Kent L W, Dyken R A, Rahemtulla F, Allison A C, Michalek S M
Department of Microbiology, School of Dentistry, University of Alabama at Birmingham 35294-2170, USA.
Arch Oral Biol. 1996 Mar;41(3):263-70. doi: 10.1016/0003-9969(95)00127-1.
Cytokines have been implicated in the regulation of antibody and inflammatory responses, but their role in periodontal diseases has not been elucidated. In the present study, cytokine production by human gingival fibroblasts (HGF) following in vitro passage was assessed in order to determine the basal levels of cytokine message and protein and to determine if the cellular morphology and the profile of cytokines produced differed with passage. The HGF cell line F-CL was established by explantation from non-inflamed gingival tissue, and cells from passages 1-10 were studied. The number of cells was determined in confluent cultures and cell morphology was examined by light microscopy. Fibroblasts from confluent cultures were examined for cytokine mRNA by reverse transcription-polymerase chain reaction and culture supernatants were assessed for cytokines by enzyme-linked immunosorbent assay. The morphology of F-CL fibroblasts in passages 1-4 was normal, while fibroblasts in passages 5-10 were larger. In general, the number of cells decreased from early to late passage. Fibroblasts from passages 1-10 contained message for interleukin-1 beta, -6 and -8, but not for interleukin-1 alpha or tumour necrosis factor-alpha. Interleukin-6 was detected in culture supernatants of F-CL fibroblasts by the enzyme immunoassay and its level decreased with increasing passage.
细胞因子与抗体及炎症反应的调节有关，但其在牙周疾病中的作用尚未阐明。在本研究中，评估了体外传代后人牙龈成纤维细胞（HGF）的细胞因子产生情况，以确定细胞因子信息和蛋白质的基础水平，并确定细胞形态和产生的细胞因子谱是否随传代而不同。通过从非炎症性牙龈组织外植建立了HGF细胞系F-CL，并研究了第1 - 10代的细胞。在汇合培养物中确定细胞数量，并通过光学显微镜检查细胞形态。通过逆转录 - 聚合酶链反应检测汇合培养物中的成纤维细胞的细胞因子mRNA，并通过酶联免疫吸附测定评估培养上清液中的细胞因子。第1 - 4代F-CL成纤维细胞的形态正常，而第5 - 10代的成纤维细胞更大。一般来说，细胞数量从传代早期到晚期减少。第1 - 10代的成纤维细胞含有白细胞介素 - 1β、 - 6和 - 8的信息，但不含有白细胞介素 - 1α或肿瘤坏死因子 - α的信息。通过酶免疫测定在F-CL成纤维细胞的培养上清液中检测到白细胞介素 - 6，其水平随传代增加而降低。
