摘要
目的:基于干细胞的牙髓再生组织工程技术已被视为治疗牙髓疾病的新技术,而干细胞的低存活率无疑会降低牙髓再生治疗的成功率。源自牙髓干细胞(dental pulp stem cells, DPSCs)的条件培养基(conditioned medium, CM)富含对组织再生至关重要的多种活性因子,从而被视为强有力的再生治疗工具。本研究旨在探索DPSCs-CM在牙髓再生中的作用及相关机制。方法:体外细胞实验中,以1,3和5天的DPSCs-CM刺激DPSCs,使用CCK-8(cell counting kit-8)检测其对DPSCs增殖的影响,通过碱性磷酸酶染色及茜素红染色观察碱性磷酸酶含量及矿化沉积水平,使用定量逆转录聚合酶链反应(quantitative real-time polymerase chain reaction, qRT-PCR)检测细胞成骨标志物基因表达水平,使用体外血管形成实验观察血管化形成水平。在裸鼠模型中,将人牙根片段植入裸鼠皮下,通过组织学评估观察牙髓再生情况。同时,通过分析DPSCs-CM的蛋白质谱深入探究其在牙髓再生过程中对细胞分化的蛋白调控机制。结果:体外实验中,DPSCs-CM不仅促进DPSCs的增殖,同时还促进了其牙源性分化性能;体内半原位牙髓再生动物模型中,相比对照组,经过4周诱导的DPSCs+5 d CM组产生了更为成熟的牙髓样结构;蛋白质谱分析显示,DPSCs-CM含有更多的牙源性和血管生成因子。结论:DPSCs-CM积极促进DPSCs增殖、成牙本质向分化及血管形成,这些发现表明DPSCs-CM在牙髓组织再生中具有潜在应用价值。
Objective:To explore the role of dental pulp stem cells conditioned medium(DPSCs-CM)in dental pulp regeneration and related mechanisms.Methods:DPSCs were stimulated with DPSCs-CM for 1,3,and 5 days,and its effect on DPSCs proliferation was detected using cell counting kit-8(CCK-8).Alkaline phosphatase(ALP)content and mineralization deposition level were observed by ALP staining and alizarin red staining.The expression level of cellular osteogenic marker genes was detected using qRT-PCR.The formation of vascularization was observed using the in vitro angiogenic assay.In the nude mouse model,human tooth root fragments were implanted subcutaneously in nude mice,and pulp regeneration was observed by histologic evaluation.Meanwhile,the protein regulation mechanism of DPSCs-CM on cell differentiation during dental pulp regeneration was deeply explored by analyzing its protein profile.Results:DPSCs-CM not only promoted the proliferation of DPSCs,but also their odontogenic differentiation properties.Compared with the control group,the DPSCs+5 d CM group induced after 4 weeks produced a more mature pulp-like structure.DPSCs-CM contained more odontogenic and angiogenic factors.Conclusion:DPSCs-CM positively promoted DPSCs proliferation,dentinogenic differentiation,and angiogenesis.These findings suggest a potential benefit of DPSCs-CM in the regeneration of dental pulp tissue.
作者
高静
王雨涵
袁莎莎
陈瑶
李璐
申静
GAO Jing;WANG Yuhan;YUAN Shasha;CHEN Yao;LI Lu;SHEN Jing(Department of International VIP Dental Clinic,Tianjin Stomatological Hospital,School of Medicine,Nankai University,Tianjin 300041,China;Tianjin Key Laboratory of Oral and Maxillofacial Function Reconstruction,Tianjin 300041,China;Department of Operative Dentistry and Endodontics,Tianjin Stomatological Hospital,School of Medicine,Nankai University,Tianjin 300041,China)
出处
《口腔医学研究》
北大核心
2025年第8期664-671,共8页
Journal of Oral Science Research
基金
天津市自然科学基金重点项目(编号:24JCZDJC00880)
天津市卫生健康委员会科技项目(编号:TJWJ2021MS039)。
关键词
条件培养基
牙髓干细胞
牙源性分化
牙髓再生
conditioned medium
dental pulp stem cells
odontogenic differentiation
pulp regeneration
作者简介
高静(1989-),女,河北人,硕士,主治医师,研究方向:牙髓病根尖周病的基础和临床研究;通信作者:申静,E-mail:shenjing611@163.com。
