Objective:?To investigate the effect of magnesium-doped TiO2 coating on titanium by micro-arc oxidation on promoting osteogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs). Methods: The micro arc oxidation (MAO) treatment of titanium surface in calcium and phosphorus electrolyte was set as the control group. By adding different concentrations of magnesium into the electrolyte, Low (Mg-1) and high magnesium (Mg-2) content MAO treated titanium surfaces were constructed.The surface morphology was observed by field emission scanning electron microscope (SEM), the chemical elements on the surface were analyzed by X-ray photoelectron spectroscopy (EDS). MTT method was applied to detect the metabolic activity of BMSCs, cytoskeleton staining was used to observe the growth of BMSCs, alkaline phosphatase (ALP) staining/semi quantitative analysis was used to analyze the expression of ALP protein. The osteogenic differentiation related genes, including Alp,Runx2, Ocn, Bmp2, Bsp, Opn, were analyzed by real-time PCR. The expression of OCN protein was detected by immunofluorescence. Results:BMSCs grew well on the titanium surface of MAO, Mg-1 and Mg-2 groups. The metabolism activity of BMSCs in Mg-2 group was the best (P<0.05). Compared with MAO, the expressions of osteogenic differentiation related genes were increased in both Mg-1 and Mg-2 groups, and the expressions of ALP and OCN protein were enhanced as well. The expression of Runx2, Alp gene expression and ALP protein expression was significantly higher in Mg-1 group when compared with Mg-2 group, while the osteogenic related genes such as Ocn,Bmp2, Bsp,Opnand protein expression of OCN were significantly up-regulated in Mg-2 when compared to Mg-1 group (P<0.05). Conclusions: The incorporation of magnesium ion into the micro-arc oxidation treated titanium surface possessed good biocompatibility, promoted the proliferation of rat BMSCs, significantly improved the osteogenic differentiation of titanium surface, and has a potential in promoting bone implant contact.

Objective:?To investigate the effect of chloromethyl-benzamidodialkylcarbocyanine(CM-Dil) on the proliferation and differentiation of human gingival mesenchymal stem cells(hGMSCs) in vitro and the duration of fluorescence of CM-Dil in vivo. Methods:?hGMSCs were primary cultured by tissue mass method, and surface markers were detected by flow cytometry. After hGMSCs were labeled by CM-Dil, the proliferation was analyzed using CCK-8 assay. Osteogenic differentiation of hGMSCs was investigated using Alizarin Red staining, lipid-forming ability of hGMSCs was detected by Oil Red O staining. The effect of CM-Dil-labeled hGMSCs in vivo was detected in rat skull defect experiment. Results:?The hGMSCs were positive for CD105, CD90 and CD73, but negative for CD14, CD34, CD19, CD45 and HLA-DR. Comparing with the control group, CM-Dil did not affect the proliferation of hGMSCs (P＞0.05), nor did it affect the osteogenic and lipogenic differentiation ability of hGMSCs. In vivo the CM-Dil-labeled hGMSCs could still be observed at week 8. Conclusions:?CM-Dil does not affect the characteristics of hGMSCs in vitro, and the CM-Dil-labeled hGMSCs can be observed for up to 8 weeks in vivo, and CM-Dil can be used as a tracking label of MSC in vivo in the future.

Objective:?To investigate the effect of miR-206-3p on osteoclastogenesis. Methods:?RAW264.7 cells were induced into osteoclasts in vitro. The knockdown and over expression efficiencies of miR-206-3p were examined by real-time fluorescence quantitative PCR (qRT-PCR). Then F-actin staining and trap-resistant acid phosphatase (TRAP) staining were performed to detect the osteoclastogenesis, and the expression of osteoclast differentiation-related gene NFATc1 was evaluated by qRT-PCR and Western Blot. Results:?The overexpression of miR-206-3p significantly inhibited osteoclastogenesis and the expression of NFATc1, while the inhibition of miR-206-3p did the opposite effect (P<0.05). Conclusions:?miR-206-3p regulates the differentiation of RAW264.7 cells into osteoclasts by targeting the expression of NFATc1.

Objective:?To analyze the changes of glycosaminoglycan (GAG) in bone tissue of osteoporosis mice. Methods:?We constructed the animal model of hormone-deficient osteoporosis mice, and assessed whether the model was established successfully by Micro CT and histological section staining. The content of femoral GAG analyzed by Toluidine blue staining and the expression of related gene measured by RT-qPCR were compared between the two groups. Results: Three months after surgery, the experimental mice had an increased total fat and body weight, while the weight of uterus and bone density are decreased. The content of GAG in bone tissue decreased and mRNA expression level of related genes to GAG chain decreased (P<0.05). Conclusion: GAG is closely related to the occurrence and development of osteoporosis.

