Layer-by-layer immobilized catalase on electrospun nanofibrous mats protects against oxidative stress induced by hydrogen peroxide.

Rong Huang, Hongbing Deng, Tongjian Cai, Yingfei Zhan, Xiankai Wang, Xuanxuan Chen, Ailing Ji, Xueyong Lil

Index: J. Biomed. Nanotechnol. 10(7) , 1346-58, (2014)

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Abstract

Catalase, a kind of redox enzyme and generally recognized as an efficient agent for protecting cells against hydrogen peroxide (H2O2)-induced cytotoxicity. The immobilization of catalase was accomplished by depositing the positively charged chitosan and the negatively charged catalase on electrospun cellulose nanofibrous mats through electrospining and layer-by-layer (LBL) techniques. The morphology obtained from Field emission scanning electron microscopy (FE-SEM) indicated that more orderly arranged three-dimension (3D) structure and roughness formed with increasing the number of coating bilayers. Besides, the enzyme-immobilized nanofibrous mats were found with high enzyme loading and activity, moreover, X-ray photoelectron spectroscopy (XPS) results further demonstrated the successful immobilization of chitosan and catalase on cellulose nanofibers support. Furthermore, we evaluated the cytotoxicity induced by hydrogen peroxide in the Human umbilical vascular endothelial cells with or without pretreatment of nanofibrous mats by MTT assay, LDH activity and Flow cytometric evaluation, and confirmed the pronounced hydrogen peroxide-induced toxicity, but pretreatment of immobilized catalase reduced the cytotoxicity and protected cells against hydrogen peroxide-induced cytotoxic effects which were further demonstrated by scanning electron microscopy (SEM) and Transmission Electron Microscopy (TEM) images. The data pointed toward a role of catalase-immobilized nanofibrous mats in protecting cells against hydrogen peroxide-induced cellular damage and their potential application in biomedical field.