Recreating the Trabecular Outflow Tissue on Implantable, Micropatterned, Ultrathin, Porous Polycaprolactone Scaffolds.
Recreating the Trabecular Outflow Tissue on Implantable, Micropatterned, Ultrathin, Porous Polycaprolactone Scaffolds.
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DOI：
10.3390/bioengineering10060679
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发表时间：
2023-06-02
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--
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青光眼，其中眼内压（IOP）升高导致视神经损伤和视力丧失，是全世界不可逆失明的最主要原因之一。在原发性开角型青光眼中，增加的IOP是功能障碍的人小梁网（HTM）细胞不能适当地调节眼房水流出的结果。青光眼的一种潜在的未来治疗方法是用组织工程替代品取代受损的HTM细胞，从而恢复适当的液体流出。聚己内酯（PCL）是一种多功能、可生物降解和可植入的材料，广泛用于细胞培养和组织工程。在这项工作中，使用牺牲工艺光刻制造PCL支架，以产生具有特定尺寸和形状的开口的亚微米厚支架（例如，网格、六边形图案）。通过扫描电镜、四氮唑代谢活性测定和F-肌动蛋白的细胞骨架组织来评估HTM细胞在明胶涂覆的PCL支架上的生长。通过免疫细胞化学和qPCR分析评估HTM特异性标志物的表达和ECM沉积。具有网格图案的明胶涂覆的微图案化的多孔PCL支架支持适当的HTM细胞生长、细胞骨架组织、HTM标记物表达和ECM沉积，证明了使用这些PCL支架来组织工程化可植入的健康眼流出组织的可行性。
Glaucoma, where increased intraocular pressure (IOP) leads to damage to the optic nerve and loss of sight, is amongst the foremost causes of irreversible blindness worldwide. In primary open angle glaucoma, the increased IOP is a result of the malfunctioning human trabecular meshwork (HTM) cells’ inability to properly regulate the outflow of aqueous humor from the eye. A potential future treatment for glaucoma is to replace damaged HTM cells with a tissue-engineered substitute, thus restoring proper fluid outflow. Polycaprolactone (PCL) is a versatile, biodegradable, and implantable material that is widely used for cell culture and tissue engineering. In this work, PCL scaffolds were lithographically fabricated using a sacrificial process to produce submicron-thick scaffolds with openings of specific sizes and shapes (e.g., grid, hexagonal pattern). The HTM cell growth on gelatin-coated PCL scaffolds was assessed by scanning electron microscopy, tetrazolium metabolic activity assay, and cytoskeletal organization of F-actin. Expression of HTM-specific markers and ECM deposition were assessed by immunocytochemistry and qPCR analysis. Gelatin-coated, micropatterned, ultrathin, porous PCL scaffolds with a grid pattern supported proper HTM cell growth, cytoskeleton organization, HTM-marker expression, and ECM deposition, demonstrating the feasibility of using these PCL scaffolds to tissue-engineer implantable, healthy ocular outflow tissue.
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DOI：
10.1136/bjophthalmol-2015-307223
发表时间：
2016-01
期刊：
The British journal of ophthalmology
影响因子：
--
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通讯作者：
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