Osteoblasts and osteoclasts collaborate in bone metabolism, facilitating bone development, maintaining normal bone density and strength, and aiding in the repair of pathological damage. Endoplasmic reticulum stress (ERS) can disrupt the intracellular equilibrium between osteoclast and osteoblast, resulting in dysfunctional bone metabolism. The inositol-requiring enzyme-1α (IRE1α) pathway-the most conservative unfolded protein response pathway activated by ERS-is crucial in regulating cell metabolism. This involvement encompasses functions such as inflammation, autophagy, and apoptosis. Many studies have highlighted the potential roles of the IRE1α pathway in osteoblasts, chondrocytes, and osteoclasts and its implication in certain bone-related diseases. These findings suggest that it may serve as a mediator for bone metabolism. However, relevant reviews on the role of the IRE1α pathway in bone metabolism remain unavailable. Therefore, this review aims to explore recent research that elucidated the intricate roles of the IRE1α pathway in bone metabolism, specifically in osteogenesis, chondrogenesis, osteoclastogenesis, and osteo-immunology. The findings may provide novel insights into regulating bone metabolism and treating bone-related diseases.

OBJECTIVES:This study aimed to report the clinical and radiographic results of 2.8 mm two-piece narrow diameter implant (NDI) supporting fixed restorations. MATERIALS AND METHODS:Clinical and radiographic data of 54 NDIs in 32 patients were retrospectively assessed after 2 to 11 (mean 8.17) years of follow-up. Clinical and radiographic measurements were taken. Survival rate, implant and prosthesis failure, pink aesthetic scores (PES), white aesthetic scores (WES), bleeding on probing (BOP), probing depth (PD), marginal bone loss (MBL), and mechanical and biological complications were evaluated. RESULTS:An implant failed during the follow-up period, resulting in a cumulative survival rate of 98.15% at the implant level and 96.88% in the patient. The total mean values of PES and WES for 2.8 mm NDIs were 7.09 ± 1.15 (range: 3.33-9.00) and 7.42 ± 1.03 (range: 3.67-9.33). The prevalence of sites with positive BOP was 38.14 ± 29.77%. The mean PD value was 2.46 ± 0.62 mm. The average MBL was 1.15 ± 0.74 mm (range: 0.25-4.03 mm). No implant or abutment fracture was detected. A veneer chipping was present in one patient, and a loose crown appeared in another patient. Two implants (3.7%) and two patients (6.3%) were diagnosed with peri-implantitis. CONCLUSION:Within the limitation of the study, the results indicate that the use of two-piece 2.8 mm NDI for the fixed prosthetic rehabilitation of edentulous regions with reduced interdental and/or buccal-lingual width is viable.

Abstract Background Arteriovenous fistula (AVF) maturation is a process involving remodeling of venous arm of the AVFs. It is a challenge to balance adaptive AVF remodeling and neointima formation. In this study we temporally controlled Notch activation to promote AVF maturation while avoiding neointima formation. Methods Temporal Notch activation was controlled by regulating the expression of Notch transcription factor, RBP-Jκ, or dnMAML1 (dominant negative MAML2) in vascular smooth muscle cells (VSMCs). AVF mouse model was created and VSMC phenotype dynamic changes during AVF remodeling were determined. Results Activated Notch was found in the nuclei of neointimal VSMCs in AVFs from uremic mice. We found that the VSMCs near the anastomosis became dedifferentiated and activated after AVF creation. These dedifferentiated VSMCs regained smooth muscle contractile markers later during AVF remodeling. However, global or VSMC-specific KO of RBP-Jκ at early stage (before or 1 week after AVF surgery) blocked VSMC differentiation and neointima formation in AVFs. These un-matured AVFs showed less intact endothelium and increased infiltration of inflammatory cells. Consequently, the VSMC fate in the neointima was completely shut down, leading to an un-arterialized AVF. In contrast, KO of RBP-Jκ at late stage (3 weeks after AVF surgery), it could not block neointima formation and vascular stenosis. Inhibition of Notch activation at week 1 or 2, could maintain VSMC contractile markers expression and facilitate AVF maturation. Conclusions This work uncovers the molecular and cellular events in each segment of AVF remodeling and found that neither sustained increasing nor blocking of Notch signaling improves AVF maturation. It highlights a novel strategy to improve AVF patency: temporally controlled Notch activation can achieve a balance between adaptive AVF remodeling and neointima formation to improve AVF maturation. Translational perspective Adaptive vascular remodeling is required for AVF maturation. The balance of wall thickening of the vein and neointima formation in AVF determines the fate of AVF function. Sustained activation of Notch signaling in VSMCs promotes neointima formation, while deficiency of Notch signaling at early stage during AVF remodeling prevents VSMC accumulation and differentiation from forming a functional AVFs. These responses also delay EC regeneration and impair EC barrier function with increased inflammation leading to failed vascular remodeling of AVFs. Thus, a strategy to temporal regulate Notch activation will improve AVF maturation.

