Porphyromonas gingivalis, as a major pathogen of periodontitis, could rapidly adhere to and invade host gingival epithelial cells (GECs) for the induction of infection. One ATP-binding cassette (ABC) transporter gene was found to be upregulated during this infection process, however, the molecular mechanisms remain unclear. In this study, we systemically investigated the messenger RNA level changes of all ABC transporter family genes in P. gingivalis while being internalized within GECs by real-time polymerase chain reaction. We identified that two ABC transporter genes, PG_RS04465 (PG1010) and PG_RS07320 (PG1665), were significantly increased in P. gingivalis after coculturing with GECs. Mutant strains with knockout (KO) of these two genes were generated by homogenous recombination. PG_RS04465 and PG_RS07320 KO mutants showed no change in the growth of bacteria per se. Knockdown of PG_RS07320, but not PG_RS04465, caused decreased endotoxin level in the bacteria. In contrast, both mutant strains showed decreased Arg- and Lys-gingipains activities, with significantly reduced adhesion and invasion capabilities. Secreted interleukin-1β (IL-1β) and IL-6 levels in GECs cocultured with PG_RS04465 or PG_RS07320 KO mutants were also decreased, whereas, only the cells cocultured with PG_RS07320 KO mutants showed significant decrease. In addition, virulence study using mouse revealed that both KO mutant strains infection caused less mouse death than wild-type strains, showing reduced virulence of two KO strains. These results indicated that ABC transporter genes PG_RS04465 and PG_RS07320 are positive regulators of the virulence of P. gingivalis.
Objective:To explore whether TRPV1 in CeA of mice was involved in tooth movement pain and anxiety.Methods:For-ty 8-week-old adult male C57BL/6J mice were randomly divided into blank group,0 cN,10 cN,30 cN and 50 cN groups according to the tooth movement force.Then the behavioral indexes were recorded at different time points.The pain level of mice was evaluated by face wiping test,the anxiety-like state was evaluated by open field test and elevated maze test.Immunofluorescence method was used to observe the distribution and expression of TRPV1 in CeA,western blot method was used to detect the change trend of TRPV1 protein expression in mouse CeA during orthodontic tooth movement.Results:All the force values of mice showed pain during the experimental tooth movement,in which the 30 cN group showed the highest level of face wiping on the 1st day.The mice with different force values also showed different degrees of anxiety-like behavior,and the anxiety-like indexes in the 30 cN group were significantly higher than those in the 0 cN group on the 28th day(P＜0.05).Immunofluorescence staining showed that the expression of TRPV1 in the 30 cN 1 d group was higher than that in the 0 cN 1 d group(P＜0.001).Western blot experiment showed that the expression of TRPV1 in CeA in 30 cN group was significantly higher than that in 0 cN 1 d group during experimental tooth movement,and reached the peak on the 1st and 28th day(P＜0.001).Conclusions:TRPV1 in CeA participated in pain and anxiety of experimental tooth movement.
牙髓干细胞(DPSCs)是一类牙源性间充质干细胞,在一定影响因素作用下可分化为成牙本质细胞,形成修复性牙本质,这对临床治疗牙髓病及修复具有指导意义.本文系统性回顾了DPSCs成牙分化的影响因素,包括生物因素、化学因素、物理因素、基因转染、生物材料等,探究影响DPSCs向成牙本质细胞分化的条件,为龋病、牙髓及根尖周病的修复治疗以及其他领域的应用提供理论基础.
