Effect of transforming growth factor-beta1 antisense oligonucleotides on matrix metalloproteinases and their inhibitors in keloid fibroblasts.
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-beta1 (TGF-beta1) with antisense oligonucleotides.


METHODS
Cross-sectional study.


METHODS
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.


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-beta1 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-beta1 antisense therapy significantly down-regulated MMP secretion in vitro.


CONCLUSIONS
Usage of TGF-beta1 antisense oligodeoxynucleotides (ODNs) may show a potential chemopreventive or therapeutic option for keloids by blocking the effect of TGF-beta1. Furthermore, antisense ODNs can be used as an investigative approach toward a better understanding of molecular mechanisms in keloid pathophysiology.
Bran
Gregor M
GM
Department of Otolaryngology, Head and Neck Surgery, University Hospital of Mannheim, University of Heidelberg, Mannheim, Germany. gregor.bran@umm.de
Goessler
Ulrich R
UR
Baftiri
Ariton
A
Hormann
Karl
K
Riedel
Frank
F
Sadick
Haneen
H
D016022:Case-Control Studies; D018929:Cell Culture Techniques; D003430:Cross-Sectional Studies; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007627:Keloid / Q000209:etiology / Q000378:metabolism* / Q000473:pathology; D020782:Matrix Metalloproteinases / Q000235:genetics / Q000378:metabolism*; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology*; D013506:Otologic Surgical Procedures / Q000009:adverse effects; D012333:RNA, Messenger / Q000378:metabolism; D019714:Tissue Inhibitor of Metalloproteinases / Q000235:genetics / Q000378:metabolism*; D053773:Transforming Growth Factor beta1 / Q000235:genetics / Q000378:metabolism*
D020319:Oligodeoxyribonucleotides, Antisense; D012333:RNA, Messenger; D019714:Tissue Inhibitor of Metalloproteinases; D053773:Transforming Growth Factor beta1; D020782:Matrix Metalloproteinases