目的 研究细丝蛋白B(FLNB)在钙化性心脏瓣膜中的表达及其临床意义.方法 2021年8月～2022年1月在我院手术切除的钙化性主动脉瓣标本9例,同期因主动脉夹层行手术切除的正常主动脉瓣膜标本8例作为对照.组织切片茜素红染色观察瓣膜钙化结节.采用免疫组化分析方法对瓣膜中细丝蛋白B的表达进行分析.利用冰冻组织标本进行蛋白质免疫印迹检测,验证相应分子表达水平间差异.结果 钙化瓣膜切片经茜素红染色可以看见明显钙化结节.免疫组化检查显示细丝蛋白B表达于细胞质,钙化瓣膜高于正常瓣膜,钙化组平均光密度为(6.57±1.65)％;对照组为(0.91±0.52)％,两组比较差异有统计学意义(P<0.05),蛋白质免疫印迹结果与之一致.结论 细丝蛋白B在钙化瓣膜中表达增高,可能参与了瓣膜钙化的病理进程.

目的:探讨医患沟通教育在口腔修复学教学中的应用效果.方法:将口腔修复科实习的五年制口腔医学本科生随机分为2组:实验教学组(n=25)采用医患沟通教育融合常规教学模式;传统教学组(n=25)采用常规教学模式.利用医患沟通行为量表测评,教学效果调查问卷比较两种教学模式的效果.结果:实验教学组医患沟通能力优于传统教学组(P<0.05),医学生也更喜欢融合教育模式(100％).结论:将医患沟通教育融入口腔修复传统教学中有利于提高医学生医患沟通能力和教学满意度.

目的:检测并分析人脐带间充质干细胞(hUCMSCs)成骨分化过程中的成骨特征性miRNA表达谱,探讨其可能的靶基因和临床意义.方法:对Shh诱导hUCMSCs成骨分化过程中表达的miRNA进行高通量测序分析,检测其表达量的变化,并验证成骨相关miRNA的表达差异.结果:通过miRNA高通量测序和RT-qPCR验证,预测miRNA-342-3p可能作为关键因子,通过靶向Sufu调控Shh信号通路,进而调控hUCMSCs的成骨分化.结论:筛选出新的调控因子miRNA-342-3p作为Shh信号通路的靶分子,对成骨分化过程进行精准调控,为种植义齿骨组织工程提供新的治疗思路.

目的:探究大气压常温等离子体(atmospheric room temperature plasma,ARTP)对人牙周膜干细胞(hu-man periodontal ligament stem cells,hPDLSCs)成骨分化的影响.方法:原代培养人牙周膜细胞,采用免疫磁珠法从中分选出hPDLSCs,通过成骨和成脂诱导培养检测其分化潜能;利用不同时长ARTP处理hPDLSCs,行成骨诱导培养,测定细胞碱性磷酸酶(alkaline phosphatase,ALP)活性,茜素红染色(alizarin red staining,ARS)及半定量分析法观察细胞矿化结节形成状况,RT-qPCR测定细胞成骨相关基因的表达状况,测定细胞内活性氧粒子(reactive oxygen species,ROS)含量变化.结果:采用免疫磁珠法分选出的hPDLSCs呈长梭形、多角形,表现出良好的成骨和成脂分化潜能;ARTP处理时长1 min可提高hPDLSCs的ALP活性,增加细胞内矿化结节形成量;能够提升hPDLSCs成骨相关基因ALP、COL-I、RUNX2的表达水平;ARTP提高了hPDLSCs胞内ROS水平.结论:特定处理时长的ARTP能够促进hPDLSCs成骨分化,ROS可能在该过程发挥作用.ARTP有望成为颌面骨再生治疗的潜在方法之一.

