摘要
目的:探究骨髓巨噬细胞主穹窿蛋白(MVP)对骨再生修复的影响。方法:在野生型(WT)C57BL/6小鼠和特异性敲除巨噬细胞MVP(CKO)的小鼠中分别构建胫骨骨皮质缺损模型,通过Micro⁃CT扫描与重建技术、苏木素⁃伊红染色观察骨再生情况;体外分离培养WT和CKO小鼠骨髓来源的巨噬细胞,并分别与野生型骨髓间充质干细胞(BMSC)共培养,通过碱性磷酸酶染色、成骨分化标记基因实时荧光定量PCR实验,观察敲除巨噬细胞MVP对共培养体系中的BMSC成骨向分化的调控作用;通过细胞因子芯片实验检测巨噬细胞细胞因子的分泌。结果:骨缺损第14天,Micro⁃CT扫描及重建结果显示CKO小鼠缺损区矿化组织少于WT小鼠(P<0.05);苏木素⁃伊红染色结果显示CKO组小鼠缺损区骨痂面积小于WT小鼠;与CKO组巨噬细胞共培养的BMSC的成骨分化标记基因水平显著低于与WT组巨噬细胞共培养组(P<0.05);敲除MVP的巨噬细胞分泌更多抑制成骨的细胞因子CCL2、CCL4、CXCL10、CCL3、IL⁃7。结论:巨噬细胞MVP可能通过促进BMSC成骨分化进而调控骨再生修复。
Objective:To investigate the role of macrophage major vault protein(MVP)on bone regeneration.Methods:Macro⁃phage⁃specific MVP knockout(CKO)and wild⁃type(WT)C57BL/6 mice were used to establish a tibial mono⁃cortical defect model,and the bone regeneration was observed by Micro⁃CT and hematoxylin⁃eosin(HE)staining.Macrophages from WT or CKO mice bone marrow were co⁃cultured with wild⁃type bone marrow mesenchymal stem cell(BMSC),respectively.The effect of macrophage MVP on BMSC osteogenic differentiation was tested by alkaline phosphatase(ALP)staining and qRT⁃PCR.Finally,a cytokine array was used to test the cytokine profile of macrophages to understand the regulation of MVP on cytokines secretion.Results:On day 14 of bone de⁃fect,Micro⁃CT scan and reconstruction results showed that the mineralized tissue in the defect area of CKO mice was less than that of WT mice(P<0.05);hematoxylin⁃eosin staining results showed that the bone area in the defect area of CKO mice was less than that of WT mice;In the in vitro co⁃culture assay,the levels of osteogenic differentiation marker genes were significantly lower in BMSC co⁃cul⁃tured with CKO macrophages than in those co⁃cultured with WT macrophages(P<0.05).Cytokine array results showed that macropha⁃ges lacking MVP secreted more cytokines including CCL2,CCL4,CXCL10,CCL3,IL⁃7 that inhibit osteogenic differentiation.Con⁃clusions:Macrophage MVP may regulate bone regeneration by promoting BMSC osteogenic differentiation.
作者
杨燕
赵娜
马俊青
YANG Yan;ZHAO Na;MA Junqing(Department of Orthodontics,The Affiliated Stomatological Hospital of Nanjing Medical University,Jiangsu Province Key Laboratory of Oral Diseases,Jiangsu Province Engineering Research Center of Stomatological Translational Medi-cine,Nanjing 210029,China)
出处
《口腔生物医学》
2023年第3期151-155,共5页
Oral Biomedicine
基金
国家自然科学基金(82170911)
江苏省科教能力提升工程——江苏省研究型医院(YJXYYJSDW4)
江苏省医学创新中心(CXZX202227)。
关键词
主穹窿蛋白
巨噬细胞
骨再生
成骨分化
细胞因子
major vault protein
macrophage
bone regeneration
osteogenic differentiation
cytokine
