OBJECTIVE:Complement component 5a (C5a) binding to its receptor (C5aR) can mediate pro-inflammatory immune responses, which impairs osteogenic differentiation of dental pulp stem cells (DPSCs). The present study aimed to evaluate the regulatory effect of C5aR antagonist (W54011) on the osteogenic differentiation of DPSCs in an in vitro inflammatory microenvironment and its reparative effect on in vivo LPS-induced inflammatory mandibular defects, to provide novel strategies for bone regeneration under inflammatory conditions. MATERIALS AND METHODS:In vitro, a DPSCs inflammatory model was established with 1.0 μg/mL LPS for 48 h, and cells were divided into four groups (LPS + W54011 + OIM, OIM, W54011 only, blank) with W54011 at 1.0 μg/mL. Each assay included 3 biological and 3 technical replicates, and osteogenic differentiation was assessed via Alkaline Phosphatase (ALP) staining, alizarin red S (ARS) staining, RT-qPCR, and western blotting for OCN, Osterix, and RUNX2. In vivo, an LPS-induced inflammatory mandibular defect model was constructed in Sprague-Dawley (SD) rats; after 2 weeks of inflammation, defects were implanted with W54011-loaded Bio-Oss bone powder, and healing was evaluated by micro-CT, H-E staining, and immunohistochemistry for IL-6, OCN, and RUNX2. RESULTS:In vitro, all 4 groups showed positive ALP and mineralized nodule staining. Molecular biology methods found differential OCN, Osterix and RUNX2 expression at day 7, 14 and 21, with the LPS + W54011 + OIM group having higher osteogenesis markers at certain times (p < 0.05, one-way ANOVA with Bonferroni post-hoc test). In vivo, Micro-CT and H&E staining revealed that the defect site implanted with W54011-loaded Bio-Oss (treatment group) exhibited higher bone density and denser new bone formation compared to the control groups. However, due to the small sample size, these results were considered preliminary qualitative observations without statistical analysis. CONCLUSION:1. LPS (1 μg/ml) stimulation for 48 h can be used to establish a DPSCs inflammation model. 2. W54011 can promote the osteogenic differentiation of DPSCs in an inflammatory environment in vitro. 3. Preliminary in vivo data suggest that W54011-loaded Bio-Oss may facilitate the healing of inflammatory mandibular defects, warranting further investigation with larger samples.

Deep tooth decay approaching the pulp may develop into pulpitis; to prevent this, pulp cells need to balance the rapid immune response to avoid rapid swelling of the pulp. Current treatment of deep decay that approaches the pulp involves the application of drugs that induce low‐level inflammation in the dental pulp to promote its repair, but this treatment is sometimes insufficient. However, the unsuccessful treatment often resulted in pulpitis. The C5a‐C5aR is the initial stage of the immune cascade response. Blocking the binding of C5a‐C5aR can slow the immune response in the narrow pulp cavity, so that dental pulp cells have enough time to proliferate, migrate, and differentiate. In this study, we compared lipoteichoic acid (LTA) and lipopolysaccharides (LPS) at different concentrations and time points and used the C5aR antagonist W54011 to block the C5a‐C5aR axis. The blocking effect was detected by analyzing the expression of C5a, C5aR, interleukin (IL)‐6, and Toll‐like receptors 2 and 4 (TLR‐2, 4). Next, we determined the optimal concentration and duration of LTA and LPS treatment in combination with W54011. Based on our results, we selected 1.0 μg·mL−1 LPS treatment for 48 h to generate an inflammatory model of human dental pulp cells. We then regrouped the cells and conducted expression analyses to monitor the expression of C5a, C5aR, IL‐6, and TLR‐4 at the protein and mRNA levels. LPS stimulation for 48 h and treatment with W54011 for 48 h effectively inhibited inflammation and did not affect C5a expression. This study provides a basis for follow‐up studies of W54011 in dental pulp cells.