目的:观察与支架材料三维培养后,可控式碱性成纤维细胞生长因子(bFGF)联合音猬因子(Shh)的时序基因传递对骨髓间充质干细胞促成骨效应的调控作用.方法:使用受四环素调控的重组腺相关病毒rAAV2-tet-off-bFGF以及rAAV2-Shh共同感染大鼠骨髓间充质干细胞(BMSCs),体外与β-磷酸三钙(β-TCP)复合三维培养.扫描电镜观察细胞附着于材料表面的形貌.MTT检测复合物中细胞的增殖状况.实时定量PCR检测复合物中细胞内各成骨相关因子mRNA的表达变化.结果:扫描电镜可见三维培养后15 h两组细胞已黏附于材料表面,至培养后4 d,细胞呈多角形向材料内部孔隙伸展.MTT显示转基因BMSCs与β-TCP复合三维培养5 d后细胞增殖明显,与其它两组比较均有统计学差异.实时定量PCR及碱性磷酸酶检测发现双因子时序表达组,不管贴壁培养还是三维培养,胞内成骨相关因子的表达与对照组相比均有显著升高.结论:四环素调控下能成功实现bFGF及Shh在BMSCs中的时序表达,与β-TCP复合三维培养后,转基因细胞的增殖速度、成骨相关因子的表达均有显著提升.

Aims It is well-established that endothelial dysfunction promotes activation of vascular smooth muscle cell (VSMC). Whether decreased accumulation of VSMCs affects endothelial regeneration and functions in arteriovenous graft (AVG) remodelling has not been studied. We sought to identify mechanisms by which the Notch ligand, Jagged1, in VSMCs regulates endothelial cell (EC) functions in AVGs. Methods and results AVGs were created in transgenic mice bearing VSMC-specific knockout (KO) or overexpression of Jagged1. VSMC migration, EC regeneration, and its barrier functions as well as AVG remodelling were evaluated. Jagged1 expression was induced in VSMCs of neointima in the AVGs. Jagged1 KO in VSMCs inhibited the accumulation of extracellular matrix as well as VSMC migration. Fewer α-SMA-positive VSMCs were found in AVGs created in VSMC-specific Jagged1 KO mice (VSMCJagged1 KO mice) vs. in WT mice. Decreased VSMCs in AVGs were associated with deterioration of EC functions. In AVGs created in transgenic mice bearing Jagged1 KO in VSMCs exhibited delayed EC regeneration and impaired EC barrier function. Barrier dysfunction of ECs increased inflammatory cell infiltration and dysregulation of AVG remodelling and arterialization. The increased expression of IL-1β in macrophages was associated with expression of adhesion markers in ECs in AVGs created in VSMCJagged1 KO mice. In contrast, AVGs created in mice with overexpression of Jagged1 in VSMCs exhibited improved EC regeneration plus decreased macrophage infiltration. This led to AVG remodelling and arterialization. In co-cultures of ECs and VSMCs, Jagged1 deficiency in VSMCs suppressed N-cadherin and integrin β3 expression in ECs. Inhibition of integrin β3 activation delayed EC spreading and migration. Notably, Jagged1 overexpression in VSMCs or treatment with recombinant Jagged1 stimulated the expression of N-cadherin and integrin β3 in ECs. Jagged1-induced responses were blocked by inhibition of Notch signalling. Conclusions Jagged1 expression in VSMCs maintains EC barrier functions and blocks infiltration of macrophages. These responses promote remodelling and arterialization of AVGs.

