不同浓度钙离子干预人牙周膜干细胞的增殖和成骨分化

背景：钙离子作为第二信使,调控着众多的生理过程,但在牙周膜干细胞成骨分化方面的生物学效应尚不明确。目的：探讨钙离子在人牙周膜干细胞增殖和成骨分化过程中的作用。方法：使用含钙离子浓度为0(对照组),1,2,5,10,15 mmol/L的完全培养基及成骨诱导液对第3代人牙周膜干细胞进行增殖能力检测及成骨诱导。培养1,3,5,7 d使用CCK-8检测细胞增殖能力;在成骨诱导第3,7天采用Real-Time PCR和Western blot检测RUNX2、OPN、OCN的基因及蛋白表达;成骨诱导第14天时进行茜素红染色分析人牙周膜干细胞的成骨矿化情况。结果与结论：(1)CCK-8检测结果显示,2,5 mmol/L钙离子组人牙周膜干细胞在第5,7天的增殖能力优于对照组(P <0.05);(2)体外成骨诱导7 d后,5,10 mmol/L钙离子组人牙周膜干细胞中RUNX2、OPN、OCN的表达高于对照组(P <0.05);(3)体外成骨诱导14 d后,含钙离子组人牙周膜干细胞的矿化结节数量均高于对照组(P <0.05);(4)以上结果表明,在一定浓度范围内,钙离子可促进人牙周膜干...     

大麻二酚促进人牙周膜干细胞增殖和成骨分化

背景:大麻二酚是大麻中主要的非精神活性成分,具有抗炎、抗氧化等药理活性,最近研究表明,大麻二酚能有效促进前成骨细胞和间充质干细胞成骨分化,但大麻二酚对人牙周膜干细胞增殖和成骨分化的影响尚不明确.目的:探究大麻二酚对人牙周膜干细胞增殖和成骨分化的影响.方法:采用组织块法分离培养人牙周膜干细胞,通过形态学观察、流式细胞术分...     

低频脉冲电磁场在三维支架下对人脂肪源干细胞增殖和成骨分化的影响北大核心

背景:目前已有较多实验研究证实低频脉冲电磁场在二维支架下促进多种干细胞向成骨细胞分化,但鲜有研究磁场在三维支架下对干细胞的增殖及成骨分化的影响。目的:探讨在聚已内酯3D培养支架环境下,外加低频脉冲电磁场对人脂肪源干细胞增殖及成骨分化的影响。方法:将第3代人脂肪源干细胞接种于聚已内酯3D培养支架上,分2组培养,实验组给予50 Hz、1 m T的低频脉冲电磁场干预,2 h/d,连续干预14 d;对照组不给于低频脉冲电磁场干预。培养7 d后,Live/Dead染色观察细胞存活情况;培养1,3,5,7 d后,MTT法检测细胞增殖;培养7,14 d,碱性磷酸酶染色及qR T-PCR检测细胞成骨分化水平。结果与结论:(1)Live/Dead染色:人脂肪源干细胞已长入到支架材料的空隙结构内部,在材料中可保持较高的活性;(2)细胞增殖:随着培养时间的延长,两组细胞A值均逐渐升高,实验组培养1,3 d的细胞A值高于对照组(P<0.05);(3)碱性磷酸酶染色:实验组培养7,14 d的碱性磷酸酶阳性染色强于对照组;(4)qR T-PCR检测:培养7 d时,实验组碱性磷酸酶、Ⅰ型胶原基因表达量高于对照组(P<0.01);培养14 d时,实验组骨桥蛋白、Runt相关转录因子2、碱性磷酸酶及Ⅰ型胶原的表达量均明显高于对照组(P<0.05);(5)结果表明:在聚已内酯3D培养支架环境下,外加低频脉冲电磁场可促进人脂肪源干细胞的增殖及成骨分化。     

