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目的:通过血管内皮细胞与牙周组织细胞的复合培养模拟牙周组织再生环境,研究血管内皮细胞对牙周组织细胞迁移的影响,为明确血管内皮细胞在牙周组织再生过程中的作用奠定基础。方法:应用原代培养的血管内皮细胞,在Transwell嵌套中实现与人牙周膜成纤维细胞、牙龈成纤维细胞间的复合培养。分别以Transwell嵌套滤膜下腔面24 h的细胞数量检测垂直迁移能力,以划痕实验后0、8、16和24 h计数划痕区域的细胞数量检测水平迁移能力.以玻片试验及计算机辅助图像处理软件计算实验第1、4和7天细胞覆盖创面的面积检测覆盖创面能力.并均与单独培养的2种牙周组织细胞作为对照。采用SPSS13.0软件包对实验结果进行统计学分析。结果:血管内皮细胞存在时,2种牙周细胞在垂直和水平方向的迁移量均显著高于单独培养组(P<0.01),且牙周膜成纤维细胞垂直向的迁移量显著高于牙龈成纤维细胞(P<0.01),但其水平迁移量在实验24 h时则显著低于牙龈成纤维细胞(P<0.01):在血管内皮细胞共存条件下,2种牙周细胞创面覆盖能力在实验第7天高于对照组,且牙周膜成纤维细胞的创面覆盖能力的增加显著高于牙龈成纤维细胞(P<0.01)。结论:血管内皮细胞对牙周膜、牙龈成纤维细胞的迁移和创面覆盖有明显促进作用,且血管内皮细胞对牙周膜成纤维细胞的垂直向迁移和创面覆盖能力的促进作用更为明显。
OBJECTIVE: To study the effects of vascular endothelial cells on the migration of periodontal tissues through the complex culture of vascular endothelial cells and periodontal tissues, and to establish the basis for clarifying the role of vascular endothelial cells in periodontal tissue regeneration . Methods: The cultured primary cultured vascular endothelial cells were cultured in transwell nested with human periodontal ligament fibroblasts and gingival fibroblasts. The vertical migrating capacity was measured by the number of cells in the transwell nesting subfascial surface for 24 h, respectively, and the horizontal migration ability was detected by counting the number of cells in the scratch area at 0, 8, 16 and 24 h after scratch test. Computer-aided image processing software was used to calculate the area covered by wounds covered by cells on days 1, 4 and 7, and the wound healing ability was compared with that of 2 periodontal tissues separately cultured. SPSS13.0 software package was used to analyze the experimental results. Results: In the presence of vascular endothelial cells, the migration of the two periodontal cells in both vertical and horizontal directions was significantly higher than that of the cultured group (P <0.01), and the vertical migration of periodontal ligament fibroblasts was significantly higher than that of the gums Fibroblasts (P <0.01), but the level of migration was significantly lower than that of gingival fibroblasts at 24 h (P <0.01). The wound healing ability of two periodontal cells in the presence of vascular endothelial cells was On the 7th day of experiment, the wound coverage of periodontal ligament fibroblasts was significantly higher than that of the control group (P <0.01). CONCLUSION: Vascular endothelial cells can significantly promote the migration and wound coverage of periodontal ligament and gingival fibroblasts, and the effect of vascular endothelial cells on the vertical migration and wound healing of periodontal ligament fibroblasts is more obvious.