Objective:To investigate the expression pattern of cyclin-dependent kinase regulatory subunit 2(CKS2)and its effects on cell biological behavior in patients with oral squamous cell carcinoma(OSCC).Methods:Bioinformatics analysis was conducted to explore the expression level of CKS2 mRNA in OSCC and normal mucosal tissues.We selected 91 OSCC and 20 healthy oral mucosa samples.Immunohistochemical(IHC)staining was used to detect the expression level of CKS2 in 91 specimens of OSCC patients and 20 normal oral epithelial mucosa,and to explore the predictive value of CKS2 expression in OSCC patients.Small interfering RNA(siRNA)was used to silence CKS2 of Cal27 cells and its knockdown efficiency was determined by western blot.The effects of CKS2 si-lencing on the proliferation,cycle and migration of Cal27 cells were detected by cell clone formation assay,cycle assay and wound healing assay.Results:Bioinformatics analysis and IHC results showed that the expression of CKS2 in OSCC tissues was significantly higher than that of normal oral mucosa(P＜0.05).The expression of CKS2 was correlated with tumor size,cervical lymph node metas-tasis and survival rate of patients(P＜0.05).CKS2 silencing inhibited the proliferation and migration of Cal27 cells and induced G2 phase arrest(P＜0.05).Conclusions:Our findings revealed that CKS2 was overexpressed in OSCC,its high expression relating to the poor prognosis of OSCC patients,and involving in the positive regulation of OSCC cell proliferation,migration and cycle.
为了更快地反映国内外口腔生物医学研究的新进展、新动向、新成果,更好地满足广大读者和作者的需求,在征得主办、主管单位同意并经江苏省新闻出版局审批,《口腔生物医学》自 2024 年起刊期由季刊变更为双月刊,页码由66 页/期调整为60 页/期. 特此公告!
机械力在组织发育以及多种生理病理过程中起重要作用,许多分子、信号通路以及离子通道参与其中.口腔是一个开放的环境,处于口腔中的牙齿会不可避免地受到多种机械力的刺激,而牙发育是牙齿生长过程中最重要的一环,在这一过程中同样存在着力学因素,它们在不同程度上影响着牙发育过程.本文对牙齿发育过程中机械刺激响应的相关分子进行综述,从而为找到一种通过机械刺激促进牙发育及萌出过程的方法提供可能.
Objective:To observe and analyze the clinical effect of single-retainer cantilever resin-bonded bridge in replacing single anterior tooth missing.Methods:Twenty single-retainer cantilever resin-bonded bridges were fabricated for single anterior tooth miss-ing,and they were evaluated 6,12,24 and 36 months after restoration using the clinical evaluation criteria set by the American Public Health Association(APHA).At the same time,the periodontal indexes of the abutment tooth and the adjacent teeth before and after restoration were recorded to analyze the influence of the restoration on periodontal condition of the abutment teeth,and the VAS of pa-tient satisfaction was quantified.Results:In 20 cases of single retainer cantilever resin-bonded bridge,no debonding or fracture oc-curred during 36 months of repair period.1 case was classified as grade B due to edge coloring and the rest were graded A 24 months after repair;36 months after the completion of the restoration,3 cases were classified as grade B due to edge coloring,and the rest were grade A.The patients'satisfaction with the restoration effect was 100%,and there was a significant difference in patients'satisfac-tion with the status of missing tooth area before and after restoration(P＜0.05).And there was no significant difference in the periodon-tal indexes of the abutment tooth before and after restoration(P＞0.05).Conclusions:For patients with single anterior tooth missing,using a single retainer cantilever resin-bonded bridge for restoration can obtain a relatively stable restoration results.
Objective:Preparation of Nb2C/3D printed silicon nitride(Si3N4)composite tissue engineering scaffolds and investiga-tion on biological characteristics in vitro of Nb2C MXene nanocoating.Methods:Si3N4 scaffold was prepared by 3D printing technolo-gy,and Nb2C nanosheet suspension was prepared by HCl-LiF hydrothermal in situ corrosion and stripping treatment,Nb2C biocoating was obtained by soaking the Si3N4 scaffold in 1.5 mg/mL Nb2C nanosolution;Nb2C/Si3N4 scaffold extracts were prepared and filtered,and then divided into four groups with MC3T3 cell lines,including blank control,positive control with 2%DMSO,Si3N4 and Nb2C/Si3N4 groups,and cell proliferation activity was measured by CCK-8.Results:Compared to blank group,the proliferation activity of MC3T3 cells was significantly increased for positive control group(P＜0.05);Compared to Si3N4 group,the proliferation activity of MC3T3 cells was significantly increased for Nb2C/Si3N4 scaffold group(P＜0.05).Conclusions:Nb2C/Si3N4 composite scaffold bio-lating can promote the proliferation of MC3T3 cells.It is a material with potential application for bone tissue engineering.
