The motogenic effects of cyclic mechanical strain on intestinal epithelial monolayer wound closure are matrix dependent.
OBJECTIVE
Complex deformation during normal digestion due to peristalsis or villous motility may be trophic for the intestinal mucosa. Because tissue fibronectin is increased in inflammatory states that may accompany mucosal injury, we evaluated the effects of cyclic mechanical strain and fibronectin on intestinal epithelial monolayer wound closure in Caco-2 and IEC-6 intestinal epithelial cells.


METHODS
Wounds created in intestinal epithelial monolayers were subjected to cyclic deformation. Wound closure was assessed by morphometry using microscopic imaging. Cell signals were assessed by Western blot and confocal microscopy.


RESULTS
Mechanical strain stimulated wound closure on fibronectin but inhibited closure on collagen in Caco-2 and IEC-6 cells. The effect was independent of proliferation or cell spreading. Myosin light chain (MLC) and extracellular signal-regulated kinase (ERK) were phosphorylated in response to strain in confluent monolayers on both collagen and fibronectin. Blocking MLC or ERK phosphorylation inhibited the motogenic effect of strain on fibronectin. Although phosphorylated MLC was redistributed to the leading edge of migrating cells following 6 hours of strain on collagen and fibronectin, phosphorylated ERK was redistributed to the lamellipodial edge only on fibronectin.


CONCLUSIONS
Strain promotes intestinal epithelial wound closure by a pathway requiring ERK and MLC kinase. Fibronectin-dependent ERK redistribution in response to strain in confluent migrating cells may explain the matrix dependence of the motogenic effect. Repetitive deformation stimulates intestinal epithelial proliferation on a collagen substrate, but not fibronectin. Deformation may exert matrix-dependent effects on intestinal epithelial cells, promoting epithelial restitution in fibronectin-rich tissue and proliferation in fibronectin-poor mucosa.
Zhang
Jianhu
J
Department of Surgery, John D. Dingell VA Medical Center, 4646 John R. Street, Detroit, MI 48301, USA.
Owen
Cheri R
CR
Sanders
Matthew A
MA
Turner
Jerrold R
JR
Basson
Marc D
MD
D018938:Caco-2 Cells; D003094:Collagen; D005109:Extracellular Matrix / Q000201:enzymology*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005353:Fibronectins; D006801:Humans; D039564:Integrin alphaV / Q000378:metabolism; D007413:Intestinal Mucosa / Q000166:cytology / Q000502:physiology*; D018994:Myosin Light Chains / Q000378:metabolism; D010528:Peristalsis / Q000502:physiology*; D010766:Phosphorylation; D015398:Signal Transduction / Q000502:physiology*; D013314:Stress, Mechanical
D005353:Fibronectins; D039564:Integrin alphaV; D018994:Myosin Light Chains; D003094:Collagen; D048049:Extracellular Signal-Regulated MAP Kinases