人牙周膜干细胞与牙周膜细胞生物学特性的比较

背景：牙周膜干细胞生物作用是目前牙周病治疗研究的热点,牙周膜成纤维细胞是其分化的终末功能细胞之一,也是其主要的支持细胞,两者生物学特性的差异研究鲜有报道。 目的：比较牙周膜干细胞与牙周膜细胞生物学特性的差异。 方法：用组织块法体外对牙周膜细胞以及单细胞克隆分离纯化后的人牙周膜干细胞两种细胞分别进行显微镜下形态观察,CCK8法检测并绘制2种细胞的生长曲线。流式细胞分析比较2种细胞的细胞周期以及细胞表面标记物的表达、实时PCR对2种细胞碱性磷酸酶、增殖细胞核抗原和Scleraxis基因进行检测。 结果与结论：牙周膜干细胞与牙周膜成纤维细胞外观差别明显,人牙周膜干细胞的生长曲线培养前5 d要低于牙周膜细胞,但在5 d后明显高于牙周膜细胞。人牙周膜干细胞与牙周膜细胞的细胞周期分别为41.1%和23.9%。表面标记物检测结果显示2种细胞虽有相似的表达,但在表达率差异有有显著性意义。实时荧光定量PCR结果显示,人牙周膜干细胞在碱性磷酸酶、增殖细胞核抗原以及Scleraxis基因的表达检测均高于牙周膜细胞。表明牙周膜干细胞在成骨增殖等生物学功能上比牙周膜细胞具有更强的潜能。中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程全文链接：     

灌注型生物反应器中流速对人骨髓间充质干细胞增殖及成骨分化的影响

目的探讨在灌注型生物反应器中,大段磷酸三钙(β-TCP)载体内不同流速对人骨髓间充质干细胞(humanmesenchymal stem cells,hMSCs)增殖及成骨分化的影响。方法利用灌注型生物反应器对复合了hMSCs的大段β-TCP载体分别以3、6、9 mL/min的流速培养15 d,通过葡萄糖消耗量、细胞活力(MTT比色法)以及扫描电镜(SEM)观测细胞在载体内的增殖情况;以Real-time PCR检测成骨分化相关基因的表达。结果各组葡萄糖的日消耗量随培养时间的延长而增加,初期9 mL/min组细胞增殖明显快于3、6 mL/min组(P<0.001),但是灌注培养后期6 mL/min组的细胞增殖快于9、3 mL/min组(P<0.05)。灌注培养15 d后,6 mL/min组细胞活力显著高于3、9 mL/min组(P<0.001)。SEM观察发现3组β-TCP载体内均形成复合细胞层,3 mL/min组细胞层呈疏松的簇状,6 mL/min组复合细胞层部分呈膜片状,9 mL/min组复合细胞层多数呈膜片状。在进行成骨诱导灌注培养15 d后,6、9 mL/min组碱性磷酸酶(ALP)及骨桥蛋白(OP)的表达均显著高于3 mL/min组(P<0.01);3、6 mL/min组骨钙素(OC)的表达基本相同(P>0.05),而9 mL/min组OC的表达量则显著高于另外两组(P<0.001)。结论在利用灌注型生物反应器对hMSCs进行灌注培养的早期,9 mL/min的流速最有利于hMSCs的增殖,而晚期6 mL/min的流速最有利于hMSCs的增殖。β-TCP载体内流体剪切应力(flow shear stress,FSS)随灌注流速的增加而增加,适当的FSS可促进细胞外基质的合成和分布,并增加成骨相关基因的表达。     

转化生长因子β3与骨形成蛋白2对牙髓干细胞增殖和成骨分化的影响

背景:转化生长因子β3与骨形成蛋白2对牙髓干细胞的增殖和成骨向分化能力对比相关研究甚少.目的:探讨相同质量浓度转化生长因子β3与骨形成蛋白2对牙髓干细胞成骨向分化的作用及其初步机制.方法:采用酶解组织块法从新西兰大白兔牙髓组织中分离培养牙髓干细胞;将第3代牙髓干细胞分为空白组、骨形成蛋白2(80μg/L)组和转化生长因...     