Objective:Prepare Lianbu sustained-release gel,to examine its basic physicochemical properties,degree of release and toxicological properties,and to evaluate its pharmacodynamic effects in the treatment of experimental periodontitis.Methods:The pre-scription process for the preparation of Lianbu sustained-release gel was optimized by alkaline phosphatase assay to obtain the optimal mass ratio of the extract of coptis and the extract of rhizome drynariae.Glycerol,glyceryl triacetate,hydroxyethyl cellulose and poly-acrylic acid resin were used as gel matrix to prepare Lianbu sustained-release gel with 2%,4%,and 6%drug-loading rate,and their morphology,pH,stability,and the release rate in vitro were examined to optimize the optimal amount of drug loading.Acute toxicity test in mice at 20 g/kg by gavage using the maximum tolerated dose method.The mucosa of the lower lip of SD rats was sutured into a blind pouch,and then the sustained-release gel was injected into the pouch and applied to their lower gingival mucosa for local mucosal irritation experiments.Long-term toxicology experiments were conducted in SD rats by continuous gavage for 90 days to examine the blood routine and histopathological indexes.An experimental periodontitis model was established in SD rats,and the three-dimensional morphology of the alveolar bone of rats was observed using micro-CT,and the CEJ-ABC was measured.Results:The optimum mass ra-tio for the preparation of coptis extract and rhizome drynariae extract was 1 ∶ 10.The 2%,4%,and 6%Lianbu sustained-release gel all had good injectability,were in the neutral pH range,and were well stabilized.6%Lianbu sustained-release gel had the best effect on release.No acute toxic side effects were seen in mice given the sustained-release gel.The mucous membrane contact test did not show any significant irritant reactions such as erythema and edema.Long-term toxicology experiments showed no difference in general activity status,behavioral activities,diet,urine and stool,weight gain,hematology of rats in the Lianbu sustained-release gel group compared with the control group,and no obvious pathological changes were seen in the pathological examination of organs,indicating that the drug had no obvious toxic side effects.After treating experimental periodontitis rats for 4 weeks,micro-CT imaging results showed the alveolar bone resorption was significantly improved in 6%gel group and CEJ-ABC was significantly less than that in blank gel group and control group(P＜0.05).Conclusions:The prepared 6%Lianbu sustained-release gel had excellent physicochemical prop-erties and slow-release performance,and had certain therapeutic effects on experimental periodontitis rats.
近年来,在口腔医学研究领域,针对抗菌、去污、牙齿美白、抗肿瘤等应用需求,研究人员利用高级氧化技术(AOPs)的方法和特点,研发新型的治疗策略和技术.本文就相关的研究进展和应用潜力进行综述,为进一步深入该技术在口腔医学领域中的研究及临床转化提供思路和参考.
