Therapeutic Effect of a Novel M1 Macrophage-Targeted Nanodrug in Chronic Periodontitis Mice.

Chronic periodontitis is a chronic, progressive, and destructive disease. Especially, the large accumulation of advanced glycation end products (AGEs) in a diseased body will aggravate the periodontal tissue damage, and AGEs induce M1 macrophages. In this project, the novel nanodrugs, glucose-PEG-PLGA@MCC950 (GLU@MCC), are designed to achieve active targeting with the help of glucose transporter 1 (GLUT1) which is highly expressed in M1 macrophages induced by AGEs. Then, the nanodrugs release MCC950, which is a kind of NLRP3 inhibitor. These nanodrugs not only can improve the water solubility of MCC950 but also exhibit superior characteristics, such as small size, stability, innocuity, etc. In vivo experiments showed that GLU@MCC could reduce periodontal tissue damage and inhibit cell apoptosis in periodontitis model mice. In vitro experiments verified that its mechanism of action might be closely related to the inhibition of the NLRP3 inflammatory factor in M1 macrophages. GLU@MCC could effectively reduce the damage to H400 cells caused by AGEs, decrease the expression of NLRP3, and also obviously reduce the M1-type macrophage pro-inflammatory factors such as IL-18, IL-1ß, caspase-1, and TNF-α. Meanwhile, the expression of anti-inflammatory factor Arg-1 in the M2 macrophage was increased. In brief, GLU@MCC would inhibit the expression of inflammatory factor NLRP3 and exert antiperiodontal tissue damage in chronic periodontitis via GLUT1 in the M1 macrophage as the gating target. This study provides a novel nanodrug for chronic periodontitis treatment.

Interleukin-37 ameliorates periodontitis development by inhibiting NLRP3 inflammasome activation and modulating M1/M2 macrophage polarization.

OBJECTIVE: Our study was designed to explore the role of IL-37 in M1/M2 macrophage polarization imbalance in the pathogenesis of periodontitis. BACKGROUND: Periodontitis is a chronic progressive inflammatory disease featured by gingival inflammation and alveolar bone resorption. Recent research has revealed that regulating macrophage polarization is a viable method to ameliorate periodontal inflammation. IL-37 is an anti-inflammatory cytokine, which has been reported to inhibit innate and adaptive immunity. METHODS: For in vitro experiment, mouse macrophage RAW264.7 cells were pretreated with 0.1 ng/mL recombinant human IL-37. M1 and M2 polarizations of RAW264.7 cells were induced by 100 ng/mL LPS and 20 ng/mL IL-4, respectively. The expression of M1 (iNOS, TNF-α, and IL-6) and M2 (CD206, Arg1, and IL-10) phenotype markers in RAW264.7 cells was detected by RT-qPCR, western blotting, and immunofluorescence staining. For in vivo experiment, experimental periodontitis mouse models were established by sterile silk ligation (5-0) around the bilateral maxillary second molar of mice for 1 week. H&E staining of the maxillary alveolar bone was used to show the resorption of root cementum and dentin. Alveolar bone loss in mouse models was evaluated through micro-CT analysis. The expression of iNOS and CD206 in gingival tissues was assessed by immunohistochemistry staining. NLRP3 inflammasome activation was confirmed by western blotting. RESULTS: IL-37 pretreatment reduced iNOS, TNF-α, and IL-6 expression in LPS-treated RAW264.7 cells but increased CD206, Arg1, and IL-10 in IL-4-treated RAW264.7 cells. LPS-induced upregulation in NLRP3, GSDMD, cleaved-IL-1ß, and cleaved-caspase-1 expression was antagonized by IL-37 treatment. In addition, IL-37 administration ameliorated the resorption of root cementum and dentin in periodontitis mouse models. IL-37 prominently decreased iNOS+ cell population but increased CD206+ cell population in gingival tissues of periodontitis mice. The enhancement in NLRP3, GSDMD, cleaved-IL-1ß, and cleaved-caspase-1 expression in the gingival tissues of periodontitis mice was offset by IL-37 administration. CONCLUSION: IL-37 prevents the progression of periodontitis by suppressing NLRP3 inflammasome activation and mediating M1/M2 macrophage polarization.

