苯妥英钠对人牙周膜成纤维细胞生物学活性影响的实验研究  

Biological effects of phenytoin on cultured human periodontal ligament fibroblasts in vitro

作　　者：于美娇[1] 杨丕山[1] 葛少华[1] 

机构地区：[1]山东大学口腔医院牙周病科,山东济南250012

出　　处：《华西口腔医学杂志》2008年第2期215-218,共4页West China Journal of Stomatology

基　　金：国家自然科学基金资助项目(30371537);山东省自然科学基金资助项目(21350000413011)

摘　　要：目的研究苯妥英钠(PHT)对体外培养的人牙周膜成纤维细胞(hPDLF)生物学功能的影响,并探讨其用于促进牙周组织再生的可能性。方法将不同质量浓度的PHT(1、5、20、100、500、2500mg/L)加入体外培养的第4代hPDLF,检测其对细胞增殖活性、蛋白合成、碱性磷酸酶(ALP)活性的影响;Von Kossa染色法检测PHT(20、100、500mg/L)对第4代hPDLF矿化结节形成能力的影响;应用ELISA法测定PHT(20、100mg/L)对第3代hPDLF中骨形态发生蛋白-2(BMP-2)表达的影响。结果在20、100mg/L的质量浓度时,PHT可显著促进hPDLF的增殖及分化(P<0.01);在质量浓度100mg/L时,PHT可增强hPDLF的蛋白合成(P<0.05);同时,PHT在质量浓度100mg/L可显著促进hPDLF的矿化能力及BMP-2的表达(P<0.01)。但高质量浓度(2500mg/L)的PHT则严重抑制细胞的生物学活性。结论适当质量浓度的PHT对hPDLF的增殖和分化有促进作用,但过高质量浓度的PHT具有细胞毒性,不利于牙周组织的修复和再生。Objective To study the biological effects of phenytoin （PHT） on cultured human periodontal ligament fibroblasts （hPDLF）, and explore the possibility of its accelerating periodontal regeneration. Methods Increasing concentrations of PHT （1, 5, 20, 100, 500, 2 500 mg/L） were added into the medium of the fourth passage of cultured hPDLF, respectively. After co-incubated for 3 days, cell proliferation activity, the total amount of protein and alkaline phosphatase（ALP） activity were detected. Mineralized sodium and PHT（20, 100, 500 mg/L） were added into the medium of the fourth passage hPDLF. After co-incubated, the mineralized nodules formation were detected by Von Kossa staining. The third passage hPDLF were stimulated by PHT （20, 100 mg/L）, bone morphogenetic protein-2（BMP-2） concentration was analyzed by enzyme linked immunosorbent sandwich assay （ELISA）. Results At the concentration of 20 or 100 mg/L, PHT significantly enhanced the proliferating activity and ALP activity of hPDLF （P〈0.01）. PHT at 100 mg/L could increase protein synthesis of hPDLF（P〈0.05）. The capability of mineralization and BMP-2 expression of hPDLF were increased significantly （P〈0.01） in 100 mg/L group when compared with that in the control group. However, higher concentration（2 500 rag/L） not only changed cell morphology, but also significantly inhibited cell activity. Conclusion The results suggested that proper doses of PHT could promote proliferation and biosynthesis and also enhance osteogenesis by increasing the differentiation, mineralization and BMP-2 expression of hPDLF while higher concentrations of PHT had cytotoxic effect.

关 键 词：苯妥英钠 牙周膜成纤维细胞 细胞培养 矿化 

分 类 号：R78[医药卫生—口腔医学]