Determination of the compressive modulus of elasticity of periodontal ligament derived from human first premolars.

Purpose: There are two commonly cited modulus of elasticity of the human periodontal ligament (EPDL), i.e., 6.89 ✕ 10-5 GPa (E1) and 6.89 ✕ 10-2 GPa (E2), which are exactly 1000-fold different from each other. This study aims to clarify the ambiguity of the two EPDL used for simulations and determine a more accurate EPDL value of human first premolars using experimental and simulation approaches. Methods: Numerical simulations using finite element analysis were performed to analyze PDL deformation under an average Asian occlusal force. To confirm the results, simple and multi-component, true-scale 3D models of a human first premolar were used in the simulations. Finally, a compression test using a universal testing machine on PDL specimens was conducted to identify the compressive EPDL of human first premolars. Results: The simulation results from both models revealed that E1 was inaccurate, because it resulted in excessive PDL deformation under the average occlusal force, which should not occur during mastication. Although the E2 did not lead to excessive PDL deformation, it was obtained by an error in unit conversion with no scientific backing. In contrast, the compression test results indicated that the compressive EPDL was 9.64 ✕ 10-4 GPa (E3). In the simulation, E3 did not cause excessive PDL deformation. Conclusion: The simulation results demonstrated that both commonly cited EPDL values (E1 and E2) were incorrect. Based on the experimental and simulation results, the average compressive EPDL of 9.64 ✕ 10-4 GPa is proposed as a more accurate value for human first premolars. Clinical significance: The proposed more accurate EPDL would contribute to more precise and reliable FEA simulation results and provide a better understanding of the stress distribution and deformation of dental materials, which will be beneficial to precision dentistry, orthodontics and restoration designs.

PTEN regulates proliferation and osteogenesis of dental pulp cells and adipogenesis of human adipose-derived stem cells.

OBJECTIVE: To investigate the role of phosphatase and tensin homolog (PTEN) in dental pulp cells (hDPs) and adipose-derived mesenchymal stem cells (hADSCs). MATERIALS AND METHODS: Genetic variant was identified with exome sequencing. The hDPs isolated from a patient with Cowden syndrome were investigated for their proliferation, osteogenesis, adipogenesis, and gene expression compared with controls. The normal hDPs and hADSCs were treated with the PTEN inhibitor, VO-OHpic trihydrate (VOT), to investigate the effect of PTEN inhibition. RESULTS: A heterozygous nonsense PTEN variant, c.289C>T (p.Gln97*), was identified in the Cowden patient's blood and intraoral lipomas. The mutated hDPs showed significantly decreased proliferation, but significantly upregulated RUNX2 and OSX expression and mineralization, indicating enhanced osteogenic ability in mutated cells. The normal hDPs treated with VOT showed the decreases in proliferation, colony formation, osteogenic marker genes, alkaline phosphatase activity, and mineral deposition, suggesting that PTEN inhibition diminishes proliferation and osteogenic potential of hDPs. Regarding adipogenesis, the VOT-treated hADSCs showed a reduced number of cells containing lipid droplets, suggesting that PTEN inhibition might compromise adipogenic ability of hADSCs. CONCLUSIONS: PTEN regulates proliferation, enhances osteogenesis of hDPs, and induces adipogenesis of hADSCs. The gain-of-function PTEN variant, p.Gln97*, enhances osteogenic ability of PTEN in hDPs.

Utilization of rapid antigen tests for screening SARS-CoV-2 prior to dental treatment.

Potential aerosols containing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viral particles can be generated during dental treatment. Hence, patient triage is essential to prevent the spread of SARS-CoV-2 in dental clinical settings. The present study described the use of rapid antigen tests for SARS-CoV-2 screening prior to dental treatment in an academic dental clinical setting in Thailand during the pandemic. The opinions of dental personnel toward the use of rapid antigen test screening prior to dental treatment were also assessed. From August 25 to October 3, 2021, dental patients who were expected to receive aerosols generating dental procedures were requested to screen for SARS-CoV-2 using a rapid antigen test before their treatment. A total of 7,618 cases completed the screening process. The average was 212 cases per day. Only five patients (0.07%) were positive for SARS-CoV-2 in the rapid antigen screening tests. All positive cases exhibited mild symptoms. For the questionnaire study, experienced dental personnel frequently and consistently agreed with the use of the rapid antigen test for SARS-CoV-2 screening, which made them feel safer during their patient treatment. However, implementing rapid antigen tests for SARS-CoV-2 may increase the total time spent on a dental appointment. In conclusion, a rapid antigen test could detect the infected individual prior to dental treatment. However, the specificity of rapid antigen tests for SARS-CoV-2 must be taken into account for consideration as a screening process before dental treatment. The enhanced infection control protocols in dental treatment must be consistently implemented.

