Keratinocyte-fibroblast paracrine interaction: the effects of substrate and culture condition.
Interactions between epidermal-dermal cells via soluble factors provide important signals in regulating the reepithelialization of wounded skin. For example, keratinocytes regulate the expression of keratinocyte growth factor (KGF) in fibroblasts through the release of interleukin-1beta (IL-1beta). In this study, a previously developed polyethyleneglycol-based interpenetrating network (IPN) system was utilized as a platform for the delivery of keratinocyte-active factors. The effect of substrate chemistry, culture condition, and the delivery of exogenous keratinocyte-active factors on the keratinocyte behavior and the keratinocyte-fibroblast paracrine relationship was delineated. Adherent keratinocyte density on TCPS and glutaraldehyde-fixed gelatin hydrogels but not on IPN was significantly increased with culture time in the presence of growth supplements independent of the released KGF from the gelatin hydrogel and IPN. In the presence of fibroblasts, adherent keratinocyte density on gelatin hydrogels was higher than that without fibroblasts. This phenomenon was not observed on IPN and polycarbonate membrane. In summary, the delivered exogenous huKGF (i.e., released from a biomaterial matrix) operates in tandem with fibroblasts in regulating keratinocyte activation (i.e., IL-lbeta release and adhesion) in a surface-dependent manner. Immunoassay analysis of cell culture keratinocyte-fibroblast paracrine relationship as characterized by IL-1beta and KGF could not be established in the presence of IPNs, 0.1% glutaraldehyde-fixed gelatin hydrogels, and polycarbonate membranes.
Witte
Richard P
RP
Department of Biomedical Engineering, College of Engineering, University of Wisconsin-Madison, WI, USA.
Kao
Weiyuan John
WJ
D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D018929:Cell Culture Techniques / Q000379:methods; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D020099:Coated Materials, Biocompatible / Q000008:administration & dosage / Q000737:chemistry; D018920:Coculture Techniques / Q000379:methods; D016503:Drug Delivery Systems / Q000379:methods; D051523:Fibroblast Growth Factor 7; D005346:Fibroblast Growth Factors / Q000008:administration & dosage*; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects*; D006801:Humans; D015603:Keratinocytes / Q000166:cytology* / Q000187:drug effects*; D008422:Materials Testing; D019899:Paracrine Communication / Q000187:drug effects / Q000502:physiology*; D017768:Skin, Artificial; D023822:Tissue Engineering / Q000379:methods*
D020099:Coated Materials, Biocompatible; C496453:FGF7 protein, human; D051523:Fibroblast Growth Factor 7; D005346:Fibroblast Growth Factors