牙髓干细胞来源的凋亡囊泡对骨髓间充质干细胞增殖和成骨分化能力的影响

王鑫源; 姚文德; 李彦; 习佳飞; 岳文; 周军年; 贺慧霞

doi:10.12435/j.issn.2095-5227.2024.072

牙髓干细胞来源的凋亡囊泡对骨髓间充质干细胞增殖和成骨分化能力的影响

Effect of apoptotic vesicles derived from dental pulp stem cells on proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells

摘要

摘要:
背景间充质干细胞来源的凋亡囊泡具有促进骨组织再生的作用，目前关于人牙髓干细胞凋亡囊泡(apoptotic vesicles derived from human dental pulp stem cells，hDPSC-apovs)用于骨组织再生的研究尚未见报道。
目的探讨hDPSC-apovs对小鼠骨髓间充质干细胞(mouse bone marrow mesenchymal stem cells，mBMSC)增殖和成骨分化能力的影响。
方法原代培养、分离和鉴定hDPSC，采用星孢素(staurosporine，STS)诱导细胞凋亡，高速离心法分离hDPSC-apovs并鉴定。将mBMSC与不同浓度的hDPSC-apovs(0 μg/mL、0.2 μg/mL、1μg/mL 、5 μg/mL、25 μg/mL)进行共培养，CCK-8法检测不同浓度hDPSC-apovs对mBMSC增殖能力的影响。mBMSC与不同浓度的hDPSC-apovs在成骨诱导培养基中共培养 7 d后，茜素红染色分析钙结节形成，qPCR检测各组mBMSC的成骨分化相关基因Alp、Runx2、Spp1、Bglap的表达。
结果与不含hDPSC-apovs的对照组相比，0.2 μg/mL、l μg/mL浓度hDPSC-apovs均能够促进mBMSC的增殖(P ＜0.05)，而5 μg/mL、25 μg/mL浓度hDPSC-apovs则抑制mBMSC的增殖(P ＜0.05)。各浓度hDPSC-apovs处理组的钙结节形成量均高于对照组(P ＜0.05)；qPCR结果显示，5 μg/mL、25 μg/mL组Alp、Runx2、Spp1、Bglap表达均上调(P ＜0.05)，1 μg/mL组只有Alp和Runx2表达上调，0.2 μg/mL组仅有Alp表达水平上调(P ＜0.05)。
结论在本研究浓度范围内，低浓度的hDPSC-apovs对mBMSC增殖具有促进作用，高浓度则表现为抑制作用；hDPSC-apovs对于mBMSC的成骨分化有一定促进作用，且不同浓度hDPSC-apovs对成骨相关基因表达的影响不同。

Abstract:
Background Apoptotic vesicles (apovs) derived from mesenchymal stem cells play a role in promoting bone tissue regeneration, while studies have not yet been reported on the application of apoptotic vesicles derived from human dental pulp stem cells (hDPSC-apovs) in bone tissue regeneration.
Objective To explore the effects of hDPSC-apovs on the proliferation and osteogenic differentiation of mouse bone marrow mesenchymal stem cells (mBMSC).
Methods Primary hDPSC was isolated and identified. The hDPSC was induced to apoptosis by staurosporine, then hDPSC-apovs were isolated by high-speed centrifugation. mBMSC were co-cultured with different concentrations of hDPSC-apovs (0, 0.2, 1, 5, and 25 μg/mL), the effect of hDPSC-apovs on mBMSC proliferation was analyzed by CCK-8 assay. Besides, mBMSC were cultured in osteogenic medium containing different concentrations of hDPSC-apovs for 7 d, then the calcium nodule forming ability was analyzed by alizarin red staining and the mRNA expression levels of Alp, Runx2, Spp1, and Bglap were analyzed by qPCR.
Results Compared with the control group, 0.2 and l μg/mL hDPSC-apovs promoted the proliferation of mBMSC (P < 0.05), while 5 and 25 μg/mL hDPSC-apovs inhibited the proliferation of mBMSC (P < 0.05). The calcium nodule forming ability of mBMSC in the groups in which mBMSC was treated by different concentrations of hDPSC-apovs was significantly higher than that in the control group (P < 0.05). Compared with the control group, the expression levels of Alp, Runx2, Spp1, and Bglap were up-regulated in the 5 and 25 μg/mL hDPSC-apovs groups, Alp and Runx2 were significantly up-regulated in the 1 μg/mLhDPSC-apovs group, but only Alp was upregulated in the 0.2 μg/mL hDPSC-apovs group (P < 0.05).
Conclusion Within the concentration range of this study, hDPSC-apovs may promote the proliferation of mBMSC at low concentrations and have an inhibitory effect at high concentrations. hDPSC-apovs can promote the osteogenic differentiation of mBMSC, and the effects on the expression of osteogenesis-related genes may be concentration-dependent.

HTML全文

参考文献(21)

施引文献

资源附件(0)