Objective:To explore the changes and clinical significance of miR-146a and miR-155 levels in peripheral blood of pa-tients with severe periodontitis and coronary heart disease.Methods:Thirty-two patients with severe periodontitis and coronary heart disease admitted to the hospital were selected as group A,another fifty-four patients with severe periodontitis were selected as group B,another fourty-five healthy individuals were selected as the control group.Enzyme-linked immunosorbent assay was used to detect labo-ratory indicators,including serum tumor necrosis factor-α(TNF-α),C-reactive protein(CRP),interleukin-6(IL-6),interleukin-1 β(IL-1β),total cholesterol(TC),triglycerides(TG),low-density lipoprotein(LDL),and high-density lipoprotein(HDL).Real time fluorescence quantitative PCR assay was used to detect the levels of miR-146a and miR-155 in peripheral blood.The univariate a-nalysis of factors affecting the occurrence of coronary heart disease in patients with severe periodontitis,and logistic regression analysis of the influencing factors.The receiver operating curve(ROC)was used to analyze the value of miR-146a and miR-155 levels in pre-dicting the occurrence of coronary heart disease in patients with severe periodontitis.Results:The levels of miR-146a in group A were higher than those of the control group and group B respectively(P＜0.05).The miR-155 levels in group A were lower than those in the control group and group B respectively(P＜0.05).The body mass index,serum CRP,and miR-146a levels of patients in group A were higher than those in group B(P＜0.05),while the education years and miR-155 levels of patients in group A were lower than those in group B(P＜0.05).Logistic multivariate regression analysis showed that the average level of miR-146a was an independent risk factor for the occurrence of coronary heart disease in patients with severe periodontitis,but the level of miR-155 was a protective factor(P＜0.05).ROC analysis showed that the optimal cutoff points for miR-146a and miR-155 levels in peripheral blood predicting the oc-currence of coronary heart disease in patients with severe periodontitis were 1.84 and 0.60 respectively,the sensitivity was 81.25%and 84.37%respectively,and the specificity was 83.33%and 72.22%respectively.Conclusions:The miR-146a and miR-155 levels experience abnormal changes in patients with severe periodontitis and coronary heart disease,miR-146a and miR-155 levels can serve as sensitive indicators for predicting the occurrence of coronary heart disease in patients with severe periodontitis,and the combined predictive value is higher.
Objective:To identify potential diagnostic markers and assess their role in the pathogenesis and progress of temporoman-dibular joint(TMJ)pigmented villous nodular synovitis(PVNS)based on bioinformatics analysis.Methods:The gene expression pro-file was analyzed at the probe level according to the raw matrix data of gene chip(GSE3698).R language programming was used to an-alyze and visualize the results.DEGs were screened in TMJ PVNS.The functional annotation and signal pathway enrichment analysis on these cell populations.PPI network was established based on the STRING database.Summary receiver operating characteristic curves were plotted to determine diagnostic biomarkers of TMJ PVNS.Quantitative analysis of immune cell subtypes in TMJ PVNS was calcu-lated using CIBERSORT software from sequences and the association between diagnostic markers and infiltrating immune cells was also estimated accordingly.Results:Totally 139 DEGs were found out including 46 downregulated and 93 upregulated genes.Functional en-richment analysis revealed that 44 biological process,30 cellular component,18 molecular function,and 22 signal pathways were sta-tistically significant.Eight hub genes with significantly upregulated expression were screened by PPI network.FCER1G,HLA-DPB1,LAPTM5,and TYROBP were identified as potential diagnostic biomarkers for TMJ PVNS.Immunocyte infiltration analysis showed neutrophils and M2 macrophages were associated with the pathogenesis and progress of TMJ PVNS.In addition,the correlation analysis con-cerning the identified diagnostic markers and infiltrating immune cell subpopulations indicated that FCER1G,LAPTM5,TYROBP were negatively related to plasma cell infiltration;FCER1G,HLA-DPB1,LAPTM5,and TYROBP were positively related to M2 macrophage infiltration and that neutrophils.Conclusions:This investigation concluded that FCER1G,HLA-DPB1,LAPTM5,and TYROBP as potential diagnostic markers and immune cell infiltration played an important role in the pathogenesis and progress of TMJ PVNS.
