Human renal fibroblast contraction of collagen I lattices is an integrin-mediated process.
BACKGROUND
Expression of the beta1 family of integrins allows dermal fibroblasts in wounds to contribute to the healing process through migration, adhesion, synthesis, and rearrangement of extracellular matrix. To date the ability of human renal fibroblasts to reorganize collagens and the role of cell surface receptors in this process remain unknown.


METHODS
Renal fibroblasts were grown from the cortical tissue of surgically removed human kidneys. The ability of human renal fibroblasts to reorganize interstitial collagen I was examined in vitro using solidified collagen I lattices. Integrin function was blocked by incubating fibroblasts with isotype-specific antibodies prior to addition to collagen I lattices.


RESULTS
Human renal fibroblasts embedded in collagen I lattices progressively decreased lattice diameter to 60.6+/-11.4% of initial diameter at 48 h post-release (P:<0.01). Fibroblasts incubated in the presence of antibody to beta1 integrin failed to contract collagen I lattices, whilst fibroblasts incubated with non-specific antibody reduced lattice diameter to 60.1+/-12.4% of initial diameter at 48 h post-release (P:<0.01). Further characterization of integrin alpha subunits showed that blocking alpha2beta1 integrin prevented lattice contraction (P:<0.05, alpha2beta1 integrin antibody vs non-specific antibody), whilst blocking of alpha5beta1, alpha3beta1 and alpha1beta1 integrins did not influence this process.


CONCLUSIONS
We postulate that collagen I fibril rearrangement by human renal fibroblasts in vitro appears to be an integrin-mediated process involving the alpha2beta1 integrin.
Kelynack
K J
KJ
Department of Nephrology, Royal Melbourne Hospital, Parkville and. Microvascular Biology and Wound Healing Group, RMIT University, Bundoora, Victoria, Australia.
Hewitson
T D
TD
Nicholls
K M
KM
Darby
I A
IA
Becker
G J
GJ
D000328:Adult; D000368:Aged; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D003094:Collagen; D005260:Female; D005347:Fibroblasts / Q000166:cytology* / Q000502:physiology; D006801:Humans; D039222:Integrin alpha1beta1; D039201:Integrin alpha3beta1; D019012:Integrin beta1 / Q000502:physiology; D016023:Integrins / Q000032:analysis / Q000502:physiology*; D007672:Kidney Cortex / Q000166:cytology* / Q000502:physiology; D008297:Male; D008875:Middle Aged; D038981:Receptors, Collagen; D017469:Receptors, Fibronectin / Q000502:physiology
D039222:Integrin alpha1beta1; D039201:Integrin alpha3beta1; D019012:Integrin beta1; D016023:Integrins; D038981:Receptors, Collagen; D017469:Receptors, Fibronectin; D003094:Collagen