目的 应用三维有限元法探讨髓腔深度对E-Max铸瓷高嵌体修复下颌第一磨牙近中邻面(mesial-occlusal,MO)缺损的应力大小及分布情况.方法 选取1名牙周健康、牙列完整、咬合正常、左侧下颌第一磨牙形态标准的哈尔滨医科大学附属第二医院口腔颌面外科的志愿者作为建模对象,采用锥形束CT(conical beam CT,CBCT)扫描数据结合逆向工程软件建立下颌第一磨牙根管治疗后MO缺损的有限元模型,选择2种不同面覆盖方式的高嵌体,A组为全覆盖式的高嵌体,B组为保留远中边缘嵴式的高嵌体.根据髓腔制备深度分为2 mm、3 mm、4 mm、5 mm组,各组分别施加280 N的垂直载荷和105 N的舌向45度载荷,观察修复体、黏结剂层及剩余牙体组织的应力大小和分布情况.结果 A组中各结构的应力值随着髓腔深度的增加而增大,髓腔深度为2 mm时,各结构的应力值相对最小;B组在髓腔深度为5 mm时,垂直加载下,各结构应力值最小,斜向加载下,黏结剂和牙本质的应力值最大;B组与A组的应力大小相似,但斜向加载时,除牙釉质外各结构应力值均变小.结论 增加髓腔深度,全覆盖式高嵌体易脱落和折裂,而保留边缘嵴式高嵌体本身能得到有效保护,值得临床推广.

Background During the process of deep decay, when decay approaches the pulp, an immune response is triggered inside the pulp, which activates the complement cascade. The effect of complement component 5a (C5a) on the differentiation of dental pulp mesenchymal stem cells (DPSCs) is related to dentin reparation. The aim of the present study was to stimulate DPSCs with different concentrations of C5a and evaluate the differentiation of odontoblasts using dentin sialoprotein (DSP). Methods DPSCs were divided into the following six groups: (i) Control; (ii) DPSCs treated with 50 ng/ml C5a; (iii) DPSCs treated with 100 ng/ml C5a; (iv) DPSCs treated with 200 ng/ml C5a; (v) DPSCs treated with 300 ng/ml C5a; and (vi) DPSCs treated with 400 ng/ml C5a. Flow cytometry and multilineage differentiation potential were used to identify DPSCs. Mineralization induction, Real-time PCR and Western blot were conducted to evaluate the differentiation of odontoblast in the 6 groups. Result DPSCs can express mesenchymal stem cell markers, including CD105, CD90, CD73 and, a less common marker, mesenchymal stromal cell antigen-1. In addition, DPSCs can differentiate into adipocytes, neurocytes, chondrocytes and odontoblasts. All six groups formed mineralized nodules after 28 days of culture. Reverse transcription-quantitative PCR and western blotting indicated that the high concentration C5a groups expressed higher DSP levels and promoted DPSC differentiation, whereas the low concentration C5a groups displayed an inhibitory effect. Conclusion In this study, the increasing concentration of C5a, which accompanies the immune process in the dental pulp, has demonstrated an enhancing effect on odontoblast differentiation at higher C5a concentrations in vitro.

背景:在牙髓-牙本质再生过程中,牙髓内其他类型的细胞如成牙本质细胞和干细胞一直广受学者关注和研究,对于成纤维细胞的研究甚少,对牙髓成纤维细胞的研究可为今后牙髓治疗提供新的细胞来源.目的:综述牙髓成纤维细胞在牙髓-牙本质再生过程中的作用.方法:应用计算机在PubMed数据库、中国知网数据库,检索2000至2020年间关于牙髓成纤维细胞和牙髓-牙本质再生的相关文献,英文检索词为"pulp、fibroblast、dentin-pulp、regeneration、complement、C5a、C3a",中文检索词为"牙髓、成纤维细胞、牙本质-牙髓、补体、C3a、C5a",最终对56篇文献进行归纳总结.结果与结论:成纤维细胞是牙髓中数量最多的一种细胞,该细胞在正常及病理状态下对牙髓免疫反应及功能起着重要的调节作用.它们可以使补体蛋白产生具有生物活性的片段,如C3a、C5a和膜攻击复合物,同时通过分泌生长因子对修复性牙本质的合成、血管再生和神经的生长分布进行精细调控,这在牙髓-牙本质再生过程中发挥着重要作用.因此,除了成牙本质细胞和牙髓干细胞外,牙髓成纤维细胞也应该被视为诱导牙本质-牙髓再生策略中的一种重要细胞.

