2011 Volume 121 Issue 8 Pages 1607-1620
We examined the causes of the different effects of bFGF solution (dissolved in ultrapure water) and Fiblast® Spray solution (dissolved in attached lysate) at high concentrations (100–1,000 ng/ml) on the cell growth of normal human keratinocytes and fibroblasts cultured on type I collagen. We compared the effects of three kinds of bFGF solutions—a bFGF solution dissolved in ultrapure water at 0–1,000 ng/ml concentrations of bFGF and two solutions of Fiblast® Spray, one dissolved in attached lysate and the other in ultrapure water—on the cell growth of normal human keratinocytes and fibroblasts. We utilized a five-day cell growth assay to evaluate toxicity. We also tested the effects of the attached lysate only and benzalkonium chloride only by the same assay. The results showed that all three kinds of bFGF solutions accelerated the cell growth of both cell types in a concentration-dependent manner. For both cell types, the effects of bFGF solution and the Fiblast® Spray solution dissolved in ultrapure water were the same. However, the effect of Fiblast® Spray solution dissolved in attached lysate was somewhat different. When compared with the other two kinds of bFGF solution, there was an inhibition of cell growth, and there were also changes in cell morphology in both cell types at high concentrations (1,000 ng/ml with both cell types and also at 100 ng/ml with the keratinocytes) of bFGF. The same findings were observed with the addition of the medium that contained the same amount of attached lysate only, in which high concentrations of Fiblast® Spray solution were prepared, and also with the medium that contained benzalkonium chloride only. From these data, we concluded that Fiblast® Spray solution dissolved in attached lysate inhibited cell growth of both cell types at high concentrations (100–1,000 ng/ml), unlike the other two kinds of bFGF solutions dissolved in ultrapure water. The cause was the cytotoxicity of benzalkonium chloride in the attached lysate. This study demonstrated that benzalkonium chloride in the attached lysate should be avoided to get maximal acceleration of cell growth of the both cell types by Fiblast® Spray.