维甲酸诱导小型猪牙周膜干细胞的体外成骨

背景：近期研究发现维甲酸对胚胎干细胞及多种成体干细胞具有成骨方向诱导的作用。 目的：观察维甲酸对小型猪牙周膜干细胞体外成骨作用的影响。 方法：采用组织块法获得小型猪牙周膜细胞,有限稀释法纯化小型猪牙周膜干细胞,免疫荧光法检测STRO-1、免疫细胞化学法检测波形蛋白、角蛋白鉴定小型猪牙周膜干细胞。CCK8法测定小型猪牙周膜干细胞增殖曲线,检测小型猪PDLSC克隆形成率。使用维甲酸对第3代小型猪牙周膜干细胞进行成骨诱导,茜素红染色检测矿化结节,免疫细胞化学方法检测成骨相关基因骨桥蛋白、骨钙素、Ⅰ型胶原、Ⅲ型胶原的表达。 结果与结论：有限稀释法分离纯化小型猪牙周膜干细胞STRO-1,波形蛋白阳性表达,角蛋白阴性表达,克隆形成率为2.8%,维甲酸诱导14 d后,碱性磷酸酶染色阳性,诱导21 d后茜素红染色阳性,免疫细胞化学法检测骨桥蛋白、骨钙素、Ⅰ型胶原阳性表达,Ⅲ型胶原阴性表达。结果表明小型猪牙周膜干细胞能够被维甲酸诱导向成骨样细胞分化。     

P2X7受体对牙周膜干细胞成骨分化的作用

背景：研究影响牙周膜干细胞成骨分化的相关离子通道。 目的：初步观察P2X7受体在人牙周膜干细胞成骨分化中的作用。 方法：分离培养人牙周膜干细胞,分为4组,分别添加成骨诱导液和100 nmol/L三磷酸腺苷、成骨诱导液、100 nmol/L三磷酸腺苷及普通培养基培养,于成骨诱导7,14 d,利用茜素红染色检测成骨效果,qRT-PCR,Western blot检测OCN、Runx2以及P2X7受体的表达。 结果与结论：茜素红染色显示成骨诱导液+三磷酸腺苷组牙周膜干细胞的成骨效果在7 d时显著好于成骨诱导液组,但在14 d时成骨诱导液组成骨效果显著好于成骨诱导液+三磷酸腺苷组(P < 0.05);在成骨诱导7 d时,qRT-PCR结果显示成骨诱导液+三磷酸腺苷组Runx2,OCN mRNA的表达达到峰值,而14 d时明显降低(P < 0.05)。成骨诱导7 d时,成骨诱导液+三磷酸腺苷组P2X7受体mRNA的表达量明显高于三磷酸腺苷组(P < 0.05),成骨诱导液组和对照组未见P2X7受体mRNA的表达,成骨诱导14 d时,成骨诱导液+三磷酸腺苷组表达量下降,明显低于三磷酸腺苷组,差异有显著性意义(P < 0.05)。Western blot结果显示,成骨诱导7 d时,成骨诱导液+三磷酸腺苷组P2X7受体的表达达到峰值,高于三磷酸腺苷组,而成骨诱导液组表达量低,对照组几乎没有表达;成骨诱导14 d时,成骨诱导液+三磷酸腺苷组P2X7受体的表达比7 d时明显下降,而三磷酸腺苷组P2X7受体的表达比7 d时有所增加。结果表明外源性三磷酸腺苷可在牙周膜干细胞成骨诱导前7 d明显提高成骨效果,但在7 d后,抑制成骨诱导效果,三磷酸腺苷可以激活牙周膜干细胞P2X7受体表达,且P2X7受体的表达与牙周膜干细胞的成骨效果正相关。中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程全文链接：     

牙周膜干细胞成骨分化中细胞因子的作用

背景：牙周膜干细胞是牙周组织中的成体干细胞,具有高度增殖、自我更新能力和多分化潜能。促进牙周膜干细胞向成骨细胞分化有助于牙周疾病的治疗。 目的：观察胰岛素样生长因子1和成纤维细胞生长因子2对牙周膜干细胞向成骨细胞分化的影响。 方法：采用胶原酶消化人牙周膜组织,获得牙周膜干细胞,经体外鉴定、扩增后,通过倒置显微镜、苏木精-伊红染色、流式细胞仪对牙周膜干细胞进行生物学检测。分别在成骨细胞诱导培养液中加入成骨诱导液(对照组)及胰岛素样生长因子1和成纤维细胞生长因子2持续诱导7,14 d后,进行碱性磷酸酶染色、碱性磷酸酶活性检测,以及茜素红染色,并用实时定量PCR法检测向成骨细胞分化的标志性基因的表达情况。 结果与结论：胰岛素样生长因子1刺激组的碱性磷酸酶活性以及钙化结节明显高于对照组,Runx2、Alp、col-1的mRNA呈高表达;成纤维细胞生长因子2刺激组的碱性磷酸酶活性以及钙化结节也高于对照组,Runx2、Alp、col-1的mRNA表达量也高于对照组。提示胰岛素样生长因子1和成纤维细胞生长因子2在不同程度上促进体外培养的牙周膜干细胞向成骨细胞方向分化。     