Oral squamous cell carcinoma (OSCC), the subtype of head and neck cancers, is notorious for its high incidence and death rate. The role of long non-coding RNAs (lncRNAs) is discovered to be significant for the canceration and cancer progression. Long intergenic non-protein coding RNA 958 (LINC00958) is discovered as a carcinogene in multiple cancers, such as gastric cancer, pancreatic cancer, and glioma, but there has been no report about how LINC00958 functions in OSCC. The objective of our study is to unfold function and mechanism investigation on LINC00958 in OSCC. First, TCGA database showed the upregulation and prognostic significance of LINC00958 in head and neck squamous carcinoma. Then, we discovered in OSCC clinical samples that LINC00958 presented high expression and predicted poor prognosis. Also, LINC00958 was elevated in OSCC cells. In vitro gain- and loss-function experiments proved that LINC00958 facilitated cell growth, retarded apoptosis, accelerated migration, and epithelial-to-mesenchymal transition (EMT) in OSCC. Mechanistically, we confirmed the cytoplasmic expression of LINC00958 in OSCC cells, and revealed that LINC00958 sequestered miR-627-5p to upregulate YBX2 expression. Rescue assays indicated that LINC00958 regulated OSCC cell proliferation, motility and EMT through YBX2. Together, we showed that LINC00958 promoted OSCC progression through miR-627-5p/YBX2 axis, indicating LINC00958 as a new prognostic marker, and provided new perspectives for molecular targeted treatment for OSCC.

Different types of oral adhesives have been widely used in the field of oral prosthodontics. In clinical practice, various adhesion systems are used to repair residual roots and crowns by fiber pile adhesion. However, insufficient adhesion strength and microleakage of adhesives may lead to clinical repair failure.This is still a common problem faced by clinicians.And different bonding system will directly affect the bond strength and edge micro leakage, binder and tooth tissue, repair materials strong lasting bond is formed between the fiber bonding, repair of post-and-core crowns, critical to the success of the effective bonding can be closed the gap between the restoration with tooth, them tightly together, maximum limit reduces the tooth body and the formation of micro leakage between fillings, so as to reduce the occurrence of secondary caries.In this paper, the adhesive strength and microleakage of the adhesive were reviewed in order to provide evidence-based medical basis for clinical selection of fiber pile adhesive system.

患牙经根管治疗后,牙体结构在"质"和"量"上均发生了显著变化,抗折性能明显下降.大量研究表明,为了保证牙体的抗折性能,应避免切割更多的残余牙齿组织,采用具有髓腔固位的嵌体比利用根管固位的桩核冠更有利.尤其在根管治疗后面积缺损过大的患牙、牙冠过短或临床牙根弯曲无法行桩核冠修复的患牙中,可采用嵌体冠等形式.本文主要从嵌体材料类型、粘接和微渗漏、嵌体固位型和应力分析等方面进行综述,旨在为临床微创美学修复无髓后牙提供参考.

