上海口腔医学最新文献

Purpose: To clarify the effect of genistein(GEN) on osteogenic differentiation and explore the effect of GEN loaded by platelet-rich fibrin (PRF) on the repair process of bone defects in obese mice.

Methods: In in vitro experiments, the effect of GEN(0, 0.1, 1, 10, 50 μmol/L) on the proliferation of mouse embryonic osteoblast precursor cells (MC3T3-E1) was determined by CCK 8. Alkaline phosphatase(ALP) staining and quantitative detection of ALP activity were performed to determine the changes of ALP activity in cells; RNA and protein expression levels of ALP, osteopontin (OPN) and osteocalcin (OCN) were detected by quantitative real-time PCR(qRT-PCR) and Western blot. Alizarin red staining was used to define the effect of GEN on mineralization of MC3T3-E1. To verify the feasibility of the PRF drug loading, the ultrastructure of PRF was subsequently observed under SEM. In in vivo experiments, obese C57 mouse models were established by high-fat diet feeding. On this basis, skull defect models with a diameter of 2.8 mm were established, and the prepared GEN/PRF complexes were placed into the bone defect area. The effects of GEN on skull defect repair in obese mice were evaluated by Micro-CT scanning and hematoxylin-eosin(H-E) staining. Statistical analysis was performed with GraphPad Prism 5.0 software package.

Results: CCK 8 results showed that 0.1, 1 μmol/L GEN promoted cell proliferation within 7 days(P＜0.05); 10 μmol/L GEN had no significant effect on the process of cell proliferation. From the second day, 50 μmol/L GEN significantly inhibited cell growth and showed cytotoxicity(P＜0.05). These two concentrations had similar effects in promoting cellular osteogenic differentiation. SEM results showed that PRF presented a 3-dimensional network structure, providing space for loading drug molecules. In in vivo experiments, the body weight of mice in the high-fat diet (HFD) group was 27.7% greater than that in the normal diet group(P＜0.05) and had abnormal glucose tolerance (P＜0.05). Micro-CT showed that compared with the normal diet group, the number of bone trabeculae in the femur of obese mice was decreased(P＜0.05), the distance between bone trabeculae was widened(P＜0.05), and the bone density was decreased (P＜0.05). In addition, GEN (0.1, 1.0 μmol/L) loaded by PRF increased bone volume fraction in the skull of obese mice (P＜0.05). H-E results showed that GEN/PRF promoted the healing of the bone defects.

Conclusions: GEN promotes osteogenic differentiation of MC3T3-E1, and it can effectively accelerate the healing of cranial bone defects after loading with PRF in obese mice.

Purpose: To explore the cytotoxic effect of a menthol-favored E-liquid on human periodontal ligament stem cells (hPDLSCs), as well as the underlying mechanism of electronic cigarette (E-cig)-induced cell apoptosis.

Methods: PDLSCs were isolated and cultured from periodontal ligament tissues of healthy premolars extracted for orthodontic reasons. Cells in passage 3 were used to detect the surface markers of stem cells by flow cytometry. Then the cells were exposed to different doses of menthol-favored E-liquid (at 59 mg/L nicotine concentration) in the culture median (the final nicotine concentrations were 0.1 μg/mL, 1.0 μg/mL, 10 μg/mL, 50 μg/mL, 0.1 mg/mL, 0.2 mg/mL and 0.5 mg/mL, respectively) for different period of times (24, 48 and 72 h). The cell viability was analyzed by CCK-8 assay. Cell apoptosis was evaluated by flow cytometry (7-AAD and Annexin V staining) and TUNEL assay. Reactive oxygen species (ROS) production was detected with fluorescence probe DCFH-DA by confocal microscopy and flow cytometry. The protein expression levels associated with ROS/JNK/caspase 3 axis(p-JNK, JNK, c-Jun, p-c-Jun, Bcl-2, Bax and cleaved-caspase 3) were analyzed by Western blot. Immunocytofluorescense staining was applied to evaluate the expression level of p-JNK. After addition of NAC, a ROS scavenger, and MAPK/JNK specific blocker SP600125, their effects on E-cig-induced cell apoptosis were evaluated. Statistical analysis was performed with Graph Pad 5.0 software package.

Results: Human PDLSCs were successfully isolated and cultured and flow cytometry assay showed the mesenchymal stem cell surface biomarkers (CD73, CD90 and CD105) were positively expressed. CCK8 assay indicated cell viability was significantly(P＜0.001) different among all concentration groups at various time points (24, 48 or 72 h), and the difference in apoptosis rate among all concentration groups was also statistically significant (P＜0.001). After exposure to E-liquid with nicotine concentration ≥50 μg/mL, cell viability was significantly reduced, and the proportion of apoptotic cells and the cellular ROS level was significantly increased in a dose-dependent manner as compared with the control group(0.0 mg/mL). Western blot assay showed E-cig exposure could promote MAPK/JNK phosphorylation in a dose-dependent and time-dependent manner. Either NAC or SP600125 could partially rescue the E-cig-induced cell apoptosis via reversing up-regulation of p-JNK and cleaved caspase 3.