牙周膜干细胞诱导骨髓间充质干细胞的牙向分化

目的 探讨牙周膜干细胞(PDLSC)诱导骨髓间充质干细胞(BMMSC)牙向分化的机制,为联合应用PDLSC和BMMSC再生牙周复合体提供实验依据.方法应用Transwell(R)小室法间接联合培养小型猪PDLSC和BMMSC,根据二者混合比例随机分为3组.A组:P∶M=10∶1共培养组;B组:P∶M=1∶1共培养;C组:P:M=1:10共培养,单独PDLSC和BMMSC培养组分别为阳性和阴性对照组.培养14 d,应用免疫荧光染色和实时定量PCR(qRT-PCR)分别检测scleraxis、osteocalcin(OCN)、osterix(OSX)、细胞外基质磷酸糖蛋白(MEPE)蛋白和mRNA表达情况,以判定PDLSC诱导BMMSC成牙的最佳配比比例.结果 免疫荧光染色和qRT-PCR结果均显示scleraxis、OCN和OSX相对mRNA表达水平在A、B、C组间没有统计学差异(P>0.05),但相对MEPE mRNA表达水平在A组却明显高于B组和C组(P＜0.01).结论 联合培养可促进BMMSC获得不同程度的PDLSC特性,且少量的PDLSC同样可以促进BMMSC获得牙源性干细胞特性.     

Influence of different intensities of vibration on proliferation and differentiation of human periodontal ligament stem cells

Introduction

To understand the effects of low-magnitude, high-frequency (LMHF) mechanical vibration at different intensities on human periodontal ligament stem cell (hPDLSC) proliferation and osteogenic differentiation.

Material and methods

The effect of vibration on hPDLSC proliferation, osteogenic differentiation, tenogenic differentiation and cytoskeleton was assessed at the cellular, genetic and protein level.

Results

The PDLSC proliferation was decreased after different magnitudes of mechanical vibration; however, there were no obvious senescent cells in the experimental and the static control group. Expression of osteogenesis markers was increased. The expression of alkaline phosphatase (ALP) and osteocalcin (OCN) mRNA was up-regulated at 0.1 g, 0.3 g, 0.6 g and 0.9 g magnitude, with the peak at 0.3 g. The type I collagen (Col-I) level was increased after vibration exposure at 0.1 g, 0.3 g, and 0.6 g, peaking at 0.3 g. The expression levels of both mRNA and protein of Runx2 and osterix (OSX) significantly increased at a magnitude of 0.1 g to 0.9 g, reached a peak at 0.3 g and then decreased slowly. The scleraxis, tenogenic markers, and mRNA expression decreased at 0.05 g, 0.1 g, and 0.3 g, and significantly increased at 0.6 g and 0.9 g. Compared with the static group, the F-actin stress fibers of hPDLSCs became thicker and clearer following vibration.

Conclusions

The LMHF mechanical vibration promotes PDLSC osteogenic differentiation and implies the existence of a magnitude-dependent effect of vibration on determining PDLSC commitment to the osteoblast lineage. Changes in the cytoskeleton of hPDLSCs after vibration may be one of the mechanisms of the biological effects.     

Periostin promotes migration,proliferation, and differentiation of human periodontal ligament mesenchymal stem cells