The Prostacyclin Analog Iloprost Promotes Cementum Formation and Collagen Reattachment of Replanted Molars and Up-regulates Mineralization by Human Periodontal Ligament Cells.

INTRODUCTION: This study evaluated the use of the prostacyclin analog iloprost as a root surface treatment agent in promoting acellular cementum (AC) formation and collagen reattachment after tooth replantation in vivo. In addition, its effect on human periodontal ligament cell (hPDLC) mineralization was assessed in vitro. METHODS: First molars of 8-week-old Wistar rats were extracted. In 1 group, the root surfaces were treated with Hank's balanced salt solution (HBSS), and the other group's root surfaces were treated with 10-6 mol/L iloprost before replantation. At day 30, maxillae were prepared for micro-computed tomographic imaging and histomorphometric analysis. The effect of iloprost on mineralization by hPDLCs was analyzed by mineralized nodule formation and quantitative polymerase chain reaction at 7 and 14 days. RESULTS: Micro-computed tomographic imaging demonstrated a significant higher bone volume in the iloprost groups, whereas the HBSS groups had extensive bone and root resorption. Histologic analysis revealed deposition of a thick AC layer along the root in the iloprost group with well-organized periodontal ligament fibers inserted into the cementum. The HBSS group demonstrated more osteoclasts than the iloprost group. In vitro, iloprost-treated hPDLCs had a significantly increased RUNX2, OSX, BSP, and ALP gene expression that coincided with an increased deposition of mineralized nodules. These effects were abrogated by a PGI2 receptor inhibitor. CONCLUSIONS: Our results revealed that iloprost promoted PDL regeneration in replanted molars. Furthermore, resorption of the roots was decreased, whereas AC deposition was stimulated. Iloprost-treatment increased hPDLC mineralization and was mediated by PGI2 receptor activation. These observations indicate that iloprost may be a promising root surface treatment agent.

The impact of deproteinized bovine bone particle size on histological and clinical bone healing outcomes in the augmented sinus: A randomized controlled clinical trial.

OBJECTIVE: The effect of different deproteinized bovine bone mineral (DBBM) particle sizes on bone healing in maxillary sinus floor augmentation remains unclear. This study compared the newly formed tissue and angiogenesis-related bone healing after sinus floor augmentation using large or small DBBM particles. MATERIALS AND METHODS: Overall 32 patients were randomly divided into two groups using either large (1-2 mm) or small (0.25-1 mm) DBBM particles for sinus floor augmentation. After 6 months, the mineralized tissue volume was calculated using micro-computed tomography (micro-CT) analysis. The newly formed tissue composition was histomorphometrically analyzed. Angiogenesis was also examined by means of vascular endothelial growth factor (VEGF) expression. Implant failure and marginal bone loss were measured at a 1-year follow-up. Statistical analysis was performed using independent samples t-test. RESULTS: Micro-CT analysis demonstrated that grafting with large particles resulted in higher bone volume (6.99 ± 2.72 mm3 , p = 0.002) and Bone Volume/Tissue Volume (0.25 ± 0.1, p = 0.03) compared with small particles (3.76 ± 1.83 mm3 and 0.14 ± 0.13, respectively). Small particles showed higher non-mineralized tissue volume (26.31 mm3 ) compared with large particle group (17.4 ± 5.34 mm3 ) with p = 0.001. The histological data revealed significantly higher area of newly formed bone (32.15% ± 14.04% for the large particle and 15.99% ± 14.12% for the small particle groups, p = 0.004). Likewise, non-mineralized tissue was significantly greater in the small particle group (66.48% ± 20.97%) compared with the large particle group (44.36%, p = 0.016). Moreover, use of large particles resulted in a significantly higher VEGF staining intensity score and VEFG positive cells. No implant failure was recorded in both groups, while no difference was found in terms of marginal bone loss at the 1-year follow-up. CONCLUSIONS: Sinus floor augmentation using large DBBM particles resulted in more angiogenesis expression, higher bone volume, and new bone formation at 6 months after sinus augmentation. However, clinical outcomes with regards to implant placement were similar in both groups.

Taking 200 mg Vitamin C Three Times per Day Improved Extraction Socket Wound Healing Parameters: A Randomized Clinical Trial.