Objective: To investigate the correlation between M1 macrophages and osteoclastogenesis in periodontitis. Methods: Periodontitis model was established of the right maxillary on 2-month-old C57BL/6J mice, with left maxilla as control. Mice were sacrificed 7 days after surgery. Micro-CT was used to detect alveolar bone loss and histological staining was conducted to examine M1 macrophages and osteoclasts. The expression of genes related to M1 macrophages and osteoclastogenesis was detected by real-time RT-PCR. The correlation between numbers of M1 macrophages and osteoclasts, and location relation of them were analyzed. Results:? Mouse periodontitis model was established successfully. Micro-CT results showed significant decrease of bone volume, BV/TV, Tb.N, Tb.Th and BMD of alveolar bone in right maxilla with periodontitis (P<0.05). And histological staining showed that number of M1 macrophages and osteoclasts significantly increased in maxilla with periodontitis (P<0.05). Expressions of M1 macrophages related genes (Il-1β, Il-6, Tnf-α, iNos), and osteoclastogenesis genes (Oscar, Mmp9, Ctsk, Trap) significantly elevated in periodontitis (P<0.05). Correlation analysis results indicated positive correlation between M1 macrophages and osteoclastogenesis in periodontitis. Co-staining results showed that osteoclast mainly located on alveolar bone surface with M1 macrophages around. Conclusions: In periodontitis, M1 macrophages and osteoclasts were increased and there was positive correlation between them, and there were many M1 macrophages around the alveolar bone surface where osteoclasts located.

Objective:?To investigate the prevalence and distribution of total bacteria, Porphyromonas gingivalis (P. gingivalis), Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans) and leukotoxin (LTX) in Jiangsu Han nationality periodontitits subjects categorized by classifications in 2018 and 1999. Methods:?73 patients with periodontitis and 26 periodontal healthy individuals were recruited. Periodontal clinical parameters were recorded, including probing depth (PD), clinical attachment loss (CAL), gingival index (GI) and plaque index (PLI). Levels of total bacteria, P. gingivalis and A. actinomycetemcomitans in subgingival plaque were explored by real-time PCR, and LTX was detected by conventional PCR. Results:?There were no differences in the levels of total bacteria and P. gingivalis between chronic and aggressive periodontitis (P>0.05), while differences were found when subjects were grouped by staging and grading (P＜0.05). Moreover, differences in the levels of A. actinomycetemcomitans and positive rates of LTX between chronic and aggressive periodontitis patients, between Stage Ⅰ-Ⅱ and Ⅲ-Ⅳ patients, and between Grade A and C patients were disclosed (P>0.05). All LTXs detected in this study were subtypes of low virulence. Positive correlations were confirmed between periodontal clinical parameters (PD, CAL and GI) and the levels of total bacteria, P. gingivalis and A. actinomycetemcomitans. (P＜0.05) Conclusions:?Classification for periodontal diseases in 2018 well reflected the microbiological characteristics of Jiangsu Han nationality periodontitis patients.

Objective:?To study the effects of heme concentration on the pathogenic potential of P. gingivalis in C. albicans - P. gingivalis dual-species biofilm. Methods:?P. gingivalis-C. albicans dual-species biofilm and P. gingivalis single-species biofilm were established in a low- and high- heme environment. The living and dying status of microorganisms in biofilms was observed by laser confocal microscope. Trypsin-like proteinase (TLP) activity of P. gingivalis in culture supernatant and cells was determined by standard curve. The drug resistance of P. gingivalis was detected by K-B disk agar diffusion method. Results:?Under the condition of low heme culture, the number of viable P. gingivalis in the dual-species biofilm was significantly higher than that in the single-species biofilm, and as well as the thickness of the dual-species biofilm was wider in 3D images. The TLP activity in the supernatant and cell of P. gingivalis in dual-species biofilm was higher than that in single-species biofilm (P<0.05). In addition, the resistance rates of P. gingivalis in the dual-species biofilm against cefazolin and ciprofloxacin were also increased, while they were sensitive to sulfamethoxazole. There was no significant differences in Live/Death rate, TLP activity and drug sensitivity between the dual- and single-species biofilm under high heme culture condition, but the TLP activity and the resistance against cefazolin and ciprofloxacin were lower than those of the biofilms cultured in a low-heme environment. Conclusions:?Under the condition of low heme culture, the pathogenicity potential of P. gingivalis in the dual-species biofilm was increased.

Objective:?Screening for differentially expressed genes between inflammatory pulp and healthy pulp. Methods:? Transcriptome RNA sequencing was performed on 5 inflammatory dental pulp samples and 6 healthy dental pulp samples. Differentially expressed genes were screened for bioinformatics analysis. Results:? A total of 1 216 differentially expressed genes were screened out, including 831 up-regulated genes and 385 down-regulated genes. GO analysis revealed that the differentially expressed genes were concentrated in the functional sets consisting of inflammatory response, immune response and cell adhesion. KEGG pathway analysis showed that the differentially expressed genes were enriched in inflammatory pathways such as cytokine-cytokine receptor interactions, chemokine signaling pathways and osteoclast differentiation. GSEA results were also enriched in the inflammatory response pathway, TNF-α signaling pathway via NF-κB, and IL6/JAK/STAT3 signaling pathway. According to the degrees of nodes in the PPI network, 6 hub genes related to pulpitis were screened out: PTPRC, ITGAM, CD86, IL10, TLR4 and TLR2. Conclusions:? There were significantly statistical differences in the overall gene expression between inflammatory and healthy pulp. Most of the differentially expressed genes are concentrated in the functions and signaling pathways of inflammation and immune response, and the related genes have potential value as biomarkers for the diagnosis of pulpitis.

