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Effect of Initial Collagen Concentration on Fibroblast Mediated Contraction of Collagen Gels* FREE TO VIEW

Yun Kui Zhu, MD; T. Umino, MD; X.D. Liu, MD; H.J. Wang, MD; Debra J. Romberger, MD, FCCP; John R. Spurzem, MD, FCCP; Stephen I. Rennard, MD, FCCP
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*From the Pulmonary and Critical Care Medicine Section, University of Nebraska Medical Center, Omaha, NE.

Correspondence to: Yun Kui Zhu, MD, Pulmonary and Critical Care Medicine Section, Department of Internal Medicine, University of Nebraska Medical Center, 600 South 42nd Street, Omaha, NE 68198-5300.

Chest. 2000;117(5_suppl_1):234S-235S. doi:10.1378/chest.117.5_suppl_1.234S
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Remodeling of extracellular matrix involves a number of steps, including the recruitment, accumulation, and eventual apoptosis of parenchymal cells and the production, organization, and rearrangement of extracellular matrix produced by these cells. The culture of fibroblasts in three-dimensional gels made of type I collagen has been used as a model of tissue re-arrangement. Specifically, contraction of collagen gels is believed to resemble the contraction that characterizes both wound repair and fibrosis. The current study was designed to determine the effect of initial collagen concentration on the final size of contracting collagen gels and on the final number of fibroblasts present within the gels. Native type I collagen was extracted from rat tail tendons and was used to prepare collagen gels with varying collagen concentrations. Fibroblasts were cast into these gels. Gels prepared with an initial concentration of 0.75 mg/mL contracted more rapidly and to a smaller final size than did gels prepared from 2 mg initial collagen concentration (final size 7.1 vs 36.43% of initial size; p < 0.01). There was no significant degradation of the collagen gel under either condition. Hence, the dramatically increased contraction in the lower density gels resulted in a higher final density (p < 0.01). Cell density was estimated from DNA content. In low-density gels, DNA was significantly less than that in higher density gels (0.73 vs 1.88 μg; p < 0.01). This was accompanied by an increased percentage of apoptotic cells in the low-density collagen gels at day 14 (43.3 vs 34.1; p < 0.05). If the gels were maintained in the attached state, which largely prevents contraction, apoptosis was significantly reduced, suggesting that contraction rather than matrix composition was a requirement for the increased apoptosis. Taken together, these findings indicate that the initial matrix composition can lead to differing outcomes during fibroblast mediated contraction of collagen gels. Such effects may contribute to the altered tissue structures that characterize the lung in COPD.




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