Vitamin C is essential for wound healing. However, there are no reports concerning the effect of a different dose of vitamin C on extraction wound size clinically. Therefore, the aim of this study was to investigate the effect of different oral vitamin C doses on extraction wound healing. A split-mouth, double-blind randomized clinical trial was performed in 42 patients who underwent symmetric bilateral noninfected premolar extraction. The patients were randomly divided into 3 groups, namely, P/600, P/1,500, and 600/1,500 (14 patients for each group); P/600: placebo vs. 600 mg vitamin C/d, P/1,500: placebo vs. 1,500 mg vitamin C/d, and 600/1,500: 600 mg vitamin C/d vs. 1,500 mg vitamin C/d. Patients were prescribed placebo or/and vitamin C three times a day for 10 days after each tooth extraction. Extraction wound size and pain score were evaluated. The wound assessment was performed on day 0, 7, and 21; and then the tooth on the other side was extracted using the same protocol. Pain score was recorded on the first three days after extraction. The reduced size of mesiodistal extraction wound in percentage reduction between day 0 and 7 of teeth receiving vitamin C 600 mg/d was more than that in placebo (P < 0.05). Pain scores on day 1-3 of teeth receiving vitamin C 600 mg/d were significantly lower than the placebo side (P < 0.05). Taking oral vitamin C 600 mg/d over three doses for 10 days after tooth extraction enhances extraction wound healing by reducing mesiodistal extraction wound and reduces postoperative pain.

A split-mouth randomized clinical trial to evaluate the effect of local and systemic administration of vitamin C on extraction wound healing.

A split-mouth single-blind randomized-controlled clinical trial study was designed to investigate the effect of local and systemic vitamin C administration on extraction wound healing. Thirty patients who underwent bilateral premolar extraction were randomly divided into three group pairs; group 1: control and systemic administration (Con/CSA), group 2: control and a combination of local and systemic administration (Con/CLSA), and group 3: systemic and a combination of local and systemic administration (CSA/CLSA). The vitamin C (600 mg) was taken by swallowing (systemic administration) or slow oral dissolution (combined local and systemic administration). The socket size and radiographic density were evaluated immediately after extraction, and 7 days and 21 days later. The results demonstrated that the percentage radiographic density of new bone formation in the socket did not differ significantly within each group. However, in the CSA and CLSA group there was an improvement of soft tissue healing based in terms of socket depth reduction at 21 days after extraction compared with the control (P < 0.05).

Rinsing with L-Ascorbic Acid Exhibits Concentration-Dependent Effects on Human Gingival Fibroblast In Vitro Wound Healing Behavior.

Vitamin C or L-ascorbic acid has diverse functions in the body, especially healing promotion in tissue injury via participating in the hydroxylation reactions required for collagen formation. Systemic administration of vitamin C plays an important role on gingival fibroblast proliferation and functions. Whether local or rinsing administration of vitamin C alters gingival fibroblast wound healing behavior remains unclear. The aim of this study was to investigate the rinsing effect of vitamin C on gingival fibroblast behavior utilizing an in vitro wound healing model. Primary human gingival fibroblasts isolated from gingival tissue were rinsed with medium containing various concentrations of vitamin C. The rinsing effect of vitamin C on in vitro wound healing was assessed using a scratch test assay. Cell migration, cell viability, and extracellular matrix gene expression were analyzed by transwell migration assay, MTT assay, and real-time RT-PCR, respectively. We found that rinsing with 10 or 20 µg/ml vitamin C significantly increased fibroblast migration (p ≤ 0.05). However, no significant effect was found in the cell viability or in vitro wound healing assays. In contrast, rinsing with 50 µg/ml vitamin C significantly delayed wound closure (p ≤ 0.05). Real-time PCR demonstrated that 50 µg/ml vitamin C significantly increased fibroblast expression of COL1, FN, IL-6, and bFGF. The data demonstrate that rinsing with vitamin C (10/20 µg/ml) accelerates fibroblast migration. However, 50 µg/ml of vitamin C increases the expression of COL1, FN, IL-6, and bFGF, which are related to fibroblast wound healing activity. Prescribing vitamin C with the appropriate duration and drug administration method should be determined to maximize its benefit.

Dentofacial Deformity Caused by Bulky Osteochondroma: Report of an Unusual Case and the Importance of Cone Beam Computed Tomography.

INTRODUCTION: Osteochondroma of mandibular condyle is a rare benign tumor. CASE REPORT: This case report described clinical, radiographic features, differential diagnosis, histopathologic correlation and treatment of condylar osteochondroma. CONCLUSION: Conebeam computed tomography (CBCT) is an alternative modality to CT or MRI that should be performed in all cases of suspected osteochondroma of the mandibular condyle.

Maxillary Sinus Floor Augmentation Using Xenograft: Gene Expression and Histologic Analysis.

