The study of collagen synthesis of human periodontal fibroblasts

为了解牙周病相关菌脂多糖对人牙周成纤维细胞的影响，采用正丁醇提取法从牙周类杆菌381和中间类杆菌ATCC 25611中提取脂多糖。使用大肠杆菌Oll：B4(LIST生物实验室)的脂多糖。成纤维细胞取自人的健康牙龈，取自人发炎的牙龈，取自人的牙周膜，在细胞培养液中生长。单核细胞是从健康受试者的血液中采集的。在成纤维细胞的培养液中加入几种细菌的脂多糖和~3H-脯氨酸。放射活性测定成纤维细胞合成的胶原蛋白。在单细胞培养上清液中加入脂多糖，培养24小时后收集上清液。将此上清液加入成纤维细胞的培养液中，测定胶原蛋白的合成。内毒素和单倍体培养上清液均可抑制~3H-Pro的摄取能力。牙周膜成纤维细胞对~3H-脯氨酸摄取能力最低。内毒素和单核细胞培养上清液对3种成纤维细胞的胶原合成能力均有抑制作用，但不含脂多糖的单核细胞培养上清液对成纤维细胞的胶原合成能力有促进作用。加入脂多糖后，胶原合成与蛋白质结合的比例无明显变化，加入单次培养上清液后，胶原合成与蛋白结合的比例降低，牙周膜成纤维细胞的胶原合成与蛋白质合成比例最高。

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.