Fibronectin and its alpha 5 beta 1-integrin receptor are involved in the wound-repair process of airway epithelium.
The cell migration that occurs during wound repair is dependent on modifications of the cell-matrix interaction in which extracellular matrix proteins and their receptors, the integrins, are involved. To study the interactions between airway epithelial cells and the extracellular matrix during the process of wound repair, we developed an in vitro wound model of human epithelial cells. Surface epithelial cells were dissociated from human nasal polyps and cultured on a type I collagen matrix. At confluency, a wound was made by the addition of 2 microliters of NaOH (1 N) to the cell culture. After the cell culture was washed, the wound area was recorded every 12 h for 96 h by a videomicroscopic technique. We calculated the wound-repair index that represents the decrease in the wound area per hour. Using immunofluorescence techniques, we first examined the localization, during wound repair, of fibronectin and of the beta 1-, alpha v-, alpha 2-, alpha 3-, and alpha 5-integrin subunits. Secondly, we carried out a series of wound-repair blocking experiments with the use of anti-integrin or anti-fibronectin antibodies diluted in the culture medium. We observed that fibronectin and the alpha 5- integrin subunit were exclusively expressed by the migratory cells in the wounded area. No difference in the localization of the alpha v-, alpha 2-, and alpha 3-integrin subunits was observed between the nonrepairing and repairing cells. The blocking experiments showed a significant decrease in the wound-repair index in the presence of either the anti-beta 1, -alpha 3, alpha 5, or the anti-fibronectin antibodies. Furthermore, the addition of fibronectin to the culture medium induced a significant increase in the wound repair index. These results suggest that fibronectin and the corresponding alpha 5 beta 1-integrin play an important role in the process of airway epithelium wound repair.
Hérard
A L
AL
Institut National de la Santé et de la Recherche Médicale U314, Reims France.
Pierrot
D
D
Hinnrasky
J
J
Kaplan
H
H
Sheppard
D
D
Puchelle
E
E
Zahm
J M
JM
D000906:Antibodies / Q000494:pharmacology; D002465:Cell Movement; D002478:Cells, Cultured; D004847:Epithelial Cells; D004848:Epithelium / Q000502:physiology; D005353:Fibronectins / Q000032:analysis / Q000276:immunology / Q000502:physiology*; D006801:Humans; D007150:Immunohistochemistry; D009298:Nasal Polyps / Q000473:pathology / Q000503:physiopathology* / Q000601:surgery; D017469:Receptors, Fibronectin / Q000032:analysis / Q000276:immunology / Q000502:physiology*; D014945:Wound Healing
D000906:Antibodies; D005353:Fibronectins; D017469:Receptors, Fibronectin