Quantitative and qualitative condylar changes following stabilization splint therapy in patients with temporomandibular joint disorders with and without skeletal lateral mandibular asymmetry: a cone beam computed tomographic study.

BACKGROUND: Temporomandibular disorders (TMDs) encompass pain and dysfunction in the jaw, muscles, and adjacent structures. This study aimed to explore the quantitative (condylar position, morphology) and qualitative (bone mineral density (BMD)) therapeutic outcomes following a stabilization splint (S.S.) therapy in adult patients diagnosed with TMD (Arthralgia) with/without lateral mandibular asymmetry (MA) using cone beam computed tomography (CBCT). METHODS: In this retrospective clinical study, 60 adult TMD patients who received S.S. therapy were enrolled and allocated into the TMD group (TMDG) and TMD with MA group (TMD + MAG). The diagnosis was made according to the Diagnostic Criteria for TMD (DC/TMD) AXIS I. MA was measured from the mid-sagittal plane to the Menton point. CBCT was used to scan the temporomandibular joints pre- (T0) and post- (T1)-treatment for three-dimensional analysis. Intra- and intergroup statistical comparisons were performed using the Wilcoxon signed ranks and the Kruskal‒Wallis test. RESULTS: For quantitative comparisons, there was a statistically significant difference between T0 and T1 in the joint spaces of TMD + MAG (anterior, superior, posterior, and coronal lateral on the deviated side as well as in the superior, coronal medial joint space of the contralateral side). Morphologically, the deviated side had a narrower condylar width, reduced condylar height, and a steeper eminence angle. In contrast, the contralateral side tended to have a greater condylar length. For qualitative measurements, BMD also showed statistical significance between T0 and T1 in the majority of the condyle slopes (AS, SS, PS, and LS on the deviated side and in AS and MS on the contralateral side) of TMD + MAG. Additionally, only the AS and PS showed significance in TMDG. CONCLUSION: Multiple joint space widening (AJS and CMS) and narrowing (SJS, PJS, and CLS) could characterize the deviated side in TMD + MA. Factors like narrower condylar width, reduced condylar height, and steeper eminence angle on the deviated side can worsen TMD + MA. Proper alignment of the condyle-disc position is essential for optimal function and load distribution, potentially affecting bone mineral density (BMD). MA plays a prominent role in disturbing bone densities. S.S. therapy shows more evident outcomes in TMD + MAG (on the deviated side compared to the contralateral side) than the TMDG.

Skeletal, dentoalveolar and soft tissue changes after stabilization splint treatment for patients with temporomandibular joint disorders.

