Transforming growth factor-β3 regulates assembly of a non-fibrotic matrix in a 3D corneal model.
Transforming growth factor-β3 regulates assembly of a non-fibrotic matrix in a 3D corneal model.
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DOI：
10.1002/term.429
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发表时间：
2011-08
影响因子：
3.3
通讯作者：
Zieske, J. D.
中科院分区：
文献类型：
--
作者：
Karamichos, D.;Hutcheon, A. E. K.;Zieske, J. D.
多年来，角膜组织工程引起了许多研究人员的关注，部分原因是角膜的无血管性和相对直接的结构。然而，这种光学透明组织的高度组织性和结构化性质提出了巨大的挑战。我们之前开发了一种模型，其中稳定的维生素 C (VitC) 衍生物刺激人角膜成纤维细胞 (HCF) 自组装细胞外基质 (ECM)。添加TGF-β1增强了ECM的组装；然而，它伴随着特定纤维化标志物的上调。在这项研究中，我们测试了所有三种 TGF-β 亚型（-β1、-β2 和 -β3）对 ECM 产生以及纤维化标志物表达的影响。 HCF 在四种培养基条件下生长 4 周：（对照）仅 VitC； (T1)维生素C+TGF-β1； (T2)维生素C+TGF-β2；或 (T3) VitC+TGF-β3。使用蛋白质印迹、TEM 和间接免疫荧光 (IF) 分析培养物。与对照相比，所有 TGF-β 同工型均能刺激基质产生约 3 倍。 IF显示T1和T2中存在III型胶原和平滑肌肌动蛋白（SMA）；然而，T3 几乎没有表达。在蛋白质印迹中，与其他条件相比，T3 刺激的 III 型/I 型胶原比例较低。此外，TEM 表明 T3 刺激了更高水平的基质排列和组织。受 VitC 和 TGF-β3 刺激的 HCF 似乎会生成模仿正常成人或发育中的人类角膜的基质；然而，TGF-β1 和-β2 正在驱动构建体走向更加纤维化的路径。
Corneal tissue engineering has attracted the attention of many researchers over the years in part due to the cornea’s avascularity and relatively, straight forward structure. However, the highly organized and structured nature of this optically clear tissue has presented a great challenge. We have previously developed a model where human corneal fibroblasts (HCFs) are stimulated by a stable Vitamin C (VitC) derivative to self-assemble an extracellular matrix (ECM). Addition of TGF-β1 enhanced the assembly of ECM; however, it was accompanied by the upregulation of specific fibrotic markers. In this study, we tested the effects of all three TGF-β isoforms (-β1, -β2 and -β3) on ECM production, as well as, expression of fibrotic markers. HCFs were grown in four media conditions for 4 weeks: (Control) VitC only; (T1) VitC+TGF-β1; (T2) VitC+TGF-β2; or (T3) VitC+TGF-β3. Cultures were analyzed with Western Blots, TEM and indirect-immunofluorescence (IF). Compared to controls, all TGF-β isoforms stimulated matrix production by ~3 times. IF showed the presence of type III collagen and smooth muscle actin (SMA) in T1 and T2; however, T3 showed little, to no, expression. In western blots, T3 stimulated a lower type III/type I collagen ratio when compared to the other conditions. In addition, TEM indicated that T3 stimulated a higher level of matrix alignment and organization. HCFs stimulated by VitC and TGF-β3 appear to generate a matrix that mimics the normal adult or developing human cornea; whereas, TGF-β1 and -β2 are driving the constructs toward a more fibrotic path.
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影响因子：
3.4
作者：
Jester, JV;Huang, JY;Cavanagh, HD
通讯作者：
Cavanagh, HD
影响因子：
4.4
作者：
Karamichos, Dimitris;Guo, Xiaoqing Q.;Zieske, James D.
通讯作者：
Zieske, James D.
影响因子：
4.8
作者：
Long, CJ;Roth, MR;Funderburgh, JL
通讯作者：
Funderburgh, JL
影响因子：
3.3
作者：
Gonzalez-Andrades, M.;Garzon, I.;Alaminos, M.
通讯作者：
Alaminos, M.
影响因子：
4.4
作者：
Guo, Xiaoqing;Hutcheon, Audrey E. K.;Ruberti, Jeffrey W.
通讯作者：
Ruberti, Jeffrey W.