Chronic kidney disease (CKD) induces the failure of arteriovenous fistulas (AVFs) and promotes the differentiation of vascular adventitial GLI1-positive mesenchymal stem cells (GMCs). However, the roles of GMCs in forming neointima in AVFs remain unknown. GMCs isolated from CKD mice showed increased potential capacity of differentiation into myofibroblast-like cells. Increased activation of expression of PDGFRA and hedgehog (HH) signaling were detected in adventitial cells of AVFs from patients with end-stage kidney disease and CKD mice. PDGFRA was translocated and accumulated in early endosome when sonic hedgehog was overexpressed. In endosome, PDGFRA-mediated activation of TGFB1/SMAD signaling promoted the differentiation of GMCs into myofibroblasts, extracellular matrix deposition, and vascular fibrosis. These responses resulted in neointima formation and AVF failure. KO of Pdgfra or inhibition of HH signaling in GMCs suppressed the differentiation of GMCs into myofibroblasts. In vivo, specific KO of Pdgfra inhibited GMC activation and vascular fibrosis, resulting in suppression of neointima formation and improvement of AVF patency despite CKD. Our findings could yield strategies for maintaining AVF functions.

OBJECTIVE:Previously, we have shown that miRNA-342-3p was increased during osteogenic differentiation of human umbilical mesenchymal stem cells (hUCMSCs) via regulating the sonic hedgehog (Shh) pathway. In this study, our objective is to further investigate the role of miRNA-342-3p in activation of Shh pathway by targeting suppressor of fused protein (Sufu), a suppressor of transcriptional factor Gli, as well as the potential interaction with transforming growth factor beta (TGF-β) signaling pathway during osteogenic induction of hUCMSCs. MATERIALS AND METHODS:HUCMSCs that stable overexpression or knockdown of miRNA-342-3p were established by infection with lentiviral vectors. mRNA and protein levels of Hedgehog signaling pathway and osteogenic genes were measured by RT-qPCR and western blot assays. Luciferase reporter assay was performed to test the direct binding site of Sufu 5'UTR targeted by miRNA-342-3p. RESULTS:Overexpression of miRNA-342-3p in hUCMSCs enhanced the expression of osteogenic genes by targeting Sufu. And the potential of osteogenic differentiation of hUCMSCs was inhibited while knocking down miRNA-342-3p. Meanwhile, induced the TGF-β expression level was also observed upon overexpressing miRNA-342-3p, suggesting activation of TGF-β signaling pathway was a potential mechanism of miRNA-342-3p-mediated osteogenesis in hUCMSCs. CONCLUSIONS:Our findings provide new mechanistic evidence that miRNA-342-3p might be a valuable therapeutic target in bone regeneration.

Human umbilical mesenchymal stem cells (UCMSCs) have been wildly used in tissue engineering field as a promising source because of their unlimited and noninvasive origin. To selectively induce osteogenic differentiation of UCMSCs, it's imperative to understand the regulatory molecular mechanism underlying the process of how these cells switch into osteogenic lineage path. We previously showed enhanced sonic hedgehog (Shh) signaling pathway upon osteogenic induction in mesenchymal stem cells. In this study, miRNA-seq analysis revealed substantial Shh-dependent expression of distinct miRNAs, including miR-342-3p, during ostogenesis. RT-qPCR confirmed that miR-342-3p was increased at a greater level when Shh signaling pathway was activated by N-terminal of Shh ligand compared with osteogenic induction alone, in contrast to the decreasing of suppressor-of-fused protein (Sufu). Consistently, transient overexpressing miR342-3p in UCMSCs via miR-342-3p mimics dramatically decreased Sufu, a suppressor of Gli, while osteogenic markers, including alkaline phosphate and osteocalcin, were upregulated during osteogenic induction, indicating that miR-342-3p might be involved in osteogenesis through the Shh signaling pathway. In conclusion, this study showed the potential of miR-342-3p as a therapeutic target to promote bone regeneration by modulating expression of Sufu in UCMSCs.

Objective:The aim of this study was to isolate and identify the human umbilical cord mesenchymal stem cells (hUCMSCs). Method:Umbilical cord was cut into pieces, and then digested with collagenaseII. The mesenchymal stem cells from human umbilical cord were cultured. After that, the multipotent differentiation characteristics were demonstrated using standard in vitro conditions. Furthermore,the biological properties of hUCMSCs were investigated by flow cytometry. And the relevant mRNAs levels of Hedgehog signaling pathway were detected by real-time PCR. Result:Monoptychial het-erogeneous cells were obtained.The cells were able to differentiate into osteogenic, adipogenic and cartilage cells. A total of 98.4 % of cells at passage 3 expressed cluster of differentiation CD29 and 98.80% of CD44,85.09% of CD105, but only 0.19%of CD45 and 0.58%of CD34.And expression of the factors of hedgehog signaling pathway were able to be detected by real-time PCR. Conclusion:hUCMSCs that are isolated from the human umbilical cord and exhibit the identified character-istics may be used as seed cells in jaw bone defect reconstruction.

