Lai Jui-Yang, Ma David Hui-Kang
Institute of Biochemical and Biomedical Engineering, Chang Gung University, Taoyuan 33302, Taiwan, Republic of China; Biomedical Engineering Research Center, Chang Gung University, Taoyuan 33302, Taiwan, Republic of China; Center for Tissue Engineering, Chang Gung Memorial Hospital, Taoyuan 33305, Taiwan, Republic of China; Department of Ophthalmology, Chang Gung Memorial Hospital, Taoyuan 33305, Taiwan, Republic of China; Department of Materials Engineering, Ming Chi University of Technology, New Taipei City 24301, Taiwan, Republic of China.
Center for Tissue Engineering, Chang Gung Memorial Hospital, Taoyuan 33305, Taiwan, Republic of China; Department of Ophthalmology, Chang Gung Memorial Hospital, Taoyuan 33305, Taiwan, Republic of China; Department of Chinese Medicine, Chang Gung University, Taoyuan 33302, Taiwan, Republic of China.
Mater Sci Eng C Mater Biol Appl. 2017 Mar 1;72:150-159. doi: 10.1016/j.msec.2016.11.067. Epub 2016 Nov 18.
Given that the presence of aldehyde groups on the oxidized sugar residues may pose toxicity concerns, it is necessary to examine the safety of gelatin microcarriers (GMC) functionalized with oxidized hyaluronic acid (oHA) for potential ophthalmic applications. In this study, the ocular biocompatibility of biopolymer microcarriers was investigated in vitro using primary rabbit corneal cell cultures and in vivo using the anterior chamber of the rabbit eye model. Our results showed that different types of corneal cells including epithelial, stromal, and endothelial cells remain viable and actively proliferate following 2 and 4days of exposure to test materials. In addition, similar interleukin-6 gene expression levels and comet tail lengths were seen in the presence and absence of biopolymer microcarriers, suggesting no cellular inflammation and genotoxicity. After 7 and 14days of intracameral injection in the rabbit eyes, both the GMC samples and their counterparts functionalized with oHA were well tolerated in the ocular anterior chamber as demonstrated by slit-lamp biomicroscopy. Clinical observations including specular microscopic examinations, corneal topography, and corneal thickness measurements also showed that the rabbits bearing biopolymer microcarriers exhibit no signs of corneal edema and astigmatism as well as endothelial damage, indicating the absence of tissue response. It is concluded that the GMC materials functionalized with oHA (oxidation level: 10.4±0.9%) are compatible toward corneal cells and ocular anterior segment tissues at a concentration of 10mg/ml. The information about the effect of coupling of aldehyde-functionalized HA to gelatin on in vitro and in vivo biocompatibility of biopolymer composites can be used as further development of corneal stromal cell microcarriers for tissue engineering applications.
鉴于氧化糖残基上醛基的存在可能引发毒性问题，有必要研究用氧化透明质酸（oHA）功能化的明胶微载体（GMC）在潜在眼科应用中的安全性。在本研究中，使用原代兔角膜细胞培养物在体外以及使用兔眼前房模型在体内研究了生物聚合物微载体的眼生物相容性。我们的结果表明，包括上皮细胞、基质细胞和内皮细胞在内的不同类型角膜细胞在接触测试材料2天和4天后仍保持活力并积极增殖。此外，在有和没有生物聚合物微载体的情况下，白细胞介素-6基因表达水平和彗星尾长度相似，表明没有细胞炎症和基因毒性。在兔眼进行前房注射7天和14天后，裂隙灯生物显微镜检查表明，GMC样品及其用oHA功能化的对应物在前房中耐受性良好。包括镜面显微镜检查、角膜地形图和角膜厚度测量在内的临床观察还表明，携带生物聚合物微载体的兔子没有角膜水肿、散光以及内皮损伤的迹象，表明没有组织反应。得出的结论是，用oHA功能化的GMC材料（氧化水平：10.4±0.9%）在浓度为10mg/ml时对角膜细胞和眼前节组织具有相容性。关于醛基功能化的HA与明胶偶联对生物聚合物复合材料体外和体内生物相容性的影响的信息可用于进一步开发用于组织工程应用的角膜基质细胞微载体。