BACKGROUND: Temporomandibular disorder (TMD) is a grouping of heterogeneous disorders with multifactorial origins. Stabilization splints (SS) have demonstrated an acceptable treatment effect in TMD. The possible changes at the skeletal, dental, and soft tissue levels need to be addressed to evaluate the benefit/risk ratio of this therapeutic procedure. Accordingly, this study aimed to three­dimensionally evaluate skeletal, dentoalveolar and soft tissue changes after SS treatment for patients with TMD. METHODS: This retrospective study included 74 adult patients with myofascial and/or intra-articular disorders (25 males and 49 females), with an average age of 22.88 ± 4.8 years, who underwent SS treatment. Pre- and post-treatment Cone beam computed tomography were analysed using Invivo 6.0.3 software. The primary outcome was the vertical skeletal and dentoalveolar changes, while the secondary outcomes were the anteroposterior skeletal, dentoalveolar and soft tissue changes. Paired t-test and Wilcoxon rank sum test were used for statistical analyses. RESULTS: For the primary outcome; skeletally, there was a significant increase in mandibular plane inclination (difference: 0.82°±1.37), decrease facial height ratio (difference: 0.45%±1.07) and at the dentoalveolar level, the inclination of the functional (FOP-SN, FOP-FH) and bisecting (BOP-SN, BOP-FH) occlusal planes exhibited a significant increase too (difference: 0.38 ± 1.43°, 0.49 ± 1.62°, 0.44 ± 1.29° and 0.41 ± 1.17°, respectively) and also a decrease in the overbite (difference: -0.54 ± 0.83). For the secondary outcomes; there was a significant decrease in mandibular position (SNB) (difference: 1.60 ± 1.36°) and increase in the overjet (difference: 0.93 ± 1.04, p < 0.001) and a significant lower lip retrusion (difference: 0.33 ± 1.01 mm p < 0.01), was observed too. CONCLUSIONS: SS therapy resulted in significant vertical skeletal and dentoalveolar changes that were manifested mainly by facial height ratio, mandibular and occlusal plane changes, and to a lesser extent, significant anteroposterior skeletal, dentoalveolar, and soft tissue changes in the form of mandibular position, increased overjet and a more retrusive lower lip. These changes should be considered during patients' selection prior to initiating SS therapy.

Influence of polyether ether ketone coping crown on the adaptation of implant abutment.

OBJECTIVE: To evaluate the influence of polyether ether ketone coping crown on the adaptation of implant abutment. Methods: The vitro study was conducted at the department of Prosthodontics, Hainan Stomatological Hospital, China, from October 2021 to March 2022, and comprised patients undergoing implant surgery on first molar. Patients were divided into two groups, with group A patients receiving polyether ether ketone coping crowns, and group B receiving zirconia crowns. Replica technique was used to replicate the gap between the crowns and the abutments. The thickness of the silicone film was measured under the stereomicroscope, and the gap between the groups was compared. Data was analysed using SPSS 22. RESULTS: In group A, mean marginal gap was 82.43±25.00µm, and mean overall gap was 85.45±33.75µm. In group B, the corresponding values were 65.09±11.69µm and 78.04±26.67µm. There was a significant difference in the adaptation between the groups at the marginal and overall measurement points (p<0.05). Conclusion: Marginal and internal adaptations of polyether ether ketone coping crown for abutment could be considered clinically acceptable.

Quantitative and qualitative condylar changes following stabilization splint therapy in patients with temporomandibular joint disorders.

OBJECTIVE: This study aimed to explore the quantitative and qualitative condylar changes following stabilization splint (S.S) therapy, including condylar position, morphology, and bone mineral density (BMD) in subjects with temporomandibular disorders (TMD). MATERIALS AND METHODS: In this retrospective clinical study, we enrolled 40 TMD subjects (80 joints) aged 18 to 35 years, for whom a S.S was used to treat TMD. The 80 TMD consists of 32 masticatory muscle disorders (myalgia) and 48 TMJ disorders (arthralgia). Cone beam computed tomography (CBCT) was used to scan the TMJs of subjects pre- and post-treatment for three-dimensional analysis (3D). Using Mimics software v.21.0, quantitative (3D condylar and joint spaces dimensions parameters were measured using linear measurements in millimeters, according to the Kamelchuk method and Ikeda method, while the assessment of anteroposterior condyle position within the glenoid fossa was based on the method of Pullinger and Hollender), and qualitative (a round bone tissue with an area of 2 mm2 in three representative areas according to the Kamelchuk method to measure condylar BMD) pre- and post-treatment. Intra- and inter-group statistical comparisons were performed using the Wilcoxon signed ranks and the Kruskal-Wallis test, respectively. RESULTS: The course of treatment was 6-12 months, with an average of 9.1 months. For the pre- and post-treatment quantitative comparisons, there was a statistically significant difference in the anterior joint space (AJS) and coronal medial space, as well as the condyle length in the myalgia group and condylar width in the arthralgia group. For qualitative measurements, a significant difference was observed in the posterior slope of the myalgia group and the arthralgia group's anterior, superior, and posterior slopes. The inter-group comparisons revealed significant differences in AJS, condylar length, and anterior slope density. CONCLUSION: In short-term follow-up, the S.S influenced patients with TMD from different origins; it changes anterior and coronal medial joint space, condyle length in myalgia, and width in arthralgia. Furthermore, it improved the condyle bone density more evidently in arthralgia. CLINICAL RELEVANCE: This study highlights the influence of S.S on symptomatic populations with TMD of different origins from a qualitative and quantitative perspective.