The aim of this study was to investigate the cytotoxicity of modified nonequilibrium plasma with chlorhexidine digluconate (CHX) on human gingival fibroblasts (HGFs), and to evaluate the biosecurity of modified nonequilibrium plasma with 2% CHX as a new method of root canal treatment. Tissue samples taken from human gingiva were primarily cultured and passaged. Cells from passages 3–7 were used. HGFs were treated by modified nonequilibrium plasma with 2% CHX for 0 min (control group), 30 s, 1 min, 1.5 min, 3 min, 5 min, and 10 min, respectively, and then they were incubated for 0, 24, and 48 h. After that, cell counting kit-8 (CCK-8) assay was applied to analyze the cytotoxicity of modified nonequilibrium plasma with 2% CHX on HGFs. There was no significant difference between the 0 h group treated with the modified nonequilibrium plasma for 1 min and the control group (P>0.05). However, there were significant differences between all the other treated groups and the control group (P<0.05). When treated for 1.5 min or shorter, the cell viability was obviously increased; while treated for 3 min or longer, it was obviously reduced. Moreover, when successively cultured for 0, 24, and 48 h, cell viability was decreased at first and then increased in the 3-min-treated and 5-min-treated groups. The modified nonequilibrium plasma with 2% CHX was of no influence on cell viability in 1.5 min treatment, and it could be safely used on root canal treatment.

目的:制备携带碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)的病毒载体,能被四环素调控表达.方法:构建重组质粒pAAV-S3-bFGF,生产重组病毒rAAV2-tet-off-bFGF,并感染小鼠颅骨前成骨细胞MC3T3-E1,实时定量PCR检测强力霉素(doxycycline,Dox)调控下rAAV2-tet-off-bFGF对MC3T3-E1中成骨相关因子的影响.结果:生产出高滴度及纯度的重组病毒rAAV2-tet-off-bFGF,能被Dox调控,感染MC3T3-E1后能促进胞内成骨相关因子的表达升高.结论:这种重组病毒rAAV2-tet-off-bFGF能在Dox调控下有效传递外源基因,为骨组织工程中的种子细胞提供基因传递载体.

Objective:The aim of this study was to delivery basic fibroblast growth factor(bFGF) and sonic hedgehog (Shh) to bone marrow derived mesenchymal stem cells (BMSCs),and then to detect the effect for osteogenic marker genes. Method:bFGF and Shh were constructed to the recombinant adeno-associated virus,respectively(rAAV2-tet-off-bFGF and rAAV2-Shh). The previous viral vector allowed for regulation of the bFGF expression by the addition of doxycycline,a te-tracycline analogue. Several osteogenic markers were detected by quantitative real-time reverse transcriptase polymerase chain reaction and western blot. Result:BMSCs were constructed successfully by two recombinant virus that were regulated by Dox. The expression of osteogenic marker mRNA and protein had an increased tendency after two genes transduction(P<0.05). Conclusion:Sequential delivery of angiogenic and osteogenic factors likely has a synergistic effect,which could not only enhance the cell proliferation,but also improve the expression of osteogenic marker of BMSCs.

Objective:The aim of this study was to cultivate and determine the bone marrow derived mesenchymal stem cells (BMSCs). Method:Bone marrow harvested from rat femurs were cultured. BMSCs were selected and expanded through passaging in vitro. Furthermore,the biological properties of BMSCs were investigated by flow cytometry. Meanwhile the multipotent differentiation characteristics were demonstrated using standard in vitro conditions. Result:Monoptychial heterogeneous cells were obtained. A total of 98.4 % of cells at passage 3 expressed cluster of differentiation (CD)29 and CD90,but not CD45. The cells were able to differentiate into osteogenic and adipogenic cells. Conclusion:BMSCs that are isolated from the rat bone marrow and exhibit the identified characteristics may be used as seed cells in bone tissue engineering.