Clinically, deep decay can lead to inflammation in the dental pulp. Apart from the use of various materials to sooth the inflamed pulp, there is currently no adequate treatment, and the gold standard, calcium hydroxide, that is used to cover the dentin/pulp, has limited effect. Sometimes the pulp will remain infected and cause pulpitis, and ultimately, the pulp will need to be removed. The first principle of oral treatment is to protect the pulp. Therefore, it is necessary to study the immune response and regeneration of pulp cells in conditions of deep decay. Of the terminal complement system proteins, complement 5a (C5a) has the most potent effect compared to complement 3a (C3a) and complement 4a (C4a). C5a is 20- to 2,500-fold stronger than C3a and C4a. The purpose of this study was to elucidate the association between C5a, secreted by complement activation, and the duration of inflammation. Another key goal was to detect the expression of C5a and its receptor, complement 5a receptor (C5aR). To this end, the cells were divided into 4 groups as per stimulation with lipoteichoic acid (LTA) or lipopolysaccharide (LPS) as follows: i) The 1 µg/ml LTA group; ii) the 1 µg/ml LPS group; iii) the 1 µg/ml LTA and 1 µg/ml LPS group; and iv) the PBS-only group, which served as a control. There were 5 time points for all 4 groups: 1, 2, 3, 5 and 7 days. Reverse transcription-quantitative polymerase chain reaction was used to detect the gene expression levels of C5a, C5aR and interleukin (IL)-6 at different time points. Western blot analyses was carried out to detect the expression of C5aR. Transmission electron microscopy was also conducted to assess the ultrastructural features of dental pulp cells. The gene expression trends of C5a and C5aR mRNA were identical. C5a and C5aR mRNA was highly expressed on the second day of LTA or LPS stimulation. However, in the LTA and LPS co-stimulation group, C5a and C5aR mRNA were highly expressed on both the first and second day, with higher levels on the second day. IL-6 expression decreased as time progressed in the LTA only and in the LTA + LPS co-stimulation groups. However, a peak in its expression was observed on the second day in the LPS group. On the whole, this study demonstrates that a 1 µg/ml concentration of LTA and LPS stimulates human dental pulp cells to activate the expression of C5a.

嵌体作为恢复牙体缺损的间接修复方式,具有美观、耐用以及边缘密合性良好等优势,故逐渐被患者接受.随着全瓷材料工艺与技术的革新,全瓷嵌体已广泛应用于临床,并逐渐替代了金属和树脂嵌体.对全瓷嵌体的各种特性进行研究的重要性逐渐凸显,其中嵌体材料、洞型设计、黏合剂等因素对嵌体及剩余牙体组织的影响很大.随着口腔生物力学相关研究的不断深入,三维有限元分析法作为新的准确、高效的生物力学研究方法,已广泛应用于口腔医学领域.

牙源性干细胞对口腔领域的新型细胞疗法的发展具有重要意义,引起广泛的关注.在牙体组织发育或再生过程中,牙源性干细胞也非常适合研究细胞过程.牙囊中的多能未分化细胞是牙源性干细胞的一种,这些细胞已被用于分化为成牙骨质细胞和成骨细胞的细胞过程的研究,对于牙周组织具有重要意义.该文关于信号通路、转录因子、细胞外基质蛋白对牙囊细胞成骨分化的研究作一综述.

Dental pulp stem cells(DPSC)are mesenchymal stem cells derived from neural crest,which have strong self-renewal and multidifferentiation ability. They can be removed from the tooth pulp of the extracted tooth in a relatively easy way. Accumulating evi-dence suggested that dental pulp stem cell therapy has a great potential in the treatment of diseases of the central nervous system (CNS)and retinal damage. Dental pulp stem cells can produce therapeutic effects in a number of ways,especially a series of trophic factors mediated by paracrine secretion,which is seen as the dominant mechanism for its function. In this paper,the latest evidence a-bout the repair of central nervous system injury by dental pulp stem cells,including neuroprotective effects on retinal ganglion cells and the role in optic nerve regeneration and the treatment of glaucoma,is reviewed.

Objective To study the effect of tauroursodeoxycholic acid (TUDCA) on the proliferation and osteogenic differentiation in dental pulp stem cells (DPSCs). Methods Methylthiazolyltetrazolium (MTT) assay was used to evaluate the proliferation rate of DPSCs induced by different concentrations of TUDCA(0,10,100,300nM). The activity of alkaline phosphatase (ALP) after induced by different concentration of TUDCA was measured. We observed the morphologica1 changes of DPSCs by the inverted phase contrast microscope, detected the mineralize nodules stained by Alizarin red. Results The results of MTT and ALP activity measurement showed that ALP activity increased and cell proliferation rate arise with the increase of concentration of TUDCA. Alizarin red staining showed that mineralized nodules have formed after induced by TUDCA for 28 day. Conclusion TUDCA can promote the proliferation and osteogenic differentiation of DPSCs.

