Gago L April, Saed Ghassan M, Wang Rona X, Kruger Michael, Diamond Michael P
Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Wayne State University School of Medicine, 4707 St. Antoine Boulevard, Detroit, MI 48201, USA.
Am J Obstet Gynecol. 2003 Dec;189(6):1620-5; discussion 1625-6. doi: 10.1016/j.ajog.2003.08.018.
The purpose of this study was to evaluate the potential biologic effects of oxidized regenerated cellulose, which has been shown in multiple human in vivo studies to reduce postoperative adhesion development, on the messenger RNA levels of transforming growth factor-beta1, type I collagen, type III collagen, and fibronectin.
The oxidized regenerated cellulose was dissolved in saline solution and added to confluent, monolayer cultures of human normal fibroblasts and mesothelial cells. Control cells were maintained in media alone at the same pH. After 24 hours of treatment, total RNA was extracted from all cells. Real-time reverse transcription-polymerase chain reaction was performed to determine the relative change in messenger RNA levels of type I, type HI collagen, fibronectin, transforming growth factor-beta1, and beta-actin (housekeeping gene) in response to the oxidized regenerated cellulose treatment (n=4 cultures). Student t tests were performed for each cell type, which compared oxidized regenerated cellulose-treated cells to control cells. Calculated power for the statistically significant findings ranged from 65% to 100%.
Transforming growth factor-beta1 messenger RNA was elevated by the oxidized regenerated cellulose treatment in the mesothelial cells by 13% (control cells, 0.562+/-0.022; oxidized regenerated cellulose-treated cells, 0.636+/-0.014; P=.03). In normal fibroblasts, transforming growth factor-beta1 messenger RNA was slightly, but not significantly, decreased in oxidized regenerated cellulose-exposed normal fibroblasts compared with controls (control cells, 0.622+/-0.062; oxidized regenerated cellulose-treated cells, 0.609+/-0.006; P=.85). Type I collagen was found to be increased by exposure to oxidized regenerated cellulose in both mesothelial cells and normal peritoneal fibroblasts. Type I collagen was increased by 23% in mesothelial cells (control cells [0.587+/-0.018] vs oxidized regenerated cellulose-treated cells [0.722+/-0.010], P=.002), and 27% in normal fibroblasts (control cells, 0.522+/-0.018, oxidized regenerated cellulose-treated cells, 0.665+/-0.009; P=.001). However, messenger RNA levels of type III collagen and fibronectin (other representative molecules of the extracellular matrix) were not altered significantly by oxidized regenerated cellulose application in vitro.
Oxidized regenerated cellulose influences the expression of factors that are accepted commonly to be associated with adhesiogenesis. Oxidized regenerated cellulose increased the expression of transforming growth factor-beta1 in mesothelial cells and type I collagen in mesothelial cells and normal peritoneal fibroblasts. Minimization of these oxidized regenerated cellulose-induced molecular changes, if they occur in vivo, may improve the ability of oxidized regenerated cellulose to reduce postoperative adhesion development.
本研究旨在评估氧化再生纤维素对转化生长因子-β1、I型胶原、III型胶原和纤连蛋白信使核糖核酸水平的潜在生物学效应，氧化再生纤维素已在多项人体体内研究中显示可减少术后粘连形成。
将氧化再生纤维素溶解于盐溶液中，并添加到汇合的人正常成纤维细胞和间皮细胞单层培养物中。对照细胞仅在相同pH值的培养基中培养。处理24小时后，从所有细胞中提取总RNA。进行实时逆转录-聚合酶链反应，以确定I型、III型胶原、纤连蛋白、转化生长因子-β1和β-肌动蛋白（管家基因）信使核糖核酸水平在氧化再生纤维素处理后的相对变化（n = 4个培养物）。对每种细胞类型进行学生t检验，将氧化再生纤维素处理的细胞与对照细胞进行比较。统计显著结果的计算功效范围为65%至100%。
氧化再生纤维素处理使间皮细胞中转化生长因子-β1信使核糖核酸升高13%（对照细胞，0.562±0.022；氧化再生纤维素处理的细胞，0.636±0.014；P = 0.03）。在正常成纤维细胞中，与对照相比，暴露于氧化再生纤维素的正常成纤维细胞中转化生长因子-β1信使核糖核酸略有下降，但不显著（对照细胞，0.622±0.062；氧化再生纤维素处理的细胞，0.609±0.006；P = 0.85）。发现暴露于氧化再生纤维素后，间皮细胞和正常腹膜成纤维细胞中的I型胶原均增加。间皮细胞中I型胶原增加23%（对照细胞[0.587±0.018]与氧化再生纤维素处理的细胞[0.722±0.010]，P = 0.002），正常成纤维细胞中增加27%（对照细胞，0.522±0.018，氧化再生纤维素处理的细胞，0.665±0.009；P = 0.001）。然而，体外应用氧化再生纤维素并未显著改变III型胶原和纤连蛋白（细胞外基质的其他代表性分子）的信使核糖核酸水平。
氧化再生纤维素影响通常被认为与粘连形成相关的因子的表达。氧化再生纤维素增加了间皮细胞中转化生长因子-β1的表达以及间皮细胞和正常腹膜成纤维细胞中I型胶原的表达。如果这些氧化再生纤维素诱导的分子变化发生在体内，将其最小化可能会提高氧化再生纤维素减少术后粘连形成的能力。