Conclusions: ROS/JNK/caspase 3 axis is involved in menthol-favored E-liquid-induced apoptosis of hPDLSCs.

Purpose: To explore the effect of extracting the completely impacted teeth by minimally invasive surgery with preserving the buccal bone plate.

Methods: Eighty-six cases were selected and randomly divided into 2 groups. In the experimental group, a fenestration was made with a ball drill to expose the buccal and lingual margin of crown, and the buccal bone plate was preserved. T-shaped crown cuttings were performed, minimally invasive extraction was conducted.In the control group, the distal and buccal bone plates were removed with a ball drill, the distal and buccal crowns were exposed, and T-shaped crown was cut. The other procedures were the same. The degree of swelling, restricted mouth opening and VAS pain score after operation were observed, the levels of C-reactive protein and anti-hemolytic streptoglobulin were detected by laboratory tests, and the periodontal probing depth(PD), bleeding index (BI), and clinical attachment loss(CAL) of the adjacent second molar were examined 1 month after surgery. SPSS 25.0 software package was used for data analysis.

Results: The swelling degree of the two groups was significantly relieved in the experimental group than in the control group (P＜0.05), and there was no significant difference in the degree of mouth opening limitation and pain (P＞0.05). The level of C-reactive protein in the control group was significantly higher than that in the observation group (P＜0.05). There was no significantly difference in the level of anti-hemolytic streptococcus between the 2 groups (P＞0.05). One month after operation, the PD and CAL in the control group were significantly higher than those in the experimental group(P＜0.05). There was no significant difference between the 2 groups in BI(P＞0.05).

Conclusions: The patients who preserve the buccal bone plate by minimally invasive extraction of impacted mandibular teeth have less reaction and better wound healing.

Purpose: To study the structural characteristics of oral microorganisms in children with caries by 16S rRNA high-throughput sequencing technology.

Methods: Thirty healthy children aged 3-5 years were enrolled as subjects. According to the index of dmfs, they were divided into caries-free (CF) group (15) and early childhood caries (ECC) group(15). To compare the differences in bacterial community structure, samples of saliva and dental plaque were collected, and high-throughput sequencing was conducted using the Illumina Miseq sequencing platform. Bioinformatics analysis was used to analyze the difference of microbial community structure and diversity with SPSS 23.0 software package.

Results: Microbial diversity in ECC group was significantly lower than CF group. At phylum level, Actinobateria was more abundant in saliva samples of ECC group, while Firmicutes was more abundant in plaque samples of CF group. At genus level, the abundance of Lautropia of CF group was higher in saliva samples while Cardiobacterium, Gemella and Granulicatella were abundant in plaque samples. The abundance of Rothia of ECC group was higher in saliva samples and Corynebacterium was abundant of ECC group in plaque samples.

Conclusions: There are significant differences in the species and composition of microbial community in saliva and plaque of children with or without caries. Specific microorganisms are related to the occurrence of ECC, and screening specific microorganisms is helpful for early prediction and prevention of ECC.

Purpose: To investigate the role and mechanism of connexin 43（Cx43）in odontoblast differentiation of human dental pulp cells (hDPCs) induced by lipopolysaccharide (LPS).

Methods: The maxillary first molar injury model of SD rats was established. The expression pattern of Cx43 in dental pulp repair after injury was detected by immunofluorescence(IF) staining. hDPCs was respectively stimulated with 0, 1, 10, 100 and 1 000 ng/mL LPS for 6 h to screen the optimal concentration, and then the expression of Cx43 was inhibited and overexpressed in hDPCs. Quantitative real-time PCR(qRT-PCR) and Western blot(WB) were used to detect the expression of Cx43 and dentin sialophosphoprotein (DSPP), dental matrix protein-1 (DMP-1), osterix (Osx) and extracellular signal-regulated kinase (ERK) activity. Furthermore, hDPCs were treated with specific Cx43 channel inhibitors to investigate the effect of Cx43-mediated channel activity in odontoblast differentiation of hDPCs, and to explore the role and mechanism of Cx43 in regulating odontoblast differentiation of hDPCs induced by LPS. Statistical analysis was performed with SPSS 26.0 software package.