Objective：3D printing technology combined with emulsion template to manufacture PLGA/β-tricalcium phosphate(β- TCP)composite scaffolds(PT)with different proportions, to screen out the most significant group, and to evaluate its potential to repair large bone defects in maxillofacial region. Methods：PLGA was dissolved in dichloromethane and made into water-in-oil emulsion. β-TCP was added in proportion and mixed evenly to form scaffolds by 3D printing. The scaffolds were recorded as PT30, PT50, PT70 according to the content of TCP. After that, a series of structural characterization, physical properties and in vitro biocompatibility tests were carried out on the scaffolds. Results：With the increase of TCP content, the surface roughness of the scaffold increased, the pore size and porosity were not affected (P > 0.05), the water absorption rate increased significantly (P < 0.001), and the mechanical properties were not significantly different (P > 0.05). Only the weight and pH of PT30 scaffold decreased most significantly (P < 0.05) during degradation, and there was no significant difference between the other two groups (P > 0.05). PT30 and PT50 scaffolds have no cytotoxicity, while PT70 scaffolds are screened out because of their cytotoxicity. In addition, the number of cell adhesion increased with the increase of TCP content, and PT30 scaffold was also screened out. Conclusions：PT50 scaffold had good water absorption and mechanical properties, and its degradation speed is suitable for new bone growth, it had no cytotoxicity and was suitable for cell growth and adhesion.

To preliminarily evaluate the clinical efficacy of immediate implantation and immediate restoration on patients with potential edentulous jaws. Methods:?A retrospective analysis of 17 patients with poetential edentulous jaws from January 2016 to December 2019 was conducted, including 4 patients in the upper jaw, 7 patients in the lower jaw, and 6 patients in both jaws. Immediate implantation and immediate restoration were adopted. After 24 months (range from 12 to 36 months) of the mean follow-up period, the clinical efficacy was analyzed. Results:?All patients have successfully completed the entire treatment procedure. A total of 128 implants were implanted in 17 patients. No implant osseointegration failure occurred during the observation period. Two cases had peri-implant mucositis after operation while 4 cases had mechanical complications of temporary restorations, and 3 cases had mechanical complications of permanent restorations, all of which were porcelain /plastic fracture. All patients were satisfied with the chewing and aesthetic effects. Conclusions:?Patients with poetential edentulous jaws undergo immediate implantation and immediate restoration can achieve ideal clinical effect under the conditions of strictly grasping the indications, ensuring the implant torque, and obtaining the cooperation of patients.

Objective: To evaluate the application value of near infrared fluorescence navigation technology in oral cancer surgery and its surgical margin. Methods: Fourteen patients with oral cancer who received surgical treatment in the Stomatological Hospital of Nanjing Medical University were selected in this study. Each patient received 0.75?mg/Kg indocyanine green injection intravenously 6-8h before surgery. Near infrared fluorescence imaging system was used to observe and record during and after operation. Specimens were collected within and outside the fluorescence boundary of the isolated lesions for pathological examination after surgery. Results: Fluorescence imaging was observed in all the lesions, and the tumor background ratio (1.58±0.52) was significantly higher than that of the intraoperative lesions (1.39±0.39) (P<0.05). At the fluorescence boundary, the sensitivity and specificity of differentiating normal epithelium from abnormal epithelium (dysplasia and cancer tissues) were 83.3% and 57.9% respectively. Conclusions: NIR fluorescence technique can be used to detect the residual tumor after the resection of the primary oral cancer, and it is expected to be an auxiliary tool to assist clinicians in determining the surgical margin.

Maintaining the stability of cell volume is an important condition in the process of cell life. Volume regulated anion channel (VRAC) plays an important role in regulating the osmotic pressure inside and outside the cell membrane through organic matter accumulation and inorganic ion exchange. VRAC regulates cell volume and participates in a series of biological and cellular activities such as nervous system, immune system, metabolism, cell migration, proliferation and apoptosis. Recently, studies have confirmed that leucine rich repeat containing protein 8A (IRRC8a, also known as Swell1) and its four homologous family members are an important VRAC and play an important role in the regulation of cell volume. In this paper, we reviewed the structure of Swell1,as well as its effect and mechanism on different cell volume regulation.

Periodontal ligament stem cells (PDLSCs) used in tissue engineering is one of the most promising methods for the treatment of periodontitis with bone loss. However, the effect of PDLSCs in vivo for periodontal tissue regeneration is not ideal, which may be due to the influence of the inflammatory microenvironment of PDLSCs on their fate. Therefore, this review will explore the effect of inflammatory microenvironment on the osteogenic differentiation potential of PDLSCs from the aspect of signal pathway, so as to provide theoretical basis for the prevention and treatment strategy of bone loss in periodontitis.