目的:探究骨髓巨噬细胞主穹窿蛋白(MVP)对骨再生修复的影响.方法:在野生型(WT)C57BL/6 小鼠和特异性敲除巨噬细胞MVP(CKO)的小鼠中分别构建胫骨骨皮质缺损模型,通过Micro-CT扫描与重建技术、苏木素-伊红染色观察骨再生情况;体外分离培养WT和CKO小鼠骨髓来源的巨噬细胞,并分别与野生型骨髓间充质干细胞(BMSC)共培养,通过碱性磷酸酶染色、成骨分化标记基因实时荧光定量PCR实验,观察敲除巨噬细胞MVP对共培养体系中的BMSC成骨向分化的调控作用;通过细胞因子芯片实验检测巨噬细胞细胞因子的分泌.结果:骨缺损第 14 天,Micro-CT扫描及重建结果显示CKO小鼠缺损区矿化组织少于WT小鼠(P＜0.05);苏木素-伊红染色结果显示CKO组小鼠缺损区骨痂面积小于WT小鼠;与CKO组巨噬细胞共培养的BMSC的成骨分化标记基因水平显著低于与WT组巨噬细胞共培养组(P＜0.05);敲除MVP的巨噬细胞分泌更多抑制成骨的细胞因子CCL2、CCL4、CXCL10、CCL3、IL-7.结论:巨噬细胞MVP可能通过促进BMSC成骨分化进而调控骨再生修复.
修复再生受损的牙髓组织,恢复患牙天然的牙髓功能,逐渐成为治疗牙髓疾病的新的重要目标.浓缩生长因子是第3 代经自体全血离心后获得的血小板浓缩制品,富含大量三维网状结构的纤维蛋白和丰富的生长因子,对组织再生具有很强的促进作用,可作为牙髓再生的良好支架材料.本文就浓缩生长因子在牙髓组织再生中的作用机制进行综述,旨在为进一步的研究和临床应用提供理论支持.
目的:通过磁共振成像(MRI)测量cN0 期患者舌部病灶的浸润深度及肿瘤体积,探索其在患者颈淋巴结转移及预后中的预测价值.方法:收集术前完善口咽MRI检查的cN0 期舌鳞癌患者74 例,通过IKT-SNAP和3D slicer软件重建舌部病灶并测量其浸润深度及肿瘤体积;通过R语言绘制浸润深度、肿瘤体积与颈淋巴转移之间的受试者工作特征曲线(ROC)曲线并确定临界值;对临床参数与颈淋巴转移的相关性进行Logistic回归分析,并通过单因素及多因素COX回归分析影响cN0 期舌鳞癌患者预后的独立危险因素.结果:6.3 cm3的肿瘤体积和 14 mm的肿瘤浸润深度可作为临界值来预测颈淋巴结的转移;多因素回归分析表明MRI测量得到的浸润深度可作为影响舌鳞癌患者颈淋巴结转移和预后的独立危险因素(P＜0.05);Kaplan-Meier分析表明浸润深度≥14 mm的患者有着更差的预后(P＜0.05).结论:MRI测得的浸润深度可作为预测cN0 期舌鳞癌患者颈淋巴结转移和预后的独立指标,而肿瘤体积则没有这样的预测作用.
目的:探讨雌激素缺乏对大鼠根尖牙乳头干细胞(SCAPs)增殖及牙向/骨向分化能力的影响.方法:选取 20 只 8 周龄雌性SD大鼠随机分为假手术(Sham)组和卵巢去势(OVX)组,每组各 10 只.全麻下OVX组摘除双侧卵巢,Sham组行相同切口保留双侧卵巢,术前、术后 1 个月分别检测两组大鼠血清雌二醇水平.1 个月后每组各处死 5 只大鼠,分离切牙根尖牙乳头组织,酶消化法分离培养两组SCAPs,采用MTT、碱性磷酸酶(ALP)活性检测、实时荧光定量PCR、Western blot检测雌激素缺乏对大鼠SCAPs增殖及成牙/成骨向分化能力的影响.将两组SCAPs细胞以明胶海绵为载体分别植入对应组别大鼠肾被膜下,8 周后取出植入组织,使用数字化X线牙片机对取出团块曝光成像及HE染色观察其矿化能力.结果:OVX组大鼠去势1 个月后血清雌二醇水平显著低于术前(P＜0.001),Sham组大鼠血清雌二醇水平术前、术后差异无统计学意义(P＞0.05).MTT结果显示,OVX组SCAPs增殖能力显著下降(P＜0.001).ALP活性检测结果显示,OVX组SCAPs的ALP活性低于Sham组(P＜0.001).实时荧光定量PCR和Western blot结果均表明OVX组SCAPs的牙本质涎磷蛋白(DSPP)、成骨细胞特异性转录因子Osterix(OSX)、骨钙素(OCN)等成牙/成骨相关基因和蛋白表达低于Sham组(P＜0.01).体内移植结果显示,Sham组形成密度较高的较规则牙髓-牙本质复合体,而OVX组形成不规则结构.结论:雌激素缺乏减弱大鼠SCAPs的增殖及牙向/骨向分化能力.