It has been established that dental pulp stem cells (DPSCs) serve an important role in the restoration and regeneration of dental tissues. DPSCs are present in blood vessels and also exist in the vessel microenvironment in vivo and have a close association with endothelial cells (ECs). The present study aimed to evaluate the influence of ECs and their secretory product endothelin‑1 (ET‑1) on the differentiation of DPSCs. In the present study, cells were divided into four groups: i) a DPSC‑only control group; ii) a DPSC with ET‑1 administration group; iii) a DPSC and human umbilical vein endothelial cell (HUVEC) direct co‑culture group; and iv) a DPSC and HUVEC indirect co‑culture group using a Transwell system. Reverse transcription‑quantitative polymerase chain reaction was used to detect the expression of the odontoblastic differentiation‑associated genes, including dentin sialoprotein (DSP) and dentin matrix acidic phosphoprotein 1 (DMP‑1) at days 4, 7, 14 and 21. Alizarin Red S staining, immunofluorescence and western blot analyses were also conducted to assess the differentiation of the DPSCs in each group. The highest expression levels of odontoblastic differentiation‑associated genes were observed on day 7 and in the two co‑culture groups were increased compared with the DPSC‑only and DPSC + ET‑1 culture groups at all four time points. However, expression levels in the DPSC + ET‑1 group were not downregulated as notably as in the co‑culture groups on days 14 and 21. The Transwell group exhibited the greatest ability for odontoblastic differentiation compared with the other groups according to staining with Alizarin Red S, immunofluorescence and western blot analysis results. According to the results of the present study, the culture solution with HUVECs affected the differentiation of DPSCs. In addition, ET‑1 may promote the odontoblastic differentiation of DPSCs.

许多医学领域都在研究富血小板纤维蛋白(platelet-rich fibrin,PRF)与其他生物性材料结合应用在再生治疗中的临床优势.尽管如此,PRF在组织修复和再生方面的实际临床作用还存在许多问题.该文就当前PRF在口腔领域内再生性治疗的临床潜能作一综述.

Stem cell biology has become an important field in regenerative medicine and tissue engineering therapy since the discovery of mesenchymal stem cells .Mesenchymal stem cells can be isolated from bone marrow ,fat,umbilical cord blood and tooth tissues ,among which dental stem cells are relatively easy to obtain and have high plasticity and pluri-potency,and can be differentiated into multiple cells such as brain derived neurotrophic factors ,hepatocytes,pancreatic cells,and corneal cells,for the treatment of various diseases,such as neurological diseases,cirrhosis,diabetes and cornea blindness.Further studies on dental stem cells may bring hope to the treatment of some diseases .

Human dental pulp stem cells (hDPSCs) possess self-renewal and osteogenic differentiation properties, and have been used for orofacial bone regeneration and periodontal treatment. Aspirin has been demonstrated to enhance the regeneration of bone marrow mesenchymal stem cells (MSCs); however, the impact of aspirin on the osteogenic differentiation of hDPSCs remains unknown. In the present study, hDPSCs were characterized by flow cytometry, while their clonogenic potential and multipotency were assessed using alizarin red, Oil red O and alcian blue staining. The effect of aspirin on hDPSC viability was assessed using Cell Counting Kit-8 assay. Osteogenic capacity was examined by alkaline phosphatase activity, alizarin red staining, reverse transcription-polymerase chain reaction and western blotting. Furthermore, in vivo cranial defects were established in Sprague-Dawley rats to evaluate the effect of aspirin on hDPSC-based bone regeneration. Anorganic bovine bone was used as a bone replacement material and as the carrier for hDPSCs. New bone formation was observed through radiographic and histological analysis. The study demonstrated that hDPSCs expressed MSC markers and possessed multipotency in vitro. Aspirin was non-toxic to hDPSCs at a concentration of <= 100 mu g/ml and enhanced the osteogenesis of hDPSCs in vitro. Aspirin significantly increased hDPSC-based bone formation in the rat cranial defect model at 8 or 12 weeks post-implantation (P<0.05). The data suggested that aspirin promotes the osteogenic potential of hDPSCs in vitro and in vivo. Overall, the present study indicated that aspirin improves the bone regeneration capacity of hDPSCs.