Efficacy of mandibular molar distalization by clear aligner treatment. / æ— æ‰˜æ§½éšå½¢çŸ«æ²»å™¨è¿œç§»ä¸‹é¢Œç£¨ç‰™çš„ç–—æ•ˆ.

OBJECTIVES: At present, the research on clear aligner of molar distalization mainly focuses on the upper jaw, while the research on mandibular molars is few.This study aims to evaluate the therapeutic effect of mandibular molars distalization with clear aligner via cone beam CT (CBCT) and Dolphin software. METHODS: Twenty cases of mandibular molars with clear aligner were included according to the inclusion and exclusion criteria. CBCT was taken before treatment (T0) and when the first molar was moved in place (T1). Dolphin software was used to measure the effectiveness of molar distalization. Three-dimensional changes in direction and the impact on the incisors and facial soft and hard tissues were evaluated. RESULTS: The effective rates of crown and root distalization of the second and first mandibular molars were 74%, 49%, and 71%, 47%, respectively. The second and first molars were both the distal buccal cusp with the largest distalization [(2.15 ± 0.91) mm and (1.85±1.09) mm], respectively, with significant difference between the T0 and T1 (P<0.05). The second and first molars were accompanied by depression, distal tilt, and buccal tilt with 1.06 mm, 2.10°, 2.27°, and 0.91 mm, 1.62°, and 1.91°, respectively, with significant differences between the T0 and T1 (all P<0.05). There was no obvious difference between men and women. The mandibular central incisor showed a lip-side movement of 1.02 mm, a depression of 0.82 mm, a mesial incline of 0.66°, and a crown-lip torque of 1.51° after molar distalization, with significant differences between the T0 and T1 (all P<0.001). Only the lower lip thickness increased by 0.1 cm, the length of the lower lip increased by 0.1 cm, and the ANS-ME (distance from anterior nasal spine to submental point) decreased by 0.13 cm, with significant differences between the T0 and T1 (all P<0.05). CONCLUSIONS: Clear aligner can effectively move mandibular molars farther, the crown is more effective than the root, and it is tilted. The second mandibular molar is more effective than the first mandibular molar in its distant displacement and three-dimensional changes. Molar distalization causes minor changes in mandibular incisors and facial soft and hard tissues.

Three-dimensional evaluation of upper pharyngeal airway, hyoid bone, and craniocervical changes following stabilization splint therapy in adult patients with temporomandibular joint disorders and mandibular deviation: A retrospective study.

