不同来源成纤维细胞对角质形成细胞再上皮化影响  

Effects of fibroblasts derived from different sources on re-epithelialization of keratinocytes

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作  者:王丽[1] 潘乙怀[1] 邹多宏 Wang Li;Pan Yihuai;Zou Duohong(Institute of Stomatology,School and Hospital of Stomatology,Wenzhou Medical University,Wenzhou/Department of Endodontics,School and Hospital of Stomatology,Wenzhou Medical University,Wenzhou 325000;Shanghai Ninth People’s Hospital,School of Medicine,Shanghai Jiaotong University,Shanghai 200011)

机构地区:[1]温州医科大学口腔医学院附属口腔医院牙体牙髓病科,温州325000 [2]上海交通大学医学院附属第九人民医院,上海200011

出  处:《口腔材料器械杂志》2024年第3期150-157,共8页Chinese Journal of Dental Materials and Devices

基  金:国家自然科学基金(32171347);2022年度上海市卫生健康委员会卫生健康领军人才(2022XD038);上海交通大学医学院附属第九人民医院“临床研究助推计划”(JYLJ202102)。

摘  要:目的比较人牙龈及皮肤来源成纤维细胞条件培养基对永生化角质形成细胞再上皮化作用的差异,分析成纤维细胞促进再上皮化作用的关键因子,探究牙龈来源成纤维细胞在创伤修复中的可能应用。方法分别制备人牙龈成纤维细胞(hGFs)和人真皮成纤维细胞(hDFs)条件培养基(CM),添加到人永生化角质形成细胞(Human keratinocyte cells,HaCaT)中,采用CCK-8、EdU染色检测HaCaT细胞增殖能力;细胞划痕实验、Transwell细胞迁移实验检测细胞迁移能力;通过ABplex多因子检测及酶联免疫吸附法(ELISA)对hGFs、hDFs分泌的多种与创伤修复相关的生长因子、细胞因子和趋化因子进行定量分析。结果CCK-8、EdU染色、细胞划痕实验、Transwell细胞迁移实验结果表明:2种条件培养基均能显著提高HaCaT细胞的增殖及迁移能力(P<0.05),hGFs-CM对HaCaT细胞增殖和迁移能力的促进作用显著强于hDFs-CM(P<0.01);hGFs和hDFs分泌的因子有较大的差异,其中hGFs分泌的生长因子如肝细胞生长因子(HGF)、血管内皮生长因子(VEGF)明显多于hDFs。结论人牙龈来源成纤维细胞较人皮肤来源成纤维细胞的条件培养基对角质形成细胞的增殖和迁移能力的有更强的促进作用,可能与其旁分泌产生的因子不同有关。Objective This study aimed to compare the effects of conditioned medium derived from human gingival fibroblasts(hGFs)and dermal fibroblasts(hDFs)on the re-epithelialization of immortalized keratinocytes(HaCaT cells),explore the critical factors involved in the role of fibroblasts in promoting re-epithelialization,and analyze the possible application of gingival-derived fibroblasts in wound healing.Methods The conditioned medium(CM)of hGFs and hDFs were prepared and used to incubate with HaCaT cells,respectively.The proliferation ability of HaCaT cells was detected by CCK-8 and EdU staining;the cell migration ability was detected by cell scratch assay and Transwell cell migration assay;and a variety of growth factors,cytokines,and chemokines related to wound healing secreted by hGFs and hDFs were quantitatively analyzed by the ABplex custom panel assay kit and enzyme-linked immunosorbent assay(ELISA).Results The results of CCK-8,EdU staining,cell scratch assay,and Transwell cell migration assay demonstrated a significant enhancement in the proliferation and migration ability of HaCaT cells upon treatment with hGFs-CM and hDFs-CM(P<0.05).Notably,hGFs-CM exhibited a more pronounced effect on promoting HaCaT cell proliferation and migration than hDFs-CM(P<0.01).There were significant differences in the secreted factors between hGFs and hDFs,with hGFs secreting significantly more growth factors such as hepatocyte growth factor(HGF)and vascular endothelial growth factor(VEGF)than hDFs.Conclusion The proliferation and migratory ability of keratinocytes were more strongly promoted by the conditioned medium of human gingival-derived fibroblasts than that of human skin-derived fibroblasts,which may be related to differences in the factors produced paracrine.

关 键 词:创伤修复 牙龈成纤维细胞 真皮成纤维细胞 永生化角质形成细胞 再上皮化 

分 类 号:R78[医药卫生—口腔医学]

 

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