Dental pulp stem cells (DPSCs) have been proposed as a promising source of stem cells in nerve regeneration due to their close embryonic origin and ease of harvest. Resveratrol (RSV) is a natural polyphenolic and possesses many biological functions such as anti-inflammatory activity and protection against atherosclerosis and neuroprotective activities. There is increasing evidence showing that RSV plays a pivotal role in neuron protection and neuronal differentiation. In this study, we isolated DPSCs from impacted third molars and investigated whether RSV induces neuronal differentiation of DPSCs. To avoid loss of DPSCs multipotency, all the experiments were conducted on cells at early passages. RT-PCR results showed that RSV-treated DPSCs (RSV-DPSCs) significantly increased the expression of the neuroprogenitor marker Nestin. When RSV-DPSCs were differentiated with neuronal induction media (RSV-dDPSCs), they showed a cell morphology similar to neurons. The expression of neuronal-specific marker genes Nestin, Musashi, and NF-M in RSV-dDPSCs was significantly increased. Immunocytochemical staining and Western blot analysis showed that the expression of neuronal marker proteins, Nestin, and NF-M, was significantly increased in RSV-dDPSCs. Therefore, we have shown that RSV treatment, along with the use of neuronal induction media, effectively promotes neuronal cell differentiation of DPSCs.

Objects LIM mineralization protein?1 is non?secret protein in the cell, widely existing in various tissues, which not only affects the mineralization of bone matrix but also is the important regulatory factor in the process of osteoblast differentiation and bone formation. This experiment evaluates the effect of intracellular signal transduction molecules LIM mineralization protein 1on dental pulp stem cells in vitro mineralization by detecting the process of mineralization. It can provide the reference for evaluating the LMP?1 as os?teogenesis adjustment factor and affecting bone matrix mineralization research. Methods Dental pulp stem cells were cultured in vitro, purified and then subcultured. Bone marrow mesenchymal stem cells were cultured in vitro by density gradient centrifugation method. Two kinds of cells were set up respectively into the experimental group and the control group. In the experimental group, the induction fluid was added to the culture medium, but in the control group it was not added. They were detected in 3, 5, 7 and 14 days respec?tively. Fluorescence quantitative PCR method was used to detect the culture period expression of osteogenic marker genes LIM minerali?zation protein?1、BMP?2、DSPP、COL?1、RUNX?2. The results were statistically analyzed by SPSS17.0. Results The activity of alkaline phosphatase increased and the activity of alkaline phosphatase in the experimental group was significantly higher than that in the control group at 5, 3, 7 and 14 days. The formation of mineralized nodules was seen in the experimental group at 21 days. Fluorescent quanti?tative PCR was used in both the experimental and control groups, and the expression of the experimental group was significantly higher than that of the control group. Conclusion LIM mineralization protein?1 is associated with the mineralization of dental pulp stem cells in vitro and bone marrow mesenchymal stem cells. It’ s speculated that LIM mineralization protein 1 May as the detection of dental pulp stem cells and bone marrow mesenchymal stem cells is an indicator of mineralization.

Stem cells have highly self-renewal,proliferation and differentiation potential,through cell division to maintain the size of their cell mass,and can differentiate into various tissue cells at the same time,which constitute the complex body organ.However,stem cells are rare,and the content is very low,so to solve the mesenchymal stem cell rapid proliferation is a hot issue in the research of tissue engineering.The platelet rich plasma (Platelet rich,plasma,PRP) comes from the autologous blood after centrifugation,which contains a large number of platelets and a small quantity of other blood cells.Activated PRP can influence the proliferation of stem cell through the combined and synergistic effect of growth factors,such as platelet-derived growth factor,insulin-like growth factor and transforming growth factor and PRP can overcome the defects of Heterologous rejection,and has low price,high biological safety,and fewer complications.Therefore,it is important to understand the effect of PRP on the proliferation and differentiation of stem cells.