Quantifying Cell-Derived Changes in Collagen Synthesis, Alignment, and Mechanics in a 3D Connective Tissue Model.
Quantifying Cell-Derived Changes in Collagen Synthesis, Alignment, and Mechanics in a 3D Connective Tissue Model.
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DOI：
10.1002/advs.202103939
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发表时间：
2022-04
期刊：
影响因子：
--
通讯作者：
Morgan JR
中科院分区：
文献类型：
--
作者：
Wilks BT;Evans EB;Howes A;Hopkins CM;Nakhla MN;Williams G;Morgan JR
细胞外基质（ECM）合成、组织和力学的失调是纤维化和癌症等疾病的标志性特征。然而，大多数体外模型无法概括富含胶原蛋白组织的三维（3D）多尺度分层结构，因此无法反映天然或疾病表型。在本文中，使用接种到定制制造的3D非粘性琼脂糖模具中的原代人成纤维细胞，提出了一种新的策略来指导工程化3D环形组织结构的形态发生，其具有概括纤维结缔组织的关键特征的拉伸和组织学特性。为了表征从单分散细胞到高度排列的富含胶原的基质的转变，采用了整合组织学、多光子二次谐波发生和电子显微镜的多模式方法。然后将胶原合成和排列中的结构变化映射到组织力学和总胶原含量中的功能差异。由于不存在外源添加的支架材料，该模型能够直接定量响应于添加或去除相关生物分子扰动的3D基质合成、对齐和力学中的细胞衍生变化。为了说明这一点，定量了营养成分、胎牛血清、rho激酶抑制剂以及促纤维化和抗纤维化化合物对ECM合成、3D胶原结构和机械表型的影响。提出了一种三维（3D）无支架人结缔组织模型，该模型有助于高度对齐、富含胶原蛋白的细胞外基质（ECM）的从头合成。该模型能够直接量化3D ECM合成，对齐和力学响应生物分子扰动的细胞衍生变化，并可能有助于更好地理解ECM的复杂疾病，如纤维化。
Dysregulation of extracellular matrix (ECM) synthesis, organization, and mechanics are hallmark features of diseases like fibrosis and cancer. However, most in vitro models fail to recapitulate the three‐dimensional (3D) multi‐scale hierarchical architecture of collagen‐rich tissues and as a result, are unable to mirror native or disease phenotypes. Herein, using primary human fibroblasts seeded into custom fabricated 3D non‐adhesive agarose molds, a novel strategy is proposed to direct the morphogenesis of engineered 3D ring‐shaped tissue constructs with tensile and histological properties that recapitulate key features of fibrous connective tissue. To characterize the shift from monodispersed cells to a highly‐aligned, collagen‐rich matrix, a multi‐modal approach integrating histology, multiphoton second‐harmonic generation, and electron microscopy is employed. Structural changes in collagen synthesis and alignment are then mapped to functional differences in tissue mechanics and total collagen content. Due to the absence of an exogenously added scaffolding material, this model enables the direct quantification of cell‐derived changes in 3D matrix synthesis, alignment, and mechanics in response to the addition or removal of relevant biomolecular perturbations. To illustrate this, the effects of nutrient composition, fetal bovine serum, rho‐kinase inhibitor, and pro‐ and anti‐fibrotic compounds on ECM synthesis, 3D collagen architecture, and mechanophenotype are quantified. A three‐dimensional (3D) scaffold‐free human connective tissue model is presented that facilitates the de novo synthesis of a highly‐aligned, collagen‐rich extracellular matrix (ECM). This model enables the direct quantification of cell‐derived changes in 3D ECM synthesis, alignment, and mechanics in response to biomolecular perturbations and may be useful for better understanding complex diseases of the ECM like fibrosis.
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DOI：
10.1073/pnas.1003743107
发表时间：
2010-07-13
影响因子：
11.1
作者：
Manning, M. Lisa;Foty, Ramsey A.;Schoetz, Eva-Maria
通讯作者：
Schoetz, Eva-Maria
影响因子：
6.9
作者：
Kalson, Nicholas S.;Holmes, David F.;Kapacee, Zoher;Otermin, Iker;Lu, Yinhui;Ennos, Roland A.;Canty-Laird, Elizabeth G.;Kadler, Karl E.
通讯作者：
Kadler, Karl E.
影响因子：
64.8
作者：
Harris CR;Millman KJ;van der Walt SJ;Gommers R;Virtanen P;Cournapeau D;Wieser E;Taylor J;Berg S;Smith NJ;Kern R;Picus M;Hoyer S;van Kerkwijk MH;Brett M;Haldane A;Del Río JF;Wiebe M;Peterson P;Gérard-Marchant P;Sheppard K;Reddy T;Weckesser W;Abbasi H;Gohlke C;Oliphant TE
通讯作者：
Oliphant TE
影响因子：
9.8
作者：
Coller HA;Sang L;Roberts JM
通讯作者：
Roberts JM
影响因子：
4.1
作者：
Livoti, Christine M.;Morgan, Jeffrey R.
通讯作者：
Morgan, Jeffrey R.