Human dermal and gingival fibroblasts in a three-dimensional culture: a comparative study on matrix remodeling.
Free-floating collagen lattice is considered a useful tool for assessing wound healing in vitro. This work compared extracellular matrix remodeling in collagen lattices populated by gingival or dermal fibroblasts. For 21 days we followed gel contraction and changes in cell number of collagen lattices seeded with l.5 x 10(5) fibroblasts of each tissue. We also used indirect immunodetection to study extracellular matrix components, metalloproteinases (MMPs), and their tissues inhibitors (TIMPs). In addition, the presence of MMPs and TIMPs in the culture media was analyzed by zymography and western blotting. No significant difference was found concerning gel contraction and changes in cell number. We observed the early expression of fibrillin I and collagen type III, apparently codistributed and at the end of the gel contraction their disappearance. Concomitantly we demonstrated the expression of MMPs and TIMPs, initially localized in cellular cytoplasm, then spreading in the extracellular compartment, and even found in the culture medium. This remodeling was more rapid and intense with gingival fibroblasts than dermal fibroblasts. In conclusion, gingival fibroblasts seem more efficient at remodeling the connective tissue than dermal fibroblasts and could lead to the better wound healing observed in vivo.
Chaussain Miller
C
C
Laboratoire de Physiopathologie des Tissus non Mineralises, Faculté de Chirurgie Dentaire Paris V, Montrouge, France. miller@odontologie.univ-paris5.fr
Septier
D
D
Bonnefoix
M
M
Lecolle
S
S
Lebreton-Decoster
C
C
Coulomb
B
B
Pellat
B
B
Godeau
G
G
D000293:Adolescent; D000328:Adult; D015153:Blotting, Western; D002452:Cell Count; D002478:Cells, Cultured; D002648:Child; D018920:Coculture Techniques; D003094:Collagen / Q000378:metabolism; D024042:Collagen Type I / Q000032:analysis; D024061:Collagen Type III / Q000032:analysis; D003239:Connective Tissue Cells / Q000378:metabolism; D003593:Cytoplasm / Q000201:enzymology; D020405:Dermis / Q000166:cytology*; D004549:Elastin / Q000032:analysis; D005109:Extracellular Matrix / Q000378:metabolism*; D016326:Extracellular Matrix Proteins / Q000032:analysis; D000071837:Fibrillins; D005347:Fibroblasts / Q000378:metabolism*; D016650:Fluorescein-5-isothiocyanate; D019084:Fluorescent Antibody Technique, Indirect; D005456:Fluorescent Dyes; D005881:Gingiva / Q000166:cytology*; D006801:Humans; D020782:Matrix Metalloproteinases / Q000032:analysis; D008840:Microfilament Proteins / Q000032:analysis; D008854:Microscopy, Electron; D019714:Tissue Inhibitor of Metalloproteinases / Q000032:analysis; D014945:Wound Healing
D024042:Collagen Type I; D024061:Collagen Type III; D016326:Extracellular Matrix Proteins; D000071837:Fibrillins; D005456:Fluorescent Dyes; D008840:Microfilament Proteins; D019714:Tissue Inhibitor of Metalloproteinases; D003094:Collagen; D004549:Elastin; D020782:Matrix Metalloproteinases; D016650:Fluorescein-5-isothiocyanate