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Matrix Biol. 2003 Nov;22(6):477-90.

ECM gene expression correlates with in vitro tissue growth and development in fibrin gel remodeled by neonatal smooth muscle cells.

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  • 1Departments of Biomedical Engineering, University of Minnesota, Minneapolis, MN 55455, USA.

Abstract

A tissue growth and development process occurred in neonatal SMC-fibrin gel constructs when cultured in DMEM supplemented with TGF-beta1 and insulin over a 5 week period. These constructs may thus serve as the basis for cardiovascular tissue replacements and future models of cardiovascular tissue growth, repair and regeneration. Following fibrin gel contraction during week 1, peak rates of SMC proliferation, collagen production and tropoelastin production occurred between weeks 1-4. Organized, cross-linked collagen and elastic fibers replaced the degrading fibrin over weeks 3-5 and were manifested as increased mechanical strength. The peak rate of SMC proliferation (weeks 1-2) preceded that for maximum collagen production (weeks 2-4), which was consistent with the 3 week time point of maximum expression of collagen type I and III from qRT-PCR. Insoluble elastin quantification revealed that the majority of elastic fibers were produced by week 4, which was also consistent with the qRT-PCR data showing a dramatic down-regulation of tropoelastin expression by week 4, indicating elastogenesis occurred during the early stages of tissue growth and development. There was a strong up-regulation of lysyl oxidase expression during weeks 1-3 with a peak in expression at week 3, correlating with the phases of collagen and tropoelastin production. An increase in MMP-2 expression over weeks 1-5 suggested an increase in ECM remodeling as the tissue developed. Mechanical strength doubled over weeks 4-5 when production of collagen and elastic fibers and expression of lysyl oxidase were subsiding. This may have been due in part to the more organized collagen fibrils evident from the histological sections in weeks 3-5.

PMID:
14667840
[PubMed - indexed for MEDLINE]
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