OBJECTIVE: This study aimed primarily to analyze the three-dimensional (3D) changes in the pharyngeal airway (PA), and secondarily, the hyoid bone (HB) and the craniocervical (CC) following stabilization splint (SS) therapy in adult patients with temporomandibular joint disorders (TMD) and mandibular deviation (MD). METHODS: Thirty-five adult patients with TMD and MD, who were treated using SS with a mean age of 25.14 ± 6.11 years, were enrolled in this retrospective clinical study. Pre- and post-therapeutic cone-beam computed tomography (CBCT) scans were analyzed. PA dimension,nasopharyngeal, oropharyngeal, hypopharyngeal, sub-hypopharyngeal, and total pharyngeal airway spaces were measured in surface area, volume, minimum constricted area (MCA) and width, HB position, and CC posture were analyzed three-dimensionally using InVivo 6.0.3 and Dolphin 11.95 software. Wilcoxon rank-sum or Paired t-test was conducted, and P < 0.05 was considered significant. RESULTS: SS therapy was administered for a period of 9.49 ± 4.02 months. The oropharyngeal airway space showed a significant decrease in sagittal width. The hypopharyngeal surface area, volume, MCA, and sagittal width decreased significantly. In terms of HB, hyoid-mandibular plane (H-MP), retrognathia-third vertebra's most inferior-anterior (RGN-C3ia), and retrognathia-Sella (RGN-S) distances significantly decreased. The Nasion-Sella line and the line that passes through C2ip to the odontoid process posterior tangent (NSL-OPT) angle in CC posture also decreased significantly. CONCLUSION: SS therapy in TMD patients with MD mainly results in narrowing of the hypopharyngeal region, no change in HB position and improvement in head posture. These results undoubtedly assist in diagnosis and treatment of clinical conditions.

Effect of Tensile Frequency on the Osteogenic Differentiation of Periodontal Ligament Stem Cells.

Purpose: The role of periodontal ligament stem cells (PDLSCs) in mediating osteogenesis involved in orthodontic tooth movement (OTM) is well established. However, various relevant in vitro studies vary in the frequency of tension. The effect of tensile frequency on the mechanotransduction of PDLSCs is not clear. The current study aimed to determine the effect of different tensile frequencies on the osteogenic differentiation of PDLSCs and to identify important mechano-sensitivity genes. Methods: Human PDLSCs were isolated, identified, and subjected to cyclic equibiaxial tensile strain of 12% at different frequencies of 0.1 Hz, 0.5 Hz, 0.7 Hz, or static cultures. Osteogenic differentiation of PDLSCs was assessed by using Western blotting. High-throughput sequencing was used to identify differential mRNA expression. Short time-series expression miner (STEM) was utilized to describe the frequency patterns of the mRNAs. The functions and enriched pathways were identified, and the hub genes were identified and validated. Results: We found that the osteoblastic differentiation capacity of PDLSCs increased with tensile frequency in the range of 0.1-0.7 Hz. Eight frequency-tendency gene expression profiles were identified to be statistically significant. Tensile frequency-specific expressed genes, such as SALL1 and EYA1, which decreased with the increase in tensile frequency, were found. Conclusion: The osteoblastic differentiation of PDLSCs under mechanical tensile force is frequency dependent. EYA1 and SALL1 were identified as potential important tensile frequency-sensitive genes, which may contribute to the cyclic tension-induced osteogenic differentiation of PDLSCs in a frequency-dependent manner.

Three-Dimensional Culture Promotes the Differentiation of Human Dental Pulp Mesenchymal Stem Cells Into Insulin-Producing Cells for Improving the Diabetes Therapy.

INTRODUCTION: Diabetes is a metabolic disease with a high incidence and serious harm to human health. Islet ß-cell function defects can occur in the late stage of type 1 diabetes and type 2 diabetes. Studies have shown that stem cell is a promising new approach in bioengineering regenerative medicine. In the study of stem cell differentiation, three-dimensional (3D) cell culture is more capable of mimicking the microenvironment of cell growth in vivo than two-dimensional (2D) cell culture. The natural contact between cells and cells, and cells and extracellular matrix can regulate the development process and promote the formation of the artificial regenerative organs and organization. Type IV, VI collagen and laminin are the most abundant extracellular matrix components in islets. Matrigel, a basement membrane matrix biomaterial rich in laminin and collagen IV. MATERIALS AND METHODS: We used Matrigel biomaterial to physically embed human dental pulp stem cells (hDPSCs) to provide vector and 3D culture conditions for cells, and we explored and compared the preparation methods and preliminary mechanisms of differentiation of hDPSCs into insulin-producing cells (IPCs) under 2D or 3D culture conditions.We first designed and screened the strategy by mimicking the critical events of pancreatogenesis in vivo, and succeeded in establishing a new method for obtaining IPCs from hDPSCs. Activin A, Noggin, and small molecule compounds were used to synergistically induce hDPSCs to differentiate into definitive endoderm-like cells, pancreatic progenitor like cells and IPCs step by step under 2D culture conditions. Then, we used Matrigel to simulate the microenvironment in vivo, induced hDPSCs to differentiate into IPCs in Matrigel, evaluated and compared the efficiency between 2D and 3D culture conditions. RESULTS: The results showed that the synergistic combination of growth factors and small molecule compounds and 3D culture promoted the differentiation of hDPSCs into IPCs, significantly enhancing the release of insulin and C-peptide from IPCs. DISCUSSION: Significant support is provided for obtaining a large number of functional IPCs for disease modeling and final cell therapy in regenerative medicine.

