Production of Transforming Growth Factor-beta1 in Human Fibroblasts Induced with Bacterial Toxins

Lee, Seong Geun; Kim, Kwang Hyuk

Abstract

TGF-beta1 is a potent chemotactic factor for inflammatory cells and fibroblasts. It also stimulates the celluar source and components of extracellular matrix and the production of proteinase inhibitors. Collectively, these biologic activities lead to the accumulation and stabilization of the nascent matrix, which is vital to wound healing. The objective of this study is to investigate production of TGF-beta1 in vitro fibroblast culture in the presence of Staphylococcus enterotoxin B(SEB) and/or lipopolysaccharide(LPS) and to elucidate the role of TGF-beta1 which may be responsible for wound healing The fibroblasts were originated from facial dermis and hypertrophic scar in 26 year-old male patient. In the presence of LPS(0.0l microgram, 0.1 microgram, 1.0 microgram), SEB(0.0l microgram, 0.1 microgram, 1.0 microgram) respectively, cells(5x103ml) were cultivated in vitro. At 1, 3, and 5 days after incubation, cells were counted. Also, cells(2.5x105ml) were cultivated in EMEM with LPS(0.01, 0.1 and 1.0 microgram), SEB(0.01, 0.1 and 1.0 microgram) respectively and LPS(0.1 microgram) and SEB(0.1 microgram) in combination for 24, 48, and 72 hours respectively. Culture supernatants were harvested at 1, 2, and 3 days after incubation period and triplicate culture supernatants were pooled and TGF-beta1 was assayed in duplicate. The results were as follows. 1. In facial dermal fibroblast induced with SEB and LPS respectively or in combination, the suppression of cell proliferation occurred very significantly at 1 day after incubation, compared with the control. In SEB exposure, the production of TGF-beta1 was decreased very significantly at 1 day after incubation, compared with the control. However, in LPS, SEB and LPS exposure, the production of TGF-beta1 was increased very significantly at 1 day after incubation, compared with the control. 2. In hypertrophic scar fibroblast induced with SEB and LPS respectively or in combination, the suppression of cell proliferation did not occur at 1 day after incubation, compared with the control. In SEB and LPS exposure in combination, the production of TGF-beta1 was increased very significantly at 1 day after incubation, compared with the control. However, the production of TGF-beta1 did not occur in SEB and LPS exposure respectively. In conclusion, the concentration of bacterial toxins and the incubation period correlated with cell proliferation and production of TGF-beta1 very significantly and both fibroblasts have different phenotype each other in this regard. This data suggest that the significant production of TGF-beta1 may develope abnormal wound healing associated with tissue fibroproliferative disorder, such as hypertrophic scar and keloid formation.