Overview: Periostin (POSTN) is critical to bone and dental tissue morphogenesis, postnatal development, and maintenance; however, its roles in tissue repair and regeneration mediated by human periodontal ligament mesenchymal stem cells (PDLSCs) remain unclear. The present study was designed to evaluate the effects of POSTN on hPDLSCs in vitro. Materials and Methods: hPDLSCs were isolated and characterized by their expression of the cell surface markers CD44, CD90, CD105, CD34, and CD45. Next, 100 ng/mL recombinant human POSTN protein (rhPOSTN) was used to stimulate the hPDLSCs. Lentiviral POSTN shRNA was used to knockdown POSTN. The cell counting kit-8 (CCK8) and scratch assay were used to analyze cell proliferation and migration, respectively. Osteogenic differentiation was investigated using an alkaline phosphatase (ALP) activity assay, alizarin staining, and quantitative calcium analysis and related genes/protein expression assays. Results: Isolated hPDLSCs were positive for CD44, CD90, and CD105 and negative for CD34 and CD45. In addition, 100 ng/mL rhPOSTN significantly accelerated scratch closure, and POSTN-knockdown cells presented slower closure at 24 h and 48 h. Furthermore, the integrin inhibitor Cilengitide depressed the scratch closure that was enhanced by POSTN at 24 h. The CCK8 assay showed that 100 ng/mL rhPOSTN promoted hPDLSC proliferation. Moreover, 100 ng/mL rhPOSTN increased the expression of RUNX2, OSX, OPN, OCN, and VEGF and enhanced ALP activity and mineralization. POSTN silencing decreased the expression of RUNX2, OSX, OPN, OCN, and VEGF and inhibited ALP activity and mineralization. Conclusions: POSTN accelerated the migration, proliferation, and osteogenic differentiation of hPDLSCs.     

地塞米松诱导牙周膜干细胞的定向成骨分化及凋亡

背景：牙周膜干细胞是一类起源于牙组织的成体干细胞,具有良好的成骨分化能力,有望在骨组织工程中得到应用。 目的：观察成骨诱导液对牙周膜干细胞成骨分化能力及细胞早期凋亡的影响。 方法：从原代牙周膜组织中分离得到牙周膜干细胞,以1×10⁴/cm²浓度铺板后开始诱导。利用1,10,100 nmol/L地塞米松、β-磷酸甘油钠、维生素C为成骨诱导剂,以碱性磷酸酶活性检测、茜素红矿化结节染色、荧光定量PCR等方法对细胞成骨情况进行鉴定,采用AnnexinV/PI双染法检测细胞凋亡情况。 结果与结论：地塞米松可有效诱导牙周膜干细胞成骨分化,可显著提高碱性磷酸酶活性,促进茜素红矿化结节形成,提高成骨相关基因骨粘连蛋白及Ⅰ型胶原表达。根据碱性磷酸酶活性和矿化结节实验结果,地塞米松的成骨诱导具有浓度梯度效应,其中100 nmol/L 地塞米松具有最佳成骨诱导能力。细胞凋亡结果提示,地塞米松诱导的成骨分化具有一定促凋亡作用,可诱导牙周膜细胞的早期凋亡。     

Cyclic tension promotes osteogenic differentiation in human periodontal ligament stem cells

Orthodontic forces result in alveolar bone resorption and formation predominantly on the pressure and tension sides of the tooth roots, respectively. Human periodontal ligament stem cells (PDLSCs) have demonstrated the capacity to differentiate into osteoblasts, and they play important roles in maintaining homeostasis and regenerating periodontal tissues. However, little is known about how PDLSCs contribute to osteoblastogenesis during orthodontic tooth movement on the tension side. In this study, we applied a 12% cyclic tension force to PDLSCs cultured in osteoinductive medium. The osteogenic markers Runx2, ALP, and OCN were detected at the mRNA and protein levels at different time points using real-time PCR and western blot analyses. We discovered that the mRNA and protein levels of Runx2, ALP and OCN were significantly up-regulated after 6, 12 and 24 hours of mechanical loading on PDLSCs compared to levels in unstimulated PDLSCs (P < 0.05). This study demonstrates, for the first time, the effects of mechanical tensile strain on the osteogenic differentiation of PDLSCs, as examined with a Flexcell FX-4000T Tension Plus System. Our findings suggested that cyclic tension could promote the osteogenic differentiation of PDLSCs. Furthermore, the effects of orthodontic force on alveolar bone remodeling might be achieved by PDLSCs.     

Up-regulated osteogenic transcription factors during early response of human periodontal ligament stem cells to cyclic tensile strain

Introduction

As one group of periodontal ligament (PDL) cells, human periodontal ligament stem cells (hPDLSCs) have been isolated and identified as mesenchymal adult stem cells (MSCs) since 2004. It has been well accepted that PDL sensitively mediates the transmission of stress stimuli to the alveolar bone for periodontal tissue remolding. Besides, the direction of MSCs differentiation has been verified regulated by mechanical signals. Therefore, we hypothesized that tensile strain might act on hPDLSCs differentiation, and the early response to mechanical stress should be investigated.

