In vitro pharmacological inhibition of myofibroblast differentiation and force generation in a collagen-GAG matrix

Induced regeneration studies from three organs in the adult mammal (skin, peripheral nerves, and the conjunctiva) suggest an antagonistic relationship between myofibroblast-mediated contraction of wounds and induced regeneration. In each instance of induced regeneration, contraction blocking was accomplished using regeneration templates, or scaffolds of highly specific structural and chemical properties that mainly control the environment or the density of contractile cells (myofibroblasts). In addition to scaffolds, diffusible factors could be used to inhibit specific components of the intracellular pathway in an effort to further evaluate the relationship between contraction and induced regeneration. The crucial role that Rho- associated coiled-coil forming protein serine/threonine kinase (ROCK) seems to play in cell-mediated contraction and cytoskeletal remodeling behavior of fibroblasts make it an attractive target for pharmacological inhibition. A preliminary in vitro study was conducted to evaluate the effect of Y-27632, a specific pharmacological inhibitor of ROCK, on the contraction of highly porous, three-dimensional type I collagen- glycosaminoglycan (CG) matrices over time by attached NR6WT fibroblasts treated with TGF-[beta]1, a known upregulator of both fibroblast contraction and expression of the contractile filament [alpha]-smooth muscle actin ([alpha]-SMA), a hallmark of the myofibroblast phenotype. NR6WT fibroblasts were seeded into free-floating cylindrical CG matrices (8 mm in diameter, n=6) and treated with serum-containing media supplemented with TGF-[beta]1 (3 ng/ml) or both TGF-[beta]1 (3 ng/ml) and Y--27632 (10 [mu]M) for 12 days.

Department dc:contributor.department

Massachusetts Institute of Technology. Dept. of Mechanical Engineering.

Grantor dc:publisher

Massachusetts Institute of Technology

Year dc:date.issued

2005

Author dc:creator

• Soller, Eric C

Advisor dc:contributor.advisor

• Ioannis V. Yannas.

• Mechanical Engineering.

Statement dc:rights

• M.I.T. theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission. See provided URL for inquiries about permission.

Licence dc:rights.uri

• http://dspace.mit.edu/handle/1721.1/7582

Language dc:language.iso

eng

Handle dc:identifier.uri

http://hdl.handle.net/1721.1/32343

OAI identifier oai:identifier

oai:dspace.mit.edu:1721.1/32343