Shao Yi, Yu Yao, Pei Chong-Gang, Zhou Qiong, Liu Qiu-Ping, Tan Gang, Li Jing-Ming, Gao Gui-Ping, Yang Lu
Department of Ophthalmology, the First Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi Province, China.
Int J Ophthalmol. 2012;5(4):415-8. doi: 10.3980/j.issn.2222-3959.2012.04.02. Epub 2012 Aug 18.
To develop a new decellularization method depended upon the natural corneal structure and to harvest an ideal scaffold with good biocompatibilities for corneal reconstruction.
The acellular cornea matrix (ACM) were prepared from de-epithelium fresh porcine corneas (DFPCs) by incubation with 100% fresh human sera and additional electrophoresis at 4°C. Human corneal epithelial cells (HCEs) were used for the cytotoxicity tests of ACM. ACM were implanted into the Enhanced Green Fluorecence Protein (eGFP) transgenic mouse anterior chamber for evaluation of histocompatibility.
HE and GSIB4 results showed fresh porcine cornea matrix with 100% human sera and electrophoresis could entirely decellularize stromal cell without reducing its transparency. ACM had no cytotoxic effect ex vivo. Animal test showed there was no rejection for one month after surgery.
These results provide a decellularizing approach for the study of corneal tissue engineering and had the broader implications for the field of biological tissue engineering in other engineered organ or tissue matrix.
开发一种依赖天然角膜结构的新型去细胞化方法,并获取具有良好生物相容性的理想支架用于角膜重建。
通过与100%新鲜人血清孵育并在4℃下进行额外电泳,从去上皮新鲜猪角膜(DFPCs)制备脱细胞角膜基质(ACM)。用人角膜上皮细胞(HCEs)进行ACM的细胞毒性测试。将ACM植入增强型绿色荧光蛋白(eGFP)转基因小鼠前房以评估组织相容性。
苏木精-伊红染色(HE)和GSIB4结果显示,用100%人血清和电泳处理的新鲜猪角膜基质可使基质细胞完全去细胞化,且不降低其透明度。ACM在体外无细胞毒性作用。动物试验表明,术后1个月无排斥反应。
这些结果为角膜组织工程研究提供了一种去细胞化方法,对其他工程化器官或组织基质的生物组织工程领域具有更广泛的意义。