纳秒脉冲电场刺激促进人真皮成纤维细胞成骨的实验研究  

Promoting Osteogenesis of Human Dermal Fibroblasts Stimulated by Nanosecond Pulsed Electric Field:An Experimental Study

作　　者：来经天 王泽伟 董延钊 姚成俊 周海英 Ahmad Alhaskawi 李鹏飞[2] 涂田 陈永刚 Sahar Ahmed Abdalbary 卢荟[1] LAI Jingtian;WANG Zewei;DONG Yanzhao;YAO Chengjun;ZHOU Haiying;Ahmad Alhaskawi;LI Pengfei;TU Tian;CHEN Yonggang;Sahar Ahmed Abdalbary;LU Hui(Division of Orthopedics,The First Affiliated Hospital,Zhejiang University School of Medicine,Hangzhou Zhejiang 310003,China;Division of Plastic and Aesthetic,The First Affiliated Hospital,Zhejiang University School of Medicine,Hangzhou Zhejiang 310003,China;Zhejiang Provincial Key Laboratory of Pulsed Power Translational Medicine,Hangzhou Zhejiang 311100,China;Department of Orthopaedic Physical Therapy,Faculty of Physical Therapy,Nahda University in Beni Suef,Beni Suef 2711860,Egypt)

机构地区：[1]浙江大学医学院附属第一医院骨科,浙江杭州310003 [2]浙江大学医学院附属第一医院整形美容科,浙江杭州310003 [3]浙江省脉冲电场技术医学转化重点实验室,浙江杭州311100 [4]埃及贝尼苏韦夫纳赫达大学物理治疗学院骨科物理治疗系,埃及贝尼苏韦夫2711860

出　　处：《中国医疗设备》2023年第5期20-24,43,共6页China Medical Devices

基　　金：国家自然科学基金(82027803,82070516,82102183)。

摘　　要：目的 探究超短纳秒脉冲电场(Nanosecond Pulsed Electric Field,nsPEF)是否能够促进成纤维细胞向成骨细胞的表型转化。方法 用0、5 kV/cm的场强刺激成纤维细胞,观察其在普通培养基和三向分化诱导培养基中生长情况,蛋白印迹测定细胞内调控生长及分化Hippo通路下游信号分子水平,并通过体内注射细胞-凝胶复合体进行裸鼠体内成骨实验。结果 5kV/cm nsPEF促进了成纤维细胞的增殖,并促进了成骨、成软骨分化,但对成脂分化影响不显著。低能量脉冲电场刺激抑制了Hippo通路下游各信号分子磷酸化水平,促进了裸鼠体内成骨,灰度分析显示,YAP1及LATS1的磷酸化蛋白与非磷酸化蛋白比值在5 kV/cm的脉冲电场刺激后明显降低,提示HIPPO信号通路受到抑制(P<0.05)。结论 低强度nsPEF刺激有效促进了成纤维细胞的增殖和表型转化,并在裸鼠模型上证实了可行性和有效性。Objective To explore whether nanosecond pulsed electric field can promote the phenotypic transformation of fibroblasts to osteoblasts.Methods Fibroblasts were stimulated with the field strength of 0 and 5 kV/cm.The growth was observed in normal medium and three-way differentiation induction medium.The levels of downstream molecules of Hippo pathway in regulating cell growth were measured by Western blotting.Three-dimensional osteogenesis in nude mice was carried out by injecting cell-gel complex in vivo.Low energy pulsed electric field stimulation inhibited the phosphorylation levels of signal molecules downstream of Hippo pathway and promoted bone formation in nude mice.Results The 5 kV/cm of nanosecond pulsed electric field promoted fibroblast proliferation,osteogenic and chondrogenic differentiation,but had no significant effect on lipogenic differentiation.Low intensity pulsed electric field stimulation inhibited the phosphorylation levels of signal molecules downstream of Hippo pathway and promoted osteogenesis in nude mice.Gray analysis showed that the ratio of phosphorylated and non-phosphorylated proteins of YAP1 and LATS1 significantly decreased after 5 kV/cm pulsed electric field stimulation,suggesting that HIPPO signaling pathway was inhibited(P<0.05).Conclusion Low intensity nanosecond pulsed electric field stimulation can effectively promote the proliferation and phenotypic transformation of fibroblasts,which is proved to be feasible and effective in nude mouse model.

关 键 词：成骨细胞 纳秒脉冲电场 人真皮成纤维细胞 成骨试验 

分 类 号：R197.39[医药卫生—卫生事业管理]