目的:探索结缔组织生长因子(CTGF)对尼克样 1 型蛋白(NELL-1)诱导的MC3T3-E1 细胞成骨分化能力的影响.方法:以MC3T3-E1 细胞为研究对象,分别使用PBS(对照)、NELL-1、CTGF和NELL-1+CTGF处理细胞,利用CCK-8 法检测各组细胞增殖情况;成骨诱导后,利用茜素红染色和定量分析、碱性磷酸酶(ALP)活性检测和ALP 染色、实时荧光定量PCR和Western blot实验检测各组细胞成骨分化情况.结果:与对照组相比,CTGF一定程度上抑制MC3T3-E1 细胞增殖;NELL-1单独处理可诱导细胞形成更多矿化结节,增强ALP活性,加深ALP染色,明显提高Runt相关转录因子 2(RUNX2)、ALP、成骨细胞特异性转录因子Osterix(OSX)等成骨形成基因的转录和蛋白表达水平(P＜0.01).与NELL-1单独处理组相比,NELL-1+CTGF联合处理组中细胞内矿化结节形成减少,ALP活性降低,ALP染色变浅,RUNX2、ALP、OSX等基因的转录和蛋白表达水平显著降低(P＜0.05).结论:CTGF抑制NELL-1诱导MC3T3-E1 细胞的成骨分化能力.
目的:应用三维有限元研究翼上颌种植设计在上颌后牙区牙列缺损种植修复中骨组织及种植体应力分布的变化.方法:在六组上颌后牙区牙列缺损的颌骨模型中,第二前磨牙位置植入一枚常规植体,避开上颌窦在翼上颌区域分别以咬合平面 45°或以Frankfort平面 70°倾斜植入一枚种植体,制作三单位种植桥修复体.以 200 N的静态力垂直加载于后牙功能尖上,应用软件分析翼上颌种植体及周围骨组织的应力.结果:70°倾斜种植后,颌骨应力为(43.18±17.15)MPa;45°倾斜种植时,颌骨应力为(67.25±18.44)MPa.70°倾斜种植配合近中种植体形成固定桥联冠修复体,其跨度大于三单位,在第二磨牙加载垂直向力 200 N后,颌骨应力为(32.49±19.01)MPa,最大应力位于倾斜植入的种植体穿出位点处;在第一磨牙加载垂直向力200 N后,颌骨应力为(26.89±14.66)MPa,最大应力位于近中种植体穿出位点处.45°倾斜种植配合近中种植体形成固定桥联冠修复体,其跨度约为三单位,在第二磨牙的颌骨应力为(39.88±17.12)MPa,最大应力位于倾斜植入的种植体穿出位点处;在第一磨牙的颌骨应力为(31.03±15.05)MPa,最大应力位于近中种植体穿出位点处.结论:翼上颌区种植体可 70°倾斜植入时,上部修复体可选择单冠修复或固定桥联冠修复;种植体可 45°倾斜植入时,上部修复体须选择固定桥联冠修复.
