静压力作用下三维培养人牙周膜干细胞促进RAW264.7细胞的破骨向分化  

Three-dimensional cultured periodontal ligament stem cells promote osteoclastic differentiation of RAW264.7 under static compression

作　　者：韩行 李雯雯 马文盛[1] Han Xing;Li Wenwen;Ma Wensheng(Stomatological Hospital of Hebei Medical University,Department of Orthodontics of Stomatological Hospital of Hebei Medical University,Key Laboratory of Stomatology in Hebei Province,Clinical Medical Research Center of Stomatology in Hebei Province,Shijiazhuang 050000,Hebei Province,China)

机构地区：[1]河北医科大学口腔医院,河北医科大学口腔医院口腔正畸科,河北省口腔医学重点实验室,河北省口腔疾病临床医学研究中心,河北省石家庄市050000

出　　处：《中国组织工程研究》2023年第24期3810-3817,共8页Chinese Journal of Tissue Engineering Research

基　　金：河北省教育厅科学研究重点项目(ZD2017054),项目负责人:马文盛;河北省医学适用技术跟踪项目(GZ2021038),项目负责人:马文盛。

摘　　要：背景:牙周膜干细胞作为种子细胞,其成骨潜能成为研究热点,而牙周膜干细胞对破骨细胞活动的调控研究较少,有待进一步探索。目的:研究持续静压力作用下三维培养人牙周膜干细胞对RAW264.7细胞破骨向分化的影响。方法:取第3,4代人牙周膜干细胞进行三维培养,随机分为13组,采用FLEXCELL5000C加压仪器,分别施加5 kPa(2,6,12 h),25 kPa(2,6,12 h),45 kPa(2,6,12 h),65 kPa(2,6,12 h)的持续静压力,第13组不加压。将细胞上清作为条件培养基按照一定比例作用于RAW264.7细胞,5 d后抗酒石酸酸性磷酸酶染色及qPCR检测破骨相关基因抗酒石酸酸性磷酸酶、组织蛋白酶K、基质金属蛋白酶9的表达。结果与结论:(1)三维培养下加压实验组与不加压对照组支架形态稳定,细胞状态良好;(2)压力加载组人牙周膜干细胞条件培养基促进RAW264.7细胞的破骨分化;(3)在一定范围内,促破骨作用与力值加载条件呈正相关,45 kPa,12h组促RAW264.7细胞破骨分化作用最强;65 kPa,12 h组的促破骨能力减弱;(4)结果表明,静压力作用下三维培养的牙周膜干细胞通过旁分泌途径促进RAW264.7的破骨分化,力值加载条件与促破骨作用具有相关性。BACKGROUND:The osteogenic potential of periodontal ligament stem cells(as seed cells)has become a research hotspot.The regulation of periodontal ligament stem cells on osteoclast activity is less studied and deserves further exploration.OBJECTIVE:To investigate the effect of three-dimensional cultured periodontal ligament stem cells under continuous static compression on the osteoclastic differentiation of RAW264.7 cells.METHODS:Periodontal ligament stem cells at passages 3 and 4 were taken for three-dimensional culture and randomly divided into 13 groups.Continuous static compression of 5 kPa(2,6,and 12 hours),25 kPa(2,6,and 12 hours),45 kPa(2,6,and 12 hours),and 65 kPa(2,6,and 12 hours)was applied by FLEXCELL 5000C,and group 13 with no compression.The cell supernatant was used as a conditioned medium to act on RAW264.7 cells.The expression of osteoclast differentiation genes tartrate resistant acid phosphatase,cathepsin K,and matrix metalloproteinase 9 was detected by qPCR and tartrate resistant acid phosphatase staining after 5 days.RESULTS AND CONCLUSION:(1)The scaffold morphology was stable and the cell status was good in the compression experimental group and the noncompression control group under three-dimensional culture.(2)The conditioned medium of periodontal ligament stem cells in the compression loading group promoted the osteoclastic differentiation of RAW264.7 cells.(3)Within a certain range,the pro-osteoclastic effect was positively correlated with the force conditions.At 45 kPa and 12 hours,RAW264.7 cells had the strongest osteoclastic differentiation effect.The osteoclast-promoting ability of 65 kPa and 12-hour group was weakened.(4)These findings indicated that the osteoclastic differentiation of RAW264.7 cells was promoted by three-dimensional cultured periodontal ligament stem cells under static compression through the paracrine pathway,and the force conditions were correlated with the osteoclast effect.

关 键 词：静压力 三维培养 牙周膜干细胞 RAW264.7细胞 破骨分化 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R856.78