PURPOSE: Many histologic and histomorphometric studies as well as systematic reviews have shown the clinical success of the use of anorganic bovine bone (ABB, Bio-Oss) in maxillary sinus floor augmentation (MSFA). The molecular processes involved in bone healing are, however, still unknown. The aims of this study were to explore gene expression associated with bone remodeling and inflammation in MSFA sites. MATERIALS AND METHODS: The mRNA expression levels of runt related transcription factor 2 (RUNX2), receptor activator of NF-kB ligand (RANKL), osteoprotegerin (OPG), matrix metallopeptidase 9 (MMP-9), tartrate-resistance acid phosphatase (TRAP), and interleukin-1beta (IL-1ß), as well as the ratio of RANKL/OPG were compared between alveolar bone of a group after MSFA with ABB and a maxillary posterior edentulous bone group. Twenty-one bone samples were collected at the time of implant placement after 6 months of MSFA or tooth extraction. Fourteen bone samples from the MSFA group and from the maxillary posterior edentulous bone without MSFA group were taken to analyze gene expression by real-time reverse transcription polymerase chain reaction (RT-PCR). Seven bone samples from the MSFA group were used for histologic analysis. RESULTS: Real time RT-PCR revealed no statistically significant difference in gene expression level of RUNX2, RANKL, OPG, MMP-9, TRAP, and IL-1ß, or in the ratio of RANKL/OPG. Histology showed bone-lining cells at the edge and osteocyte inside newly formed bone. Residual grafted particles were in close contact with new bone. CONCLUSION: After a healing period of 6 months, ABB particles did not have an effect on the expression of genes associated with bone remodeling and inflammation. In addition, histologic evidence supports that ABB particles are replaced by new bone formation and do not affect bone healing.

Osteoblastic differentiation enhances expression of TRPV4 that is required for calcium oscillation induced by mechanical force.

Mechanical stress is known to alter bone mass and the loss of force stimuli leads to reduction of bone mass. However, molecules involved in this phenomenon are incompletely understood. As mechanical force would affect signaling events in cells, we focused on a calcium channel, TRPV4 regarding its role in the effects of force stimuli on calcium in osteoblasts. TRPV4 expression levels were enhanced upon differentiation of osteoblasts in culture. We found that BMP-2 treatment enhanced TRPV4 gene expression in a dose dependent manner. BMP-2 effects on TRPV4 expression were suppressed by inhibitors for transcription and new protein synthesis. In these osteoblasts, a TRPV4-selective agonist, 4α-PDD, enhanced calcium signaling and the effects of 4α-PDD were enhanced in differentiated osteoblasts compared to the control cells. Fluid flow, as a mechanical stimulation, induced intracellular calcium oscillation in wild type osteoblasts. In contrast, TRPV4 deficiency suppressed calcium oscillation significantly even when the cells were subjected to fluid flow. These data suggest that TRPV4 is involved in the flow-induced calcium signaling in osteoblasts.

Osteopontin deficiency enhances parathyroid hormone/ parathyroid hormone related peptide receptor (PPR) signaling-induced alteration in tooth formation and odontoblastic morphology.

Parathyroid hormone/parathyroid hormone-related protein receptor (PPR) signaling is known to be involved in tooth development. In bone, extracellular matrix protein osteopontin (OPN) is a negative regulator of PPR signaling in bone formation. However, the role of OPN in modulation of PPR action in tooth development is not understood. Therefore, we examined the tooth in double mutant mice. Constitutively active PPR was expressed specifically in the odontoblasts and osteoblasts (caPPR-tg) in the presence or absence of OPN. Radiographic analysis indicated that the length of the third molar (M3) and the incisor was decreased in the caPPR-tg mice compared to wild type, and such reduction in molar and incisor length was further enhanced in the absence of OPN (caPPR-tg OPN-KO). With respect to histology of incisors, caPPR-tg induced high cellularity and irregularity in odontoblastic shape and this was enhanced by the absence of OPN. These morphological observations suggest that OPN modulates PPR signaling that are involved in tooth formation.

Nanogel-based scaffold delivery of prostaglandin E(2) receptor-specific agonist in combination with a low dose of growth factor heals critical-size bone defects in mice.

OBJECTIVE: Regeneration of bone requires the combination of appropriate drugs and an appropriate delivery system to control cell behavior. However, the delivery of multiple drugs to heal bone is complicated by the availability of carriers. The aim of this study was to explore a new system for delivery of a selective EP4 receptor agonist (EP4A) in combination with low-dose bone morphogenetic protein 2 (BMP-2). METHODS: Combined delivery of EP4A and BMP-2 was carried out with a nanogel-based scaffold in the shape of a disc, to repair critical-size circle-shaped bone defects in calvariae that otherwise did not heal spontaneously. RESULTS: Combination treatment with EP4A and low-dose BMP-2 in nanogel efficiently activated bone cells to regenerate calvarial bone by forming both outer and inner cortical plates as well as bone marrow tissue to regenerate a structure similar to that of intact calvaria. EP4A enhanced low-dose BMP-2-induced cell differentiation and activation of transcription events in osteoblasts. CONCLUSION: These data indicate that combined delivery of EP4A and low-dose BMP-2 via nanogel-based hydrogel provides a new system for bone repair.