TGF-β Antisense Therapy Increases Angiogenic Potential in Human Keratinocytes In Vitro∗

BackgroundTransforming growth factor-beta (TGF-β) has been identified as an important component of wound healing. Recent developments in molecular therapy offer exciting prospects for the modulation of wound healing, specifically those targeting TGF-β. The purpose of this study was to analyze the effect of TGF-β targeting on the expression of angiogenic vascular endothelial growth factor (VEGF), a key regulator of angiogenesis, and in vitro angiogenic activity.MethodsExpression of angiogenic VEGF in tissue samples from chronic dermal wounds was investigated by immunohistochemistry. The effect of TGF-β targeting using antisense oligonucleotides on the expression of VEGF was analyzed by ELISA and RT-PCR in cultured human keratinocytes. Human endothelial cells (EC) were grown in conditioned medium produced from the treated keratinocytes. EC migration was measured using a modified Boyden chamber, EC tube formation was analyzed under the light microscope.ResultsImmunohistochemical investigation demonstrated a decreased expression of VEGF protein in tissue samples from chronic dermal wounds compared to normal human skin. Antisense TGF-β oligonucleotide treatment upregulated VEGF secretion in vitro. Addition of conditioned medium from TGF-β antisense-treated keratinocytes resulted in an increase of endothelial cell migration and tube formation.ConclusionsOur results demonstrate that TGF-β antisense oligonucleotide technology may be a potential therapeutic option for stimulation of angiogenesis in chronic wounds.