In vivo generation of thick, vascularized hepatic tissue from collagen hydrogel-based hepatic units.

In vivo engineering of hepatic tissue based on primary hepatocytes offers new perspectives for the treatment of liver diseases. However, generation of thick, three-dimensional liver tissue has been limited by the lack of vasculature in the tissue-engineered constructs. Here, we used collagen hydrogel as a matrix to generate engineered hepatic units to reconstitute three-dimensional, vascularized hepatic tissue in vivo. Hepatocytes harvested from Sprague-Dawley rats were mixed with liquid type I collagen, concentrated Dulbecco's modified Eagle's medium (2 x), and hepatocyte maintenance medium to create hepatocyte/collagen hydrogel constructs. The constructs were then dissociated into cylindrical hepatic units (diameter/height: 2000-4000 microm/500-1000 microm). Stacking of hepatic units under the subcutaneous space resulted in significant cell engraftment, with the formation of large fused hepatic system (more than 0.5 cm thickness) containing blood vessels. In contrast, only less cell engraftment could be achieved when hepatocytes were transplanted in a manner of whole constructs. Functional maintenance of the engineered hepatic tissue was confirmed by the expression of liver-specific mRNA and proteins. The engineered hepatic tissue has the ability to respond to the regenerative stimulus. In conclusion, large hepatic tissue containing blood vessels could be engineered in vivo by merging small hepatic units. This approach for tissue engineering is simple and represents an efficient way to engineer hepatic tissue in vivo.

Enrichment of cardiomyocytes derived from mouse embryonic stem cells.

BACKGROUND: Embryonic stem (ES) cell-derived cardiomyocytes transplantation and tissue engineering together represent a promising approach for the treatment of myocardial infarction, despite the limited supply of cardiac myocytes. This study examines whether functional cardiomyocytes can be efficiently enriched from mouse embryonic stem (mES) cells. METHODS: mES cells were induced by ascorbic acid to differentiate into cardiomyocytes. Beating cells were observed after 1 week and increased in number with time while under differentiation conditions. Furthermore, the differentiated cultures could be dissociated and enriched by Percoll gradient density centrifugation. RESULTS: The beating cells expressed markers characteristic of cardiomyocytes, such as cardiac troponin T (cTnT). The enriched population contained 88.7% cardiomyocytes and showed expression of cardiomyocyte markers of troponin T and cardiac genes, including alpha-MHC, beta-MHC, ANF and Nkx2.5. However, Oct-4, a marker of early-stage ES cells, was not expressed in the mES cell-derived cardiac cell clusters. Moreover, the mES cell-derived and Percoll-enriched cardiomyocytes responded appropriately to cardioactive drugs, as did normal neonatal rat cardiomyocytes. CONCLUSIONS: mES cell-derived functional cardiomyocytes can be enriched by the method of discontinuous Percoll gradient centrifugation. The ability to differentiate and enrich for functional mouse cardiomyocytes makes it possible for further development of these cells as a model of myocardial repair through cell transplantation or tissue engineering.