Material and methods

The hPDLSCs were cultured in vitro and isolated via a magnetic activated CD146 cell sorting system. After investigation of surface markers and other experiments for identification, hPDLSCs were subjected to cyclic tensile strain at 3,000 µstrain for 3 h, 6 h, 12 h, and 24 h, without addition of osteogenic supplements. In the control groups, the cells were cultured in similar conditions without mechanical stimulation. Then osteogenic related genes and proteins were analyzed by RT-PCR and western blot.

Results

Cyclic tensile strain at 3,000 µstrain of 6 h, 12 h, and 24 h durations significantly increased mRNA and protein expressions of Satb2, Runx2, and Osx, which were not affected in unloaded hPDLSCs.

Conclusions

We indicate that hPDLSCs might be sensitive to cyclic tensile strain. The significant increase of Runx2, Osx and Satb2 expressions may suggest an early response toward osteogenic orientation of hPDLSCs.     

Nitric oxide production during the osteogenic differentiation of human periodontal ligament mesenchymal stem cells

The critical tissues that require regeneration in the periodontium are of mesenchymal origin; therefore, the ability to identify, characterize and manipulate mesenchymal stem cells within the periodontium is of considerable clinical significance. In particular, recent findings suggest that periodontal ligament cells may possess many osteoblast-like properties. In the present study, periodontal ligament mesenchymal stem cells obtained from healthy volunteers were maintained in culture until confluence and then induced to osteogenic differentiation. Intracellular calcium ([Ca²⁺]_i) concentration and nitric oxide, important signalling molecules in the bone, were measured along with cell differentiation. Alkaline phosphatase activity was assayed and bone nodule-like structures were evaluated by means of morphological and histochemical analysis. Our results showed that the periodontal ligament mesenchymal stem cells underwent an in vitro osteogenic differentiation, resulting in the appearance of active osteoblast-like cells together with the formation of calcified deposits. Differentiating cells were also characterized by an increase of [Ca²⁺]_i and nitric oxide production. In conclusion, our data show a link between nitric oxide and the osteogenic differentiation of human periodontal ligament mesenchymal stem cells, thus suggesting that local reimplantation of expanded cells in conjugation with a nitric oxide donor could represent a promising method for treatment of periodontal defects.     

人牙周膜干细胞来源外泌体干预成骨细胞MC3T3-E1的增殖和分化

背景:人牙周膜干细胞具有较强的成骨分化能力,人牙周膜干细胞来源外泌体作为牙周膜干细胞分泌的主要成分,对成骨细胞MC3T3-E1增殖和成骨分化的影响尚不明确。目的:探讨人牙周膜干细胞来源外泌体对MC3T3-E1细胞增殖和分化的影响。方法:采用酶消化法分离及培养人牙周膜干细胞,超速离心法提取人牙周膜干细胞来源外泌体,通过透射电镜、粒径分析及Western blot方法对人牙周膜干细胞来源外泌体进行鉴定;CCK8法检测不同质量浓度人牙周膜干细胞来源外泌体对MC3T3-E1细胞增殖的影响,茜素红染色观察100 mg/L人牙周膜干细胞来源外泌体对MC3T3-E1细胞成骨矿化的影响,Western blot检测100 mg/L人牙周膜干细胞来源外泌体干预前后MC3T3-E1细胞内MEK和ERK的磷酸化水平。结果与结论:①透射电镜观察可见外泌体为脂质双分子层形成的囊泡结构,粒径检测显示外泌体直径分布在50-120 nm,集中在79.86 nm,Western blot检测结果显示提取的外泌体中含有CD81,CD63,TSG101的表达;②与对照组相比,人牙周膜干细胞来源外泌体对MC3T3-E1细胞的增殖具有促进作用,且作用呈剂量依赖性;③与对照组相比,人牙周膜干细胞来源外泌体组MC3T3-E1细胞能够形成更多的钙结节;与对照组相比,人牙周膜干细胞来源外泌体组MC3T3-E1细胞内p-MEK及p-ERK蛋白表达量升高;④结果表明,人牙周膜干细胞来源外泌体可以显著促进MC3T3-E1增殖和成骨分化,推测可能与其激活MEK/ERK信号通路有关。