Effect of transforming growth factor-β1 antisense oligonucleotides on matrix metalloproteinases and their inhibitors in keloid fibroblasts

Abstract

Objective To identify changes in the expression of matrixmetalloproteinases (MMPs) and their specific inhibitors tissue inhibitors of metalloproteinases (TIMPs) after targeting of transforming growth factor-β1 (TGF-β1) with antisense oligonucleotides. Study Design Cross-sectional study. Setting The study was performed on tissue samples from nine patients with keloid scars after otoplasty presenting to the Otolaryngology–Head and Neck Surgery Department of the University Hospital in Mannheim, Germany. Subjects and Methods Keloid fibroblasts and normal fibroblasts were harvested from auricular keloid scars and healthy skin regions of the same patients during resection procedure of the keloid. Cells were placed in monolayer cultures. Expression of MMPs and TIMPs were analyzed by immunohistochemistry. The effect of TGF-β1 targeting using antisense oligonucleotides on the expression of both protein groups in keloid-derived fibroblasts was analyzed by enzyme-linked immunosorbent assay and reverse-transcription polymerase chain reaction. Results Immunohistochemical investigation demonstrated increased expression of MMP-2, -3, -9, and -13 and TIMP-1 and -2. TGF-β1 antisense therapy significantly down-regulated MMP secretion in vitro. Conclusion Usage of TGF-β1 antisense oligodeoxynucleotides (ODNs) may show a potential chemopreventive or therapeutic option for keloids by blocking the effect of TGF-β1. Furthermore, antisense ODNs can be used as an investigative approach toward a better understanding of molecular mechanisms in keloid pathophysiology.