Results: IF results showed that Cx43 was mainly expressed in the odontoblast layer in healthy dental pulp tissues. At 3-24 h after tooth injury, the expression of Cx43 decreased and then gradually increased to the normal level; from 3 days to 2 weeks after injury, the expression of Cx43 tended to be down-regulated which was in the odontoblast layer and pulp proper. The expression of DSPP mRNA was significantly up-regulated in the hDPCs stimulated with 10 ng/mL LPS for 6 h(P＜0.01). Inhibition of Cx43 significantly up-regulated the expression of DSPP, DMP-1 and Osx mRNA induced by LPS in hDPCs(P＜0.05), while overexpression of Cx43 obviously inhibited the expression of factors related to LPS-induced odontoblast differentiation(P＜0.01) and the fluorescence intensity of DSPP. 10 ng/mL LPS activated ERK signal in hDPCs, and overexpression of Cx43 significantly attenuated the activity of ERK signal induced by LPS(P＜0.01). Inhibition of Cx43-mediated hemichannel (HC) promoted mRNA expression of factors related to odontoblast differentiation in hDPCs and the activity of ERK signal induced by LPS(P＜0.05), while blocking Cx43-mediated gap junction channel (GJC) inhibited odontoblast differentiation.

Conclusions: Cx43 participates in the regulation of dental pulp repair after injury, and its expression shows a downward trend as a whole. Inhibition of Cx43 or blocking of HC promotes LPS-induced ERK signal activity and odontoblast differentiation of hDPCs.

Purpose: To study the effect of different cleaning methods on the shear bond strength of self-adhesive resin cement to saliva-contaminated high translucency zirconia and surface wettability.

Methods: Eighty zirconia specimens were randomly divided into 5 groups (n=16), i.e., control group(not contaminated), 75% ethanol group,cleaning paste group,airborne-particle abrasion group, and atmospheric pressure cold plasma group. The contact angles was measured, shear bond strength were examined, and fracture types were determined. SPSS 26.0 software package was used for statistical analysis of the data.

Results: The atmospheric pressure cold plasma group produced the lowest contact angle(P＜0.05). The shear bond strength of the airborne-particle abrasion group, the cleaning paste group and the atmospheric pressure cold plasma group respectively were similar to the control group without significant difference(P＞0.05), while those were significantly higher than 75% ethanol group(P＜0.05). The mixed fracture mode of the atmospheric pressure cold plasma group evidently increased.

Conclusions: Airborne-particle abrasion, cleaning paste and atmospheric pressure cold plasma overcome the effects of saliva contamination, producing the shear bond strength to zirconia similar to the control group. The atmospheric pressure cold plasma improves hydrophilicity of high translucency zirconia significantly.

Purpose: Through questionnaire survey, parents' cognition of children's bad oral habits and their related influencing factors were explored, in order to provide a reference for science popularization and education in future work.

Methods: With a self-designed questionnaire, 247 parents of children at first visit were surveyed on basic information and problems related to bad oral habits. Descriptive statistics were used for the counting data. Logistic regression analysis was used for the relevant factors affecting the parents' cognition of children's bad oral habits with SPSS 26.0 software package.

Results: Among 247 parents of preschool children, 17.4% of the parents took their children to the hospital for treatment because of bad oral habits. The prevalence of oral unhealthy habits was 44%. Parents' knowledge of bad oral habits was not high, less than half of the parents (46.6%) knew about bad oral habits, of which 82.6% of the parents thought that bad oral habits would affect the development of children's teeth, jaws, face and mental health, including facial contour (62.1%), dentition (34.7%), masticatory function (48.4%), and mental health (21.1%). 78.3% of the parents thought that bad oral habits needed to be corrected; 69.6% of the parents thought that they needed to go to the hospital for treatment, and 30.4% of the parents thought that it was ok as long as their children giving up bad oral habits. 61.7% of the parents would seek medical treatment in time when their children had bad oral habits. The ways for parents to obtain knowledge about bad oral habits were hospital education (61.5%). Parents with different characteristics had different cognition of bad oral habits. Logistic regression analysis showed that parents' education background was a risk factor affecting parents' cognition of bad oral habits(P=0.009).

Conclusions: Parents' awareness of bad oral habits is not high, and parents' educational background is a risk factor for parents' awareness of bad oral habits. It is necessary to improve parents' awareness of oral habits, strengthen health education of bad oral habits, especially for parents with special signs, and improve the attention to oral health care, to achieve early detection, early prevention, early treatment and timely treatment, so as to prevent the occurrence and development of malocclusion.

Purpose: To investigate the effect of endoscopy-aided non-incisional periodontal regeneration technique (NIT) in the treatment of alveolar bone angular resorption.

