Mechanoregulation of valvular interstitial cell phenotype in the third dimension.
Mechanoregulation of valvular interstitial cell phenotype in the third dimension.
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DOI：
10.1016/j.biomaterials.2013.10.047
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发表时间：
2014-01
期刊：
影响因子：
14
通讯作者：
Billiar, Kristen L.
中科院分区：
文献类型：
--
作者：
Kural, Mehmet H.;Billiar, Kristen L.
需要定量了解细胞、可溶性因子以及生物材料的生物和机械特性之间的复杂相互作用，以指导细胞重塑以再生健康组织而不是纤维收缩组织。在本研究中，我们表征了边界刚度和转化生长因子-β1 (TGF-β1) 对细胞产生的力和胶原蛋白积累的综合影响。我们首先通过在含有 TGF-β1 (0–5 ng/mL) 的化学成分确定的培养基中，在顺应性柱 (0.15–1.05 nN/nm) 之间的微型纤维蛋白凝胶内培养瓣膜间质细胞 (VIC)，生成细胞产生的张力响应这些因素的定量图。培养一周后，VIC 产生 100 至 3000 nN/细胞，多元回归模型首次证明了三维培养系统中这些因素之间的定量相互作用（协同作用）。然后，我们分离了微组织内张力的被动和主动成分，发现以高硬度和 TGF-β1 培养的细胞表达肌成纤维细胞标记物，并在基质中产生大量残余张力，但令人惊讶的是，它们无法响应膜去极化而产生额外的张力，这意味着持续最大收缩状态。相比之下，低硬度组和TGF-β1组中储存的残余张力可以忽略不计，表明释放时收缩的可能性较低。然后我们研究了 ECM 是否可以在低张力环境下产生，并发现 TGF-β1（而非 EGF）在低张力和高张力环境下大致相同地增加了从头胶原蛋白的积累。综合起来，这些发现表明等长细胞力、被动回缩和胶原蛋白生成可以通过独立改变边界刚度和 TGF-β1 浓度来调节。在不引起高主动张力的情况下刺激基质产生的能力将有助于开发坚固的组织工程心脏瓣膜和其他结缔组织替代品，其中最小化植入时的组织收缩至关重要。
A quantitative understanding of the complex interactions between cells, soluble factors, and the biological and mechanical properties of biomaterials is required to guide cell remodeling towards regeneration of healthy tissue rather than fibrocontractive tissue. In the present study, we characterized the combined effects of boundary stiffness and transforming growth factor-β1 (TGF-β1) on cell-generated forces and collagen accumulation. We first generated a quantitative map of cell-generated tension in response to these factors by culturing valvular interstitial cells (VICs) within micro-scale fibrin gels between compliant posts (0.15–1.05 nN/nm) in chemically-defined media with TGF-β1 (0–5 ng/mL). The VICs generated 100 to 3000 nN/cell after one week of culture, and multiple regression modeling demonstrated, for the first time, quantitative interaction (synergy) between these factors in a three-dimensional culture system. We then isolated passive and active components of tension within the micro-tissues and found that cells cultured with high levels of stiffness and TGF-β1 expressed myofibroblast markers and generated substantial residual tension in the matrix yet, surprisingly, were not able to generate additional tension in response to membrane depolarization signifying a state of continual maximal contraction. In contrast, negligible residual tension was stored in the low stiffness and TGF-β1 groups indicating a lower potential for shrinkage upon release. We then studied if ECM could be generated under the low tension environment and found that TGF-β1, but not EGF, increased de novo collagen accumulation in both low and high tension environments roughly equally. Combined, these findings suggest that isometric cell force, passive retraction, and collagen production can be tuned by independently altering boundary stiffness and TGF-β1 concentration. The ability to stimulate matrix production without inducing high active tension will aid in the development of robust tissue engineered heart valves and other connective tissue replacements where minimizing tissue shrinkage upon implantation is critical.
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影响因子：
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作者：
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