Mailhot J M, Schuster G S, Garnick J J, Hanes P J, Lapp C A, Lewis J B
Department of Periodontics, Medical College of Georgia, Augusta 30912, USA.
J Clin Periodontol. 1995 Sep;22(9):679-85. doi: 10.1111/j.1600-051x.1995.tb00826.x.
The purpose of this study was to measure the time-sequence response of RNA and protein synthesis to transforming growth factor-beta 1 (TGF-beta 1) by human periodontal ligament (HPDLF) and gingival (HGF) fibroblasts in culture. HPDLF and HGF were cultured from explants of healthy gingival tissue and freshly extracted teeth. Cultures of 8 x 10(4) cells/ml were exposed to medium containing 3H-uridine and 35S-methionine with TGF-beta 1 at concentrations from 10(-9) M to 10(-21) M, or control medium, for up to 60 hours in order to assess RNA and protein synthesis. Protein concentrations of comparable cultures were also assayed colorimetrically. Results were reported as specific activity (CPM/microgram protein). The results indicate that 10(-9) M TGF-beta 1 treated cultures showed a significant increase in RNA synthesis by HPDLF and HGF over time, as compared to the control cultures. HPDLF showed a significant increase in protein synthesis over time while that by HGF was not significant as compared to the control cultures. Lower concentrations of TGF-beta 1 demonstrated no significant differences from control. Results suggest that the effects of TGF-beta 1 on HPDLF and HGF are both time and dose dependent, with 10(-9) M TGF-beta 1 providing the best response of those concentrations tested. These findings support the concept that TGF-beta 1 may play a role in periodontal regeneration due to its ability to promote fibroblast RNA and protein synthesis. The results also demonstrate that although these two cells types appear morphologically similar, they exhibit distinct biological responses to growth factors such as TGF-beta 1.
本研究的目的是测定培养的人牙周膜成纤维细胞(HPDLF)和牙龈成纤维细胞(HGF)中RNA和蛋白质合成对转化生长因子-β1(TGF-β1)的时间序列反应。HPDLF和HGF从健康牙龈组织和新鲜拔除牙齿的外植体中培养获得。将8×10⁴个细胞/ml的培养物暴露于含有³H-尿苷和³⁵S-甲硫氨酸以及浓度从10⁻⁹M至10⁻²¹M的TGF-β1的培养基或对照培养基中,长达60小时,以评估RNA和蛋白质合成。还通过比色法测定了可比培养物的蛋白质浓度。结果以比活性(CPM/微克蛋白质)报告。结果表明,与对照培养物相比,10⁻⁹M TGF-β1处理的培养物中HPDLF和HGF的RNA合成随时间显著增加。与对照培养物相比,HPDLF的蛋白质合成随时间显著增加,而HGF的蛋白质合成则不显著。较低浓度的TGF-β1与对照无显著差异。结果表明,TGF-β1对HPDLF和HGF的作用具有时间和剂量依赖性,在所测试的浓度中,10⁻⁹M TGF-β1产生的反应最佳。这些发现支持了TGF-β1可能因其促进成纤维细胞RNA和蛋白质合成的能力而在牙周再生中发挥作用的概念。结果还表明,尽管这两种细胞类型在形态上相似,但它们对诸如TGF-β1等生长因子表现出不同的生物学反应。