Mesh-supported submicron parylene-C membranes for culturing retinal pigment epithelial cells
Mesh-supported submicron parylene-C membranes for culturing retinal pigment epithelial cells
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DOI：
10.1007/s10544-012-9645-8
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发表时间：
2012-08-01
影响因子：
2.8
通讯作者：
Tai, Yu-Chong
中科院分区：
文献类型：
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作者：
Lu, Bo;Zhu, Danhong;Tai, Yu-Chong
在这项工作中，提出了一种网状支撑的亚微米聚对二甲苯-C膜（MSPM）作为人工布鲁赫膜用于治疗年龄相关性黄斑变性（AMD）。任何人造布鲁赫膜必须首先满足两个重要要求。首先，它应该像健康人的布鲁赫膜一样具有渗透性，以支持营养物质的运输。其次，它应该能够支持具有体内样形态和功能的视网膜色素上皮（RPE）细胞的粘附和增殖。尽管聚对二甲苯-C在许多生物医学应用中被广泛用作阻挡层，但发现具有亚微米厚度的聚对二甲苯-C膜对大分子是半渗透的。我们首先测量亚微米聚对二甲苯-C的渗透性，发现0.15-0.30 μ m聚对二甲苯-C具有与健康人布鲁赫膜相似的渗透性。盲孔灌注细胞活力实验进一步证明，营养物质和大分子可以扩散通过0.30 μ m聚对二甲苯-C来滋养细胞。设计了一种网格支撑的亚微米聚对二甲苯-C膜（MSPM）结构，以提高基底的机械强度。在MSPM（含0.30 μ mParylene-C）上进行的体外细胞培养显示，H9-RPE细胞能够粘附、增殖、形成具有紧密细胞内连接的上皮单层，并且变得具有良好极化的微绒毛，其表现出与体内RPE细胞相似的特征。这些研究已经证明了MSPM作为人工Bruch膜用于RPE细胞移植的潜力。
In this work, a mesh-supported submicron parylene-C membrane (MSPM) is proposed as an artificial Bruch's membrane for the therapy of age-related macular degeneration (AMD). Any artificial Bruch's membrane must first satisfy two important requirements. First, it should be as permeable as healthy human Bruch's membrane to support nutrients transportation. Secondly, it should be able to support the adherence and proliferation of retinal pigment epithelial (RPE) cells with in vivo-like morphologies and functions. Although parylene-C is widely used as a barrier layer in many biomedical applications, it is found that parylene-C membranes with submicron thickness are semipermeable to macromolecules. We first measure the permeability of submicron parylene-C and find that 0.15-0.30 mu m parylene-C has similar permeability to healthy human Bruch's membranes. Blind-well perfusion cell viability experiments further demonstrate that nutrients and macromolecules can diffuse across 0.30 mu m parylene-C to nourish the cells. A mesh-supported submicron parylene-C membrane (MSPM) structure is design to enhance the mechanical strength of the substrate. In vitro cells culture on the MSPM (with 0.30 mu m ultrathin parylene-C) shows that H9-RPE cells are able to adhere, proliferate, form epithelial monolayer with tight intracellular junctions, and become well-polarized with microvilli, which exhibit similar characteristics to RPE cells in vivo. These studies have demonstrated the potential of the MSPM as an artificial Bruch's membrane for RPE cell transplantation.