Development of artificial cornea utilizing with Apatite-polymer composites technique and nano-micro-fabrication technique

在本研究中，我们合成了可控制纳米结构的高分子-蛋白质复合材料用于光学部分，以及高分子-胶原-羟基磷灰石复合材料用于角膜假体的外围部分，为人工角膜的材料积累了基础数据。为了提高PVA水凝胶的生物相容性，我们开发了胶原、RGDS、IKVAV等生物分子对PVA水凝胶进行表面改性的技术。此外，我们还开发了新型的高相对分子质量聚苹果酸衍生物作为表面修饰剂和多糖和氨基酸的固定化技术作为人工配体来增强生物响应。为了实现人工角膜装置在宿主角膜上的牢固固定，我们开发了生物相容的PVA-羟基磷灰石复合材料。我们开发了两种不同的方法来制备PVA-羟基磷灰石复合材料，即交替浸渍法和颗粒捏合法。我们有…利用微细加工技术成功地将多孔结构引入到PVA-羟基磷灰石复合材料中。我们利用穿孔技术和电纺法制备的纳米纤维非织造布来构建三维结构，考察物理锚定效应是否有助于表皮的稳定。我们利用兔角膜原代细胞开始材料评价，并在此期间对兔角膜进行可行性研究。在PVA-胶原复合材料上证实了表皮的层次化，作为光学部分的材料。此外，还证实了细胞的黏附和铺展符合配体分子的密度，黏附的细胞在黏附蛋白固定的纳米纤维片上长期稳定。胶原固定化PVA水凝胶-羟基磷灰石复合材料对角膜细胞有明显的亲和力，表面拓扑结构对人工角膜上皮化和稳定性有很大影响，为长期使用的人工角膜设计提供了重要指标。通过将PVA-HAP复合材料植入兔角膜，观察到血管和宿主组织侵入到微孔结构中，这表明装置的牢固固定是可望实现的。较少

In this study, we synthesized the composites of the macromolecule - protein which controlled nano-structure to use for an optics part and composites such as macromolecule - collagen - hydoxyapatite complexes to use for a peripheral part of the corneal prosthesis, and accumulated the fundamental data as materials for artificial cornea. We developed surface modification technique of the PVA hydro-gel by the bio-molecules such as collagen, RGDS, IKVAV to improve the biocompatibility. And we also developed the new high molecular weight Poly(malic acid) derivatives as a surface modifier and polysaccharide and amino acids immobilization technique as a artificial ligand to enhance the biological responses.To achieve the strong fixation of the artificial corneal devices on the host cornea, we developed the biocompatible PVA-hydroxyapatite composites. We have developed two different methods, alternate soaking method and particles kneading method, to make the PVA-hydroxyapatite composites. We ha … More ve succeeded to introduce the porous structure into the PVA-hydroxyapatite composites by using micro-fabrication technique. We utilized the perforation technology and nanofiber nonwoven fabric prepared by the electrospinning methods to give the 3D structure and inspected whether an effect of physical anchoring contributed to stability of epidermis.We started the materials evaluation by using primary cells from rabbit cornea and the feasibility study on the rabbits cornea during the periods. Stratifications of epidermis were confirmed on the PVA-collagen composites as a material for optical part. In addition, the cell adhesion and spreading that agreed with density of a ligand molecule were confirmed.The adhered cells were stable for a long term on the adhesion protein immobilized nanofiber sheet. And clear affinity was given for the cornea cells on the collagen immobilization PVA hydro-gel-hydroxyapatite composites, The surface topology clearly influenced the epithelialization on an artificial cornea and the stabilization greatly, and a big indicator for a design of the artificial cornea which functioned for a long term was provided. As a result of implantation of PVA-HAp composites in the rabbit cornea, an invasion of a blood vessel and host tissues into the micropore structure were observed and the results suggested that the strong fixation of a device was realized was expected. Less