Grzesik W J, Ivanov B, Robey F A, Southerland J, Yamauchi M
Department of Periodontics, Dental Research Center, School of Dentistry, University of North Carolina, Chapel Hill 27599-7455, USA.
J Dent Res. 1998 Aug;77(8):1606-12. doi: 10.1177/00220345980770080801.
Periodontal ligament (PDL) cells have been shown to express several integrins (alphav, alpha5, beta1, beta3) that use RGD (arginine-glycine-aspartic Acid)-dependent mechanisms for the recognition and binding of their ligands. The objective of this study was to evaluate the effects of certain integrin-binding cyclic and linear synthetic RGD-containing peptides on PDL cells' adhesion, proliferation, and de novo protein synthesis in vitro. Fifth passages of normal human PDL cells established from teeth extracted from patients (ages 12 to 14) for orthodontic reasons were used for all experiments. Synthetic peptides containing the EPRGDNYR sequence in two different spatial conformations (linear and cyclic) were covalently attached to bovine serum albumin (BSA). Type I collagen, EPRGDNYR-BSA conjugates, 1:1 mixtures of type I collagen and conjugates, as well as BSA (a negative control) were coated on bacteriological plastic and evaluated for their attachment-promoting activities. In addition, the effects of these substrates on cell proliferation were evaluated by [3H]thymidine incorporation by the PDL cells. For attachment and spreading, the cyclic forms of EPRGDNYR-BSA conjugate and type I collagen were most potent, followed by linear EPRGDNYR-BSA conjugate. The effects of all collagen/conjugate mixtures were equivalent to that of type I collagen except for the collagen/linear EPRGDNYR-BSA mixture, which was less potent. The cyclic EPRGDNYR-BSA conjugate was the most effective substrate to stimulate cell proliferation, and it was followed in potency by the linear peptide-BSA conjugate. Collagen alone did not stimulate [3H]thymidine incorporation above the control level. Mixtures of collagen with all of the conjugates showed stimulatory effects similar to that of the cyclic peptide-BSA conjugate. No significant differences in de novo protein synthesis were detected. These results suggest that the synthetic RGD-containing peptides attached to a carrier are potent ligands for the human PDL cells, and that they could provide a basis for the development of new strategies aimed at the regeneration of the periodontium.
牙周韧带（PDL）细胞已被证明可表达多种整合素（αv、α5、β1、β3），这些整合素利用依赖RGD（精氨酸-甘氨酸-天冬氨酸）的机制来识别和结合其配体。本研究的目的是评估某些整合素结合的环状和线性含RGD合成肽对PDL细胞体外黏附、增殖和从头蛋白质合成的影响。从因正畸原因拔除的患者（年龄12至14岁）牙齿中获取的正常人PDL细胞的第五代用于所有实验。将含有EPRGDNYR序列的两种不同空间构象（线性和环状）的合成肽共价连接到牛血清白蛋白（BSA）上。将I型胶原蛋白、EPRGDNYR-BSA缀合物、I型胶原蛋白与缀合物的1:1混合物以及BSA（阴性对照）包被在细菌学塑料上，并评估它们的促黏附活性。此外，通过PDL细胞掺入[3H]胸苷来评估这些底物对细胞增殖的影响。对于黏附和铺展，EPRGDNYR-BSA缀合物的环状形式和I型胶原蛋白最有效，其次是线性EPRGDNYR-BSA缀合物。除了胶原蛋白/线性EPRGDNYR-BSA混合物效果较差外，所有胶原蛋白/缀合物混合物的效果与I型胶原蛋白相当。环状EPRGDNYR-BSA缀合物是刺激细胞增殖最有效的底物，其次是线性肽-BSA缀合物。单独的胶原蛋白不会刺激[3H]胸苷掺入超过对照水平。胶原蛋白与所有缀合物的混合物显示出与环状肽-BSA缀合物类似的刺激作用。未检测到从头蛋白质合成有显著差异。这些结果表明，连接到载体上的含RGD合成肽是人类PDL细胞的有效配体，并且它们可为开发旨在牙周组织再生的新策略提供基础。
