Specific differences in migratory function of myofibroblasts isolated from Crohn's disease fistulae and strictures.

Abstract

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.

• Authors: Johannes K-H Meier, Michael Scharl, Sandra N Miller, Julia Brenmoehl, Martin Hausmann, Silvia Kellermeier, Jürgen Schölmerich, Gerhard Rogler
• Journal: Inflammatory bowel diseases (Inflamm Bowel Dis) Vol. 17 Issue 1 Pg. 202-12 (Jan 2011) ISSN: 1536-4844 [Electronic] England
• PMID: 20848526 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2010 Crohn's & Colitis Foundation of America, Inc.

Topics

• Adult
• Blotting, Western
• Cell Movement
• Cells, Cultured
• Constriction, Pathologic (etiology, metabolism, pathology)
• Crohn Disease (complications, pathology, therapy)
• Female
• Fibrosis (etiology, metabolism, pathology)
• Humans
• Intestinal Mucosa (metabolism, pathology)
• Male
• Middle Aged
• Mucous Membrane (metabolism, pathology)
• Myofibroblasts (metabolism, pathology)
• Prognosis
• Wound Healing