Cocktail Treatment of Calcium Channel Blocker, Steroid and Interferon to Affect Transforming Growth Factor Beta (TGF-β) and Basic Fibroblast Growth Factor (bFGF) to Control Hypertrophic Scar Formation in Human Scar Carrying Animal Model

The formation of hypertrophic scar(HSC) is an abnormal wound-healing response after skin damage including thermal injuries . Because its pathogenesis is not clearly understood, it still cannot be controlled with satisfying results.The previous study revealed that extracellular matrix molecules, cytokines, and proteinases may alter cell proliferation and migration and thus lead to the formation of hypertrophic scar. The mechanisms may relate to the inhibition of dermal fibrosis, the modulation of Basic fibroblast growth factor(bFGF)、Transforming growth factor beta (TGF-β) to regulate the fibroblast proliferation or differentiation. Besides, induce apoptosis can also inhibit the scar formation. However, on the basis of the view of modern biology, any change in response to an injury must be accompanied by a change in gene expression. In the previous study, these altered genes were related to proto-oncogenes, apoptosis, immune regulatory genes, cytoskeletal elements, metabolism, and so forth. So we evaluate the gene expressions in the formation of the scars may be valuable for finding scar-related genes, for understanding the pathologic alteration, specific marker and the therapeutic strategy. In our previous study (NSC89-2314-B-182A-151, NSC94-2314-B-182A-179), the animal model with human scar implanted into nude mice (BALB/c) has been established successfully and the effects of combination therapy with verapamil and kenacort have been studied and compared both in vitro and in vivo after 4 weeks treatment. In the aspects of scar size and weight, decorin stain, fibroblast culture to detect the fibroblast activity, the therapeutic groups have much better response. The paper has been publicated by journal Dermatologic Surgery. {appendix 1(Ref.1 )} There are some mechanism involving HSC formation. One is high fibroproliferative activity resulting in excessive collagen accumulation. Another is disorganized collagen fibers due to reduction in some proteoglycan such as decorin. The expression of decorin and collagenase genes of HSC fibroblasts can be blocked by some cytokines such as TGF-β(Scott, 1998). Our previous study has indirectly proved that corticosteroids, calcium channel blockers(CCB) and Interferon(IFN-α) can restore the synthesis of decorin and collagenase which has been retarded by these cytokines. In the fallowing study we get the results that the combination therapy can decrease fibroblast proliferation, reduce contraction power (Fibroblast-Populated Collagen Lattice) and collagen amount. Then the results were approved by another study(NSC98-2314-B- 182 -017) to show that the combination therapy can increase the gene expression of collagenase(MMP13) and Decorin. All were prevented at international conference (Ref.2).However the studies mentioned above seems concentrated on the cell and enzyme level. Because during the maturation phase the scar formation still is affected by some cytokines such as bFGF、 TGF-β. This study is designed to understand the mechanism, function of combination therapy to affect the cytokines and then to control scars. In the first year, we will divide the human-scar carrying nude mice into three group. One with no injection, one with N.S. injection and the 3rd with cocktail{steroid(40mg/c.c)0.04c.c + CCB(2.5mg/c.c)0.06c.c + IFN-α(15 million I.U/ml)0.02c.c} injection to scars. Before injection and one month after injection, the fibroblast culture from scars will be used for ELISA,Western Blot Analysis to detect the amount of TGF-β1、TGF-β3 and bFGF. Hope we can approve that the effects of cocktail regimen is also going through cytokine mechanism. In this study we also can compare the difference of injection and no injection of cocktail regimen. In the second year, fibrocyte study will be the key point. We also use MMP13 detection, Fibroblast- Populated Collagen Lattice (FPCL) study to approve the possible mechanism that is cocktail regimen may increase TGF-β3, inhibit fibrocyte and increase MMP13 expression and FPCL effect.

Project ID:PC10207-0416
External Project ID:NSC102-2314-B182A-117