ECM gene expression correlates with in vitro tissue growth and development in fibrin gel remodeled by neonatal smooth muscle cells
ECM gene expression correlates with in vitro tissue growth and development in fibrin gel remodeled by neonatal smooth muscle cells
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DOI：
10.1016/s0945-053x(03)00078-7
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发表时间：
2003-11-01
期刊：
影响因子：
6.9
通讯作者：
Tranquillo, R
中科院分区：
文献类型：
--
作者：
Ross, JJ;Tranquillo, R
新生儿SMC-纤维蛋白凝胶构建体在补充有TGF-β 1和胰岛素的DMEM中培养超过5周时发生组织生长和发育过程。因此，这些构建体可以作为心血管组织替代物和心血管组织生长、修复和再生的未来模型的基础。在第1周纤维蛋白凝胶收缩后，SMC增殖、胶原蛋白产生和原弹性蛋白产生的峰值发生在第1-4周之间。在3-5周内，有组织的交联胶原蛋白和弹性纤维取代了降解的纤维蛋白，并表现为机械强度增加。SMC增殖的峰值速率（第1-2周）先于最大胶原蛋白产生的峰值速率（第2-4周），这与qRT-PCR中I型和III型胶原蛋白最大表达的3周时间点一致。不溶性弹性蛋白定量显示，大多数弹性纤维在第4周产生，这也与qRT-PCR数据一致，其显示到第4周弹性蛋白原表达显著下调，表明弹性蛋白发生在组织生长和发育的早期阶段。赖氨酰氧化酶表达在第1-3周期间有强烈的上调，在第3周表达达到峰值，与胶原蛋白和原弹性蛋白产生的阶段相关。MMP-2表达在1-5周内的增加表明随着组织发育ECM重塑增加。当胶原蛋白和弹性纤维的产生以及赖氨酰氧化酶的表达消退时，机械强度在4-5周内翻了一番。这可能部分是由于在第3-5周的组织学切片中明显存在更有组织的胶原纤维。(C)2003年Elsevier B. V./国际矩阵生物学学会。All rights reserved.
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. (C) 2003 Elsevier B.V./International Society of Matrix Biology. All rights reserved.