Specific differences in migratory function of myofibroblasts isolated from Crohn's disease fistulae and strictures.
BACKGROUND
Recently we found that migration of colonic lamina propria fibroblasts in Crohn's disease patients (CD-CLPF) from inflamed mucosa is significantly reduced as compared to control-CLPF. The behavior of CD-CLPFs isolated from fistulae and strictures was now investigated in detail.


METHODS
Initially migration assays for all CLPF cultures (CD-CLPF, fibrosis-CLPF, and fistula-CLPF) were performed in the modified 48-well Boyden chamber. Subsequently, for a migration assay more resembling the in vivo situation a 3D matrix model was developed. After seeding of cells into the 3D matrix the CLPF layer was wounded by an ERBIUM:YAG laser leading to circular cell rupture without effect on the extracellular matrix.


RESULTS
In the modified Boyden chamber migration of fistula-CLPF was significantly reduced compared to CD-CLPF. This was correlated with a decrease in FAK-protein expression, whereas in migrating fibrosis-CLPF an increase in FAK-protein expression, -autophosphorylation and migratory potential was found. This was confirmed in the 3D matrix wounding assay: Fistula-CLPF migrated less than CD-CLPF, whereas fibrosis-CLPF migrated significantly more in the 3D matrix wounding assay. Between 1 to 36 hours incubation time fibrosis-CLPF always displayed increased migration ability as compared to CD-CLPF. In contrast, fistula-CLPF migratory potential was always below that of CD-CLPF.


CONCLUSIONS
Myofibroblasts isolated from inflamed, fibrostenotic, or fistulized CD mucosa differ in their migratory potential both in the modified Boyden chamber as well as in a 3D matrix model. These different migratory behaviors could be an explanation for impaired or excess wound healing and subsequently for fistula and fibrosis formation.
Meier
Johannes K-H
JK
Department of Cranio-Maxillo-Facial Surgery, University of Regensburg, Germany.
Scharl
Michael
M
Miller
Sandra N
SN
Brenmoehl
Julia
J
Hausmann
Martin
M
Kellermeier
Silvia
S
Schölmerich
Jürgen
J
Rogler
Gerhard
G
D000328:Adult; D015153:Blotting, Western; D002465:Cell Movement; D002478:Cells, Cultured; D003251:Constriction, Pathologic / Q000209:etiology* / Q000378:metabolism / Q000473:pathology; D003424:Crohn Disease / Q000150:complications* / Q000473:pathology / Q000628:therapy; D005260:Female; D005355:Fibrosis / Q000209:etiology* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism / Q000473:pathology*; D008297:Male; D008875:Middle Aged; D009092:Mucous Membrane / Q000378:metabolism / Q000473:pathology*; D058628:Myofibroblasts / Q000378:metabolism / Q000473:pathology*; D011379:Prognosis; D014945:Wound Healing