSION: Chitosan-based adhesive bonding with good tensile strength has an antibacterial inhibitory effect against Streptococcus mutans.

The Densitometric Analysis of Protein Pattern in Cleft Lip and Palate Patients.

OBJECTIVES: Cleft lip and palate (CLP) belongs to the congenital anomaly that is clinically seen as cleft in lip, alveolar bone, palate, and nasal septum. The patients suffer from esthetic and various functional defects. CLP is resulted from impaired palatogenesis during the embryonic phase. The etiology of CLP is influenced by genetic, environmental, and combination of both. According to the literature, CLP is highly associated with defect in interferon regulatory factor 6 (IRF6) and poliovirus receptor-like (PVRL1) genes. The present study aimed to investigate the total protein profile and to identify protein IRF6 and PVRL1 in plasma of CLP patients. MATERIALS AND METHODS: Dot-Blot analysis was performed to identify protein target of IRF6 and PVRL1. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was performed in gel concentration 12% using plasma of CLP patients, their parents, and control population. The gels were stained by Coomassie blue afterward. Gels were analyzed through ImageLab 5.2.1 software. RESULTS: The intensity of major bands in CLP patients was darker than control group, but remains similar to the parents group. The target protein IRF6 and PVRL1 were positively identified through Dot-Blot. Retardation factor value was significantly different in major bands of CLP patients compared to control group. CONCLUSION: There pattern of protein profile in CLP patients was different compared to non-CLP.

The Role of Low-Intensity Biostimulation Laser Therapy in Transforming Growth Factor ß1, Bone Alkaline Phosphatase and Osteocalcin Expression during Orthodontic Tooth Movement in Cavia porcellus.

OBJECTIVE: The aim of this study is to analyze the low-intensity laser therapy (LILT) biostimulation mechanism as adjuvant therapy within orthodontic treatment as a means of accelerating bone remodeling by transforming growth factor ß1 (TGF-ß1), bone alkaline phosphatase (BALP), and osteocalcin (OSC) expression. MATERIALS AND METHODS: An analytical experimental method incorporating a posttest only randomized the control group design. The sample consisted of 24 3- to 4-month-old male Cavia porcellus weighing between 300 and 500 g divided into three groups (group 1: control, group 2: received orthodontic treatment, and group 3: received orthodontic treatment with irradiation LILT). LILT biostimulation at a dose of 4 joule/cm2 was performed daily for 3 min on the mesial-distal labial-palatal of the first dextra and sinistra incisor for 2 weeks. The TGF-ß1, BALP, and OSC expression was subjected to immunohistochemical analysis. An analysis of variance with multiple comparison, a Tukey's honestly significant difference test, a Kruskal-Wallis test, and a Wilcoxon-Mann-Whitney test were all performed (p < 0.05). RESULTS: TGF-ß1 expression was significantly different (p = 0.047; p < 0.05) in the tension area, but not in the compression side (p = 0.154; p > 0.05). BALP expression was significantly different in both the tension (p = 0.009) and compression areas (p = 0.005; p < 0.05). OSC expression was significantly different (p = 0.034; p < 0.05) in the tension side, but not in the compression area (p = 1.194; p > 0.05). CONCLUSION: LILT biostimulation can increase TGF-ß1, BALP, and OSC expression during orthodontic tooth movement.

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.

Osteogenic potential of gingival stromal progenitor cells cultured in platelet rich fibrin is predicted by core-binding factor subunit-α1/Sox9 expression ratio ( in vitro).

Background: Alveolar bone defect regeneration has long been problematic in the field of dentistry. Gingival stromal progenitor cells (GSPCs) offer a promising solution for alveolar bone regeneration. In order to optimally differentiate and proliferate progenitor cells, growth factors (GFs) are required. Platelet rich fibrin (PRF) has many GFs and can be easily manufactured. Core-binding factor subunit-α1 (CBF-α1) constitutes a well-known osteogenic differentiation transcription factor in SPCs. Sox9, as a chondrogenic transcription factor, interacts and inhibits CBF-α1, but its precise role in direct in vitro osteogenesis remains unknown. GSPCs cultured in vitro in PRF to optimally stimulate osteogenic differentiation has been largely overlooked. The aim of this study was to analyze GSPCs cultured in PRF osteogenic differentiation predicted by CBF-α1/Sox9. Methods: This study used a true experimental with post-test only control group design and random sampling. GPSCs isolated from the lower gingiva of four healthy, 250-gram, 1-month old, male Wistar rats ( Rattus Novergicus) were cultured for two weeks, passaged every 4-5 days. GSPCs in passage 3-5 were cultured in five M24 plates (N=108; n=6/group) for Day 7, Day 14, and Day 21 in three different mediums (control negative group: αModified Eagle Medium; control positive group: High Glucose-Dulbecco's Modified Eagle Medium (DMEM-HG) + osteogenic medium; Treatment group: DMEM-HG + osteogenic medium + PRF). CBF-α1 and Sox9 were examined with ICC monoclonal antibody. A one-way ANOVA continued with Tukey HSD test (p<0.05) based on Kolmogorov-Smirnov and Levene's tests (p>0.05) was performed. Results: The treatment group showed the highest CBF-α1/Sox9 ratio (16.00±3.000/14.33±2.517) on Day 7, while the lowest CBF-α1/Sox9 ratio (3.33±1.528/3.67±1.155) occurred in the control negative group on Day 21, with significant difference between the groups (p<0.05). Conclusion: GSPCs cultured in PRF had potential osteogenic differentiation ability predicted by the CBF-α1/sox9 ratio.

Bone alkaline phosphatase and osteocalcin expression of rat's Gingival mesenchymal stem cells cultured in platelet-rich fibrin for bone remodeling (in vitro study).

OBJECTIVE: The aim of this study was to analyze the osteogenic differentiation of rat GMSCs cultured in PRF for bone remodeling. MATERIALS AND METHODS: GMSCs were isolated from the lower gingival tissue of four healthy, 250 g, 1-month old, male rats (Rattus norvegicus) cut into small fragments, cultured for 2 weeks, and subsequently passaged every 4-5 days. GMSCs isolated in passage 3 were characterized by CD34, CD45, CD44, CD73, CD90, and CD105 using fluorescein isothiocyanate immunocytochemistry (ICC) examination. GMSCs in passage 3-5 cultured in five M24 plates (N = 108; n = 6/group) for 7, 14, and 21 days with three different mediums as follows: Control (-) group: α-Modified Eagle Medium; Control (+) group: High-dose glucose Dulbecco's Modified Eagle's Medium (DMEM-HG) + osteogenic medium; and treatment group: DMEM-HG + osteogenic medium + PRF. GMSCs were osteogenic differentiation cultured in vitro in three different mediums by bone alkaline phosphatase (BALP) and osteocalcin (OSC) marker using ICC monoclonal antibody. STATISTICAL ANALYSIS USED: The one-way analysis of variance was performed (P < 0.05) based on Shapiro-Wilk and Levene's tests (P > 0.05). RESULTS: GMSCs were shown to present + CD44, +CD73, +CD90, +CD105 and - CD34, - and CD45 expression as MSCs markers. The treatment group showed the highest BALP expression (16.00 ± 1.732) on day 7, while OSC expression (13.67 ± 2.309) on day 21 showed the statistically significant difference between groups (P < 0.05). CONCLUSION: GMSCs cultured in PRF demonstrated potential osteogenic differentiation ability capable of accelerating in vitro bone remodeling by enhancing BALP and OSC expression.