Methods: Thirteen patients with severe periodontitis(13 diseased teeth) were selected. All patients had alveolar bone angular resorption on adjacent surface. The patients received NIT treatment 6 weeks after periodontal primary therapy. The visualization of subgingival environment was acquired by the periodontal endoscopy. Following the removal of the subgingival plaque, calculus and intra-bony granulation tissue, bone grafting materials were placed into the intra-bony defects with the assistance of a delicate gingival protector. No flap was elevated and no sutures were applied. Probing depth (PD), gingival recession (GR), clinical attachment level (CAL), as well as radiographic parameters were evaluated at baseline and 2 years after treatment. SPSS 22.0 software package was used for data analysis.

Results: At 2-years follow-up, an average CAL gain of (3.65±2.10) mm (P＜0.001), PD reduction of (4.42±1.66) mm (P＜0.001), and minimal increase in GR of (0.38±0.87) mm (P=0.25) were observed. Alveolar bone was significantly improved at 2-years follow-up on radiographs (P＜0.001).

Conclusions: For angular resorption site of alveolar bone, NIT treatment can obtain good periodontal regeneration results without flap inversion.

Purpose: To elucidate the disparities and similarities in the composition and function of fibroblast subtypes between normal oral mucosa and cutaneous tissue, to establish a unified classification of fibroblast subtypes between these two tissue types, comprehend the differences and similarities in their functionalities, and provide a foundational basis for future applications in the fields of tissue repair and regeneration.

Methods: Four single-cell databases from both oral mucosa and cutaneous tissue were integrated and fibroblast subpopulations were extracted. Batch effects were eliminated using Harmony, and fibroblast subpopulations were subsequently classified via UMAP (Uniform Manifold Approximation and Projection) clustering. The functional analysis of these subpopulations was conducted through gene set enrichment results. Statistical analysis was performed with R 4.2.0 software package.

Results: Eight distinct functional fibroblast subpopulations were defined, and their functions were found to be associated with the composition of the extracellular matrix, immunity, and contraction. Statistical analysis revealed differences in the composition ratios of these subpopulations between oral mucosa and skin tissue.

Conclusions: To evaluate the role of fibroblasts in tissue homeostasis and wound healing accomplished by integrating and analyzing fibroblasts from normal skin and oral mucosal tissue from various sites, this study identifies the differences in fibroblast subpopulation composition and function between these two tissue types in healthy conditions, and provides an understanding of oral mucosa and skin homeostasis and cellular function at the transcriptomic level. The findings of this study may serve as a basis for future research in this area.

Purpose: To observe the long-term clinical effect of implants retained complete overdentures with Locator attachments.

Methods: A total of 48 patients with edentulous jaws treated with implants retained complete overdentures with Locator attachments were selected from the Outpatient Department of Peking University School and Hospital of Stomatology from 2016 to 2017. Among them, 21 patients underwent double-maxillary complete overdentures restoration and 27 patients underwent single-maxillary restoration. A total of 230 implants were implanted. The clinical observation indicators included the implant survival rate, peri-implant mucosal bleeding on probing(BOP), the change in the vertical height of alveolar bone absorption around the implants, overdenture base fracture rate, artificial tooth fall off and fracture rate and other complications. The change of the locator attachment retention force of the implant-supported overdentures was evaluated. SPSS 13.0 software package was used for data analysis.

Results: During the five-years clinical observation period, 5 implants fell off, 1 narrow dimeter implant in the anterior zone was broken, and 12 implants were lost to follow-up. The implant survival rate was 97.25%. One year after the restoration therapy finished, peri-implant mucosal bleeding on probing (BOP+) was detected in 48 (21.4%) implants. The average BI was 0.21±0.42, which was higher in the anterior zone than that in the posterior zone. The vertical alveolar bone absorption height around the implants was (0.21±0.35) mm, 2 implants-supported complete overdenture bases were broken. After 5 years of restoration, 163(76.89%) implants had peri-implant mucosal bleeding on probing(BOP+). The average BI was 1.00±0.70, and the vertical alveolar bone absorption height around the implants was (0.58±0.85) mm. There was no significant difference between males and females. There was no significant difference in the peri-implant mucosal bleeding index and the alveolar bone vertical absorption height between the anterior zone and the posterior zone(P＞0.05). The mean BI of peri-implant mucosa and the vertical absorption height of peri-implant alveolar bone were significantly different between the 1-year observation period and the 5-year observation period respectively(P＜0.01). There were 17(26.15%) cases with overdenture bases fracture, and the fracture rate of artificial teeth was 16.92%. Most of them occurred in the midline area of the anterior zone and the location of the overdenture base on the locator attachments. The average first replacement time of the locator attachment nylon retainer washer was 34.2±10.3 months.

Conclusions: Implants retained complete overdentures with Locator attachments are effective in long term clinical observation. Complications are mainly found in peri-implant mucosal bleeding on probing and vertical alveolar bone absorption, and tended