Decreased levels of alpha 1(I) procollagen mRNA in dermal fibroblasts grown on fibrin gels and in response to fibrinopeptide B.
We have investigated human neonatal fibroblast synthetic activity in response to fibrin substrates and components of fibrin formation and degradation. Greater than threefold downregulation of procollagen mRNA levels was seen 24 hours after fibroblasts were grown on fibrin gels as compared to tissue culture plastic. This downregulation occurred in both reptilase-generated fibrin (retention of fibrinopeptide B) and thrombin-generated fibrin (loss of both fibrinopeptide A and B). However, fibroblasts grown on fibrin retained their capacity to respond to the stimulatory action of transforming growth factor (TGF)-beta 1. Fibroblasts seeded on reptilase-generated fibrin displayed an abnormal morphology manifested by dendritic appearance and cell rounding, while fibroblast attachment was enhanced by 30% on thrombin-generated fibrin substrate (P < 0.02). Fibrinopeptides A and B, which are generated during fibrin formation, increased and decreased procollagen mRNA levels, respectively. Tissue plasminogen activator (t-PA) increased procollagen mRNA and TGF-beta 1 levels as early as 6 hours after cells were grown on tissue culture plastic, but this stimulation did not occur in cells cultured on a fibrin substrate. We conclude that alpha 1(I) procollagen mRNA levels in cultures of human dermal fibroblasts are consistently down-regulated by a fibrin substrate and are directly and profoundly influenced by complex interactions between components involved in the formation and removal of fibrin.
Pardes
J B
JB
University of Miami School of Medicine, Department of Dermatology and Cutaneous Surgery, Florida 33136.
Takagi
H
H
Martin
T A
TA
Ochoa
M S
MS
Falanga
V
V
D015152:Blotting, Northern; D002478:Cells, Cultured; D015536:Down-Regulation; D005337:Fibrin / Q000032:analysis / Q000378:metabolism / Q000494:pharmacology*; D005344:Fibrinopeptide A / Q000032:analysis / Q000378:metabolism / Q000494:pharmacology; D005345:Fibrinopeptide B / Q000032:analysis / Q000378:metabolism / Q000494:pharmacology*; D005347:Fibroblasts / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism; D005782:Gels; D006801:Humans; D007231:Infant, Newborn; D008297:Male; D011347:Procollagen / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D010959:Tissue Plasminogen Activator / Q000032:analysis / Q000378:metabolism / Q000494:pharmacology; D016212:Transforming Growth Factor beta / Q000032:analysis / Q000378:metabolism / Q000494:pharmacology
D005782:Gels; D011347:Procollagen; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; D005344:Fibrinopeptide A; D005345:Fibrinopeptide B; D005337:Fibrin; D010959:Tissue Plasminogen Activator