The study of collagen synthesis of human periodontal fibroblasts

为了解牙周病相关细菌的脂多糖对人牙周成纤维细胞的影响，采用正丁醇萃取法从牙龈拟杆菌381和中间拟杆菌ATCC 25611中提取脂多糖。使用来自大肠杆菌Olll：B_4（List Biological Laboratory）的脂多糖。从人健康牙龈、人发炎牙龈和人牙周韧带中获得成纤维细胞，并在细胞培养基中生长。从健康受试者的血液中收集单核细胞。将几种细菌的脂多糖和^3H-脯氨酸加入成纤维细胞的培养基中。放射性测定成纤维细胞合成胶原。将脂多糖添加到单核细胞的培养基中，孵育24小时后，收集培养基上清。将此上清液加入成纤维细胞培养液中，测定胶原合成。单核细胞培养上清和脂多糖均能抑制其摄取^3H-脯氨酸的能力。牙周膜成纤维细胞对^3H-脯氨酸摄取能力最低。脂多糖和单核细胞培养上清均能抑制3种成纤维细胞的胶原合成能力，但在成纤维细胞培养上清中加入不含脂多糖的单核细胞培养上清后，这种能力增强。加入脂多糖对胶原合成与蛋白掺入的比例无明显影响，加入单核细胞培养液上清可降低胶原合成与蛋白掺入的比例，其中牙周膜成纤维细胞胶原合成与蛋白掺入的比例最高。

In order to know the influence of the lipopolysaccharides from periodontal disease related bacteria to the human periodontal fibroblasts, this study was performed.Lipopolysaccharides were extracted from Bacteroides gingivalis 381 and Bacteroides intermedius ATCC 25611 by butanol extract method. Lipopolysaccharide from Escherichia Coli Olll:B_4 (List Biological Laboratory) was used. Fibroblasts were obtained from human healthy gingiva, from human inflamed gingival and from human periodontal ligament with outgrowth in the cell culture medium. Monocyte were collected from the blood of a healthy subject. Lipopolysaccharides from several bacteria and ^3H-proline were added into the culture medium of fibroblasts. Synthesized collagen by fibroblasts were measured radio-actively. Lipopolysaccharides were also added into the culture medium of monocute and after 24 hr incubation, supernant of culture medium was collected. This supernant ^3H-proline were added into the culture medium of fibroblasts and synthesized collagen was measured. The intake ability of ^3H-proline into the protein was depressed by lipopolysaccharides and by the supernant of monodyte culture medium. Periodontal ligament fibroblast showed the most low intake ability of ^3H-proline into-protein. The collagen synthesis ability of 3 types of fibroblasts were depressed by lipopolysaccharides and by the supernant of monocyte culture medium.But when supernant of monocyte culture medium without lipopolysaccharides was added into the medium of fibroblasts, this ability was increased. The ratio of collagen synthesis to protein corporation was not changed when lipopolysaccharides were added.This ratio was decreased by the addition of supernant of monocute culture medium.The highest ratio was observed in the fibroblast from periodontal ligament.