Adherence, proliferation and collagen turnover by human fibroblasts seeded into different types of collagen sponges.
We describe an in vitro model that we have used to evaluate dermal substitutes and to obtain data on cell proliferation, the rate of degradation of the dermal equivalent, contractibility and de novo synthesis of collagen. We tested three classes of collagenous materials: (1) reconstituted non-crosslinked collagen, (2) reconstituted collagen that was chemically crosslinked with either glutaraldehyde, aluminium alginate or acetate, and (3) native collagen fibres, with or without other extracellular matrix molecules (elastin hydrolysate, hyaluronic acid or fibronectin). The non-crosslinked reconstituted collagen was degraded rapidly by human fibroblasts. The chemically crosslinked materials proved to be cytotoxic. Native collagen fibres were stable. In the absence of ascorbic acid, the addition of elastin hydrolysate to this type of matrix reduced the rate of collagen degradation. Both elastin hydrolysate and fibronectin partially prevented fibroblast-mediated contraction. Hyaluronic acid was only slightly effective in reducing the collagen degradation rate and more fibroblast-mediated contraction of the material was found than for the native collagen fibres with elastin hydrolysate and fibronectin. In the presence of ascorbate, collagen synthesis was enhanced in the native collagen matrix without additions and in the material containing elastin hydrolysate, but not in the material with hyaluronic acid. These results are indicative of the suitability of tissue substitutes for in vivo application.
Middelkoop
E
E
Department of Cell Biology and Histology, Academic Medical Center, Amsterdam, The Netherlands.
de Vries
H J
HJ
Ruuls
L
L
Everts
V
V
Wildevuur
C H
CH
Westerhof
W
W
D001205:Ascorbic Acid / Q000494:pharmacology; D002448:Cell Adhesion; D002455:Cell Division; D002478:Cells, Cultured; D003094:Collagen / Q000187:drug effects / Q000378:metabolism; D003432:Cross-Linking Reagents / Q000494:pharmacology; D046508:Culture Techniques / Q000295:instrumentation*; D004549:Elastin / Q000494:pharmacology; D005109:Extracellular Matrix / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D005353:Fibronectins / Q000494:pharmacology; D006801:Humans; D006820:Hyaluronic Acid / Q000494:pharmacology; D008855:Microscopy, Electron, Scanning; D017768:Skin, Artificial; D015503:Surgical Sponges
D003432:Cross-Linking Reagents; D005353:Fibronectins; D006820:Hyaluronic Acid; D003094:Collagen; D004549:Elastin; D001205:Ascorbic Acid