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目的:通过检测米诺环素对体外培养的人牙周膜成纤维细胞(hPDLF)增殖、蛋白合成、Ⅰ型胶原分泌、碱性磷酸酶活性、骨钙素水平的影响,探讨米诺环素局部给药的效应浓度范围。方法:将不同浓度的米诺环素(0、10、20、40、100、200mg/L)分别作用于第5代(P5代)hPDLF,孵育1、4、7、10、14d后,采用MTT法检测其对细胞增殖的影响,以及对细胞碱性磷酸酶活性、蛋白质合成、Ⅰ型胶原分泌、骨钙素分泌的影响。实验结果采用SPSS13.0软件包进行分析。结果:10～200mg/L米诺环素显著促进P5代hPDLF的增殖活性(P<0.05),100mg/L为最大效应浓度;20～200mg/L明显促进P5代hPDLF蛋白质合成及Ⅰ型胶原分泌(P<0.05);10mg/L显著促进碱性磷酸酶活性(P<0.05),20～200mg/L显著抑制碱性磷酸酶活性(P<0.05);40～200mg/L显著抑制骨钙素分泌(P<0.05)。结论:在10～20mg/L的浓度范围内,米诺环素刺激P5代hPDLF细胞增殖,促进细胞分化,促进蛋白质合成,促进Ⅰ型胶原分泌,对hPDLF分泌骨钙素无抑制作用。10～20mg/L可作为米诺环素局部给药的效应浓度范围。
OBJECTIVE: To investigate the effects of minocycline on proliferation, protein synthesis, type I collagen secretion, alkaline phosphatase activity and osteocalcin levels in cultured human periodontal ligament fibroblasts (hPDLF) The range of effect concentrations for topical administration. Methods: Different concentrations of minocycline (0, 10, 20, 40, 100 and 200 mg / L) were applied to the fifth generation (P5 generation) hPDLF respectively and incubated for 1, 4, 7, MTT assay was used to detect the effect on cell proliferation, cell alkaline phosphatase activity, protein synthesis, type I collagen secretion and osteocalcin secretion. Experimental results using SPSS13.0 software package for analysis. Results: Minocycline at 10 ~ 200mg / L significantly promoted the proliferation of hPDLF at P5 generation (P <0.05), and 100mg / L at maximum effect concentration. 20 ~ 200mg / L significantly promoted hPDLF protein synthesis and type Ⅰ collagen secretion (P <0.05); alkaline phosphatase (P <0.05) was significantly enhanced at 10 mg / L and alkaline phosphatase (P <0.05) at 20 ~ 200 mg / Secretion (P <0.05). CONCLUSION: Minocycline stimulates the proliferation of P5 generation hPDLF cells at 10 ~ 20 mg / L, promotes cell differentiation, promotes protein synthesis, and promotes the secretion of type I collagen. It has no inhibitory effect on the secretion of osteocalcin by hPDLF. 10 ~ 20mg / L minocycline can be used as local drug concentration range.