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Both transforming growth factor-beta (TGF-beta) and platelet-derived growth factor (PDGF) have been shown to affect cell proliferation in vitro. The hypothesis being tested was that the effects of the 2 cytokines would be modulated by the presence of serum in the medium. Gingival fibroblasts, obtained from periodontally healthy patients, were maintained in primary culture. Dose response experiments were performed for each growth factor in serum-free medium and in medium containing natural or heat-inactivated fetal bovine serum (10% FBS). Changes in cell numbers were quantified by crystal violet staining. The optimal concentrations of the individual factors (10 ng/ml TGF-beta1, 20 ng/ ml PDGF-BB) were then used when the 2 factors were tested in various sequences. In serum-free medium or in medium with 10% natural serum, the response to PDGF-BB was dose-dependent up to 40 ng/ml; however, with 10% heat-inactivated serum, the maximal response was seen at 20 ng/ml. The largest increase in cell numbers was produced by the simultaneous exposure to the two cytokines, rather than a sequential presentation. The findings suggest that the 48-h growth response of human gingival fibroblasts to 10 ng/ml TGF-beta1 or 20 ng/ ml PDGF-BB in serum-free medium was equivalent to growth obtained in medium containing heat-inactivated 10% FBS without added growth factors.

The purpose of this study was to measure the time-sequence response of RNA and protein synthesis to transforming growth factor-beta 1 (TGF-beta 1) by human periodontal ligament (HPDLF) and gingival (HGF) fibroblasts in culture. HPDLF and HGF were cultured from explants of healthy gingival tissue and freshly extracted teeth. Cultures of 8 x 10(4) cells/ml were exposed to medium containing 3H-uridine and 35S-methionine with TGF-beta 1 at concentrations from 10(-9) M to 10(-21) M, or control medium, for up to 60 hours in order to assess RNA and protein synthesis. Protein concentrations of comparable cultures were also assayed colorimetrically. Results were reported as specific activity (CPM/microgram protein). The results indicate that 10(-9) M TGF-beta 1 treated cultures showed a significant increase in RNA synthesis by HPDLF and HGF over time, as compared to the control cultures. HPDLF showed a significant increase in protein synthesis over time while that by HGF was not significant as compared to the control cultures. Lower concentrations of TGF-beta 1 demonstrated no significant differences from control. Results suggest that the effects of TGF-beta 1 on HPDLF and HGF are both time and dose dependent, with 10(-9) M TGF-beta 1 providing the best response of those concentrations tested. These findings support the concept that TGF-beta 1 may play a role in periodontal regeneration due to its ability to promote fibroblast RNA and protein synthesis. The results also demonstrate that although these two cells types appear morphologically similar, they exhibit distinct biological responses to growth factors such as TGF-beta 1.

Purpose: This study compared the relative affinity of Porphyromonas gingivalis and Escherichia coli endotoxin, bacterial cell envelope lipopolysaccharide (LPS), for three provisional resins., Materials and Methods: As-polymerized and pumiced polymethyl methacrylate and polyethyl methacrylate resin discs were exposed to 1,000 endotoxin U/mL P. gingivalis or E. coli LPS in water for 24 hours at 37 degrees C, whereas control discs were placed in LPS-free water. LPS-treated discs were transferred at 24-hour intervals to fresh, LPS-free water for up to 96 hours, and the incubated eluates were tested for the presence of LPS., Results: Initial adherence of P. gingivalis LPS to as-polymerized and pumiced-finish resin was a function of resin type, but surface characteristics modified adherence levels. When steady rates of elution were reached at 72 to 96 hours, as-polymerized specimens released significantly greater LPS levels than pumiced samples. Comparison of initial adherence of P. gingivalis and E. coli LPS with pumiced resins showed that adherence was based on a combination of LPS and resin type. P. gingivalis LPS had a greater relative affinity for polyethyl methacrylate, and E. coli LPS has a greater relative affinity for polymethyl methacrylate. Regardless of resin type, P. gingivalis LPS eluted at levels greater than E. coli LPS., Conclusions: The affinity of LPS for provisional resins seems to be a function of selective interactions based on the chemical nature of the resin, the surface finish of the resin, and the molecular structure of the LPS.

N-nitrosonornicotine (NNN), a significant nitrosamine component of tobacco binds to and is taken up by cells prior to its activation by intracellular enzymes. A variety of factors may affect the binding of substances to a cell including the cell type, medium composition and the cell membrane lipid composition. This study examines the binding and uptake of NNN to various cell types and relates these to the cells' lipid composition. Hamster buccal pouch keratinocytes were exposed to NNN and the amount of bound NNN determined. This was compared with cells pretreated with 12-0-tetradecanoylphorbol-13-acetate (TPA). While the keratinocytes bound significant amounts of NNN, this binding was not specific. Pretreatment of the cells with TPA significantly increased the amount of NNN bound. Cultures of human gingival fibroblasts and a human liver cell line also bound NNN, in quantities greater than that bound to the keratinocytes. The TPA caused changes in long chain unsaturated fatty acids although major changes in lipid metabolism were not detected. The fatty acid composition of the liver cells more closely resembled that of the TPA treated keratinocytes than the untreated ones. The data suggest that NNN binds to the cells non-specifically and the binding may be related to the amount of long-chain, unsaturated fatty acids present.