Evaluation of silk biomaterials in combination with extracellular matrix coatings for bladder tissue engineering with primary and pluripotent cells.
Evaluation of silk biomaterials in combination with extracellular matrix coatings for bladder tissue engineering with primary and pluripotent cells.
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DOI：
10.1371/journal.pone.0056237
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Mauney JR
中科院分区：
文献类型：
--
作者：
Franck D;Gil ES;Adam RM;Kaplan DL;Chung YG;Estrada CR Jr;Mauney JR
丝基生物材料与细胞外基质（ECM）涂层的组合被评估为细胞接种膀胱组织工程方法的模板。通过凝胶纺丝工艺生产两组结构不同的丝支架，其由光滑、致密的多层片（组1）或粗糙、多孔的层状片（组2）组成。单独的或用I型或IV型胶原或纤连蛋白包被的支架独立地评估其支持原代细胞系（包括人膀胱平滑肌细胞（SMC）和尿路上皮细胞）以及多能细胞群（如鼠胚胎干细胞（ESC）和诱导多能干（iPS）细胞）的附着、增殖和分化的能力。AlamarBlue评估显示，与未涂覆的第2组对照和所有第1组支架变体相比，纤连蛋白涂覆的第2组支架促进了最高程度的原代SMC和尿路上皮细胞附着。真实的时间RT-PCR和免疫组化（IHC）分析表明，两个纤连蛋白包被的丝组都允许SMC收缩分化，这通过TGFβ1刺激后α-肌动蛋白和SM 22 α mRNA和蛋白表达水平的显著上调来确定。在IHC分析后，在对照组和纤连蛋白包被组培养的尿路上皮细胞中观察到上皮分化标志物细胞角蛋白的显著表达。通过alamarBlue对丝基质用于ESC和iPS细胞附着的评估显示，与所有其他支架制剂相比，纤连蛋白包被的第2组支架促进了最高水平。此外，真实的时间RT-PCR和IHC分析表明，纤连蛋白包被的第2组支架促进ESC和iPS细胞向尿路上皮和平滑肌谱系分化，以响应全反式视黄酸，如通过诱导尿斑蛋白和收缩基因和蛋白质表达所评估的。这些结果表明，丝支架支持与膀胱组织工程相关的原代和多能细胞反应，并且支架形态和纤连蛋白涂层影响这些过程。
Silk-based biomaterials in combination with extracellular matrix (ECM) coatings were assessed as templates for cell-seeded bladder tissue engineering approaches. Two structurally diverse groups of silk scaffolds were produced by a gel spinning process and consisted of either smooth, compact multi-laminates (Group 1) or rough, porous lamellar-like sheets (Group 2). Scaffolds alone or coated with collagen types I or IV or fibronectin were assessed independently for their ability to support attachment, proliferation, and differentiation of primary cell lines including human bladder smooth muscle cells (SMC) and urothelial cells as well as pluripotent cell populations, such as murine embryonic stem cells (ESC) and induced pluripotent stem (iPS) cells. AlamarBlue evaluations revealed that fibronectin-coated Group 2 scaffolds promoted the highest degree of primary SMC and urothelial cell attachment in comparison to uncoated Group 2 controls and all Group 1 scaffold variants. Real time RT-PCR and immunohistochemical (IHC) analyses demonstrated that both fibronectin-coated silk groups were permissive for SMC contractile differentiation as determined by significant upregulation of α-actin and SM22α mRNA and protein expression levels following TGFβ1 stimulation. Prominent expression of epithelial differentiation markers, cytokeratins, was observed in urothelial cells cultured on both control and fibronectin-coated groups following IHC analysis. Evaluation of silk matrices for ESC and iPS cell attachment by alamarBlue showed that fibronectin-coated Group 2 scaffolds promoted the highest levels in comparison to all other scaffold formulations. In addition, real time RT-PCR and IHC analyses showed that fibronectin-coated Group 2 scaffolds facilitated ESC and iPS cell differentiation toward both urothelial and smooth muscle lineages in response to all trans retinoic acid as assessed by induction of uroplakin and contractile gene and protein expression. These results demonstrate that silk scaffolds support primary and pluripotent cell responses pertinent to bladder tissue engineering and that scaffold morphology and fibronectin coatings influence these processes.
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影响因子：
14
作者：
Brown, AL;Farhat, W;Woodhouse, KA
通讯作者：
Woodhouse, KA
影响因子：
5.2
作者：
Metallo, Christian M.;Ji, Lin;Palecek, Sean P.
通讯作者：
Palecek, Sean P.
影响因子：
8.3
作者：
Kaimoto, Taeko;Yasuda, Osamu;Rakugi, Hiromi
通讯作者：
Rakugi, Hiromi
影响因子：
3.5
作者：
Chun, Young Wook;Khang, Dongwoo;Webster, Thomas J.
通讯作者：
Webster, Thomas J.
DOI：
10.1385/scr:1:2:099
发表时间：
2005-01-01
期刊：
STEM CELL REVIEWS
影响因子：
--
作者：
Hwang, Woo Suk;Lee, Byeong Chun;Kang, Sung Keun
通讯作者：
Kang, Sung Keun