COL1A1 oligodeoxynucleotides decoy: biochemical and morphologic effects in an acute wound repair model.
Type I collagen is an integral component of granulation tissue and scar, that is highly dependent on TGFβ1, a member of a pro-fibrotic family of cytokines, for its promotion and deposition. Blocking COL1A1 gene transcription obstructs type I collagen synthesis, hindering the progress of granulation tissue deposition and fibrosis. Local injections of a double stranded oligodeoxynucleotide (dsODN) decoy, containing the TGFβ1 regulatory element that is located in the distal promoter of the COL1A1 gene, were investigated in a rat polyvinyl alcohol (PVA) sponge granulation tissue implant model. The effects on the granulation tissue deposition by dsODN decoy therapy were evaluated by the synthesis of types I and III collagens as well as ED-A (cellular) fibronectin. Fluorescently labeled dsODN was used to identify the distribution of the decoy molecules in the sponge implant relative to the observed histological effects. Morphological alterations in cells and changes in the organization of connective tissue were documented and evaluated. Collagen levels were reduced by half in implants treated with 10 nM dsODN decoy compared to scrambled dsODN-treated implants. Histologically, dsODN decoy treated implants had an increased cellular density without a corresponding increase in deposited connective tissue. Polarized light birefringence pattern of Sirius red-stained sections showed less collagen fibers accumulating between fibroblasts. The highest concentration of fluorescently labeled dsODN was identified within the interior margin of sponge implants, correlating to increased cellular density and an altered birefringence patterns. In conclusion, 10 nM dsODN decoy therapy reduced collagen deposition and altered the organization of granulation tissue, supporting its potential as a localized anti-fibrotic therapy for limiting fibrotic conditions.
Rosensteel
Shawn M
SM
Department of Comparative Medicine, The Pennsylvania State University, College of Medicine, Hershey, PA, USA. shr26@pitt.edu
Wilson
Ronald P
RP
White
Sheryl L
SL
Ehrlich
H Paul
HP
D000818:Animals; D002921:Cicatrix / Q000378:metabolism / Q000517:prevention & control; D024042:Collagen Type I / Q000037:antagonists & inhibitors*; D000090602:Collagen Type I, alpha 1 Chain; D003238:Connective Tissue / Q000378:metabolism*; D005355:Fibrosis / Q000378:metabolism / Q000517:prevention & control; D006097:Granulation Tissue / Q000378:metabolism; D015151:Immunoblotting; D008856:Microscopy, Fluorescence; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D050436:Regulatory Elements, Transcriptional / Q000235:genetics; D053773:Transforming Growth Factor beta1 / Q000235:genetics; D014945:Wound Healing / Q000502:physiology*
D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; D020319:Oligodeoxyribonucleotides, Antisense; C508962:Tgfb1 protein, rat; D053773:Transforming Growth Factor beta1