Original Article
Copyright ©2011 Baishideng Publishing Group Co., Limited. All rights reserved.
World J Gastroenterol. Jun 14, 2011; 17(22): 2774-2780
Published online Jun 14, 2011. doi: 10.3748/wjg.v17.i22.2774
Influence of chitosan nanofiber scaffold on porcine endogenous retroviral expression and infectivity in pig hepatocytes
Bing Han, Xiao-Lei Shi, Jiang-Qiang Xiao, Yue Zhang, Xue-Hui Chu, Jin-Yang Gu, Jia-Jun Tan, Zhong-Ze Gu, Yi-Tao Ding
Bing Han, Xiao-Lei Shi, Jiang-Qiang Xiao, Xue-Hui Chu, Jin-Yang Gu, Jia-Jun Tan, Yi-Tao Ding, Department of Hepatobiliary Surgery, the Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing 210008, Jiangsu Province, China
Yue Zhang, Yi-Tao Ding, Department of Hepatobiliary Surgery, Drum Tower Clinical Medical College of Nanjing Medical University, Nanjing 210008, Jiangsu Province, China
Zhong-Ze Gu, State Key Laboratory of Bioelectronics, Southeast University, Nanjing 210000, Jiangsu Province, China
Author contributions: Han B and Shi XL contributed equally to this work; Han B and Shi XL designed the research; Han B, Xiao JQ, Zhang Y, Chu XH, Gu JY and Tan JJ performed the research; Gu ZZ and Ding YT contributed new reagents/analytic tools; Han B and Shi XL analyzed the data and wrote the paper.
Supported by The Natural Science Foundation of Jiangsu Province, No. BK2006008; the foundation of Medical Center of Jiangsu Province, No.ZX200605
Correspondence to: Yi-Tao Ding, Professor, Department of Hepatobiliary Surgery, the Affiliated Drum Tower Hospital of Nanjing University Medical School, No. 321 Zhongshan Road, Nanjing 210008, Jiangsu Province, China. yitaoding@hotmail.com
Telephone: +86-25-83304616 Fax: +86-25-83317016
Received: September 15, 2010
Revised: November 15, 2010
Accepted: November 22, 2010
Published online: June 14, 2011
Abstract

AIM: To investigate the influence of chitosan nanofiber scaffold on the production and infectivity of porcine endogenous retrovirus (PERV) expressed by porcine hepatocytes.

METHODS: Freshly isolated porcine hepatocytes were cultured with or without chitosan nanofiber scaffold (defined as Nano group and Hep group) for 7 d. The daily collection of culture medium was used to detect reverse transcriptase (RT) activity with RT activity assay kits and PERV RNA by reverse transcription-polymerase chain reaction (PCR) and real time PCR with the PERV specific primers. And Western blotting was performed with the lysates of daily retrieved cells to determine the PERV protein gag p30. Besides, the in-vitro infectivity of the supernatant was tested by incubating the human embryo kidney 293 (HEK293) cells.

RESULTS: The similar changing trends between two groups were observed in real time PCR, RT activity assay and Western blotting. Two peaks of PERV expression at 10H and Day 2 were found and followed by a regular decline. No significant difference was found between two groups except the significantly high level of PERV RNA at Day 6 and PERV protein at Day 5 in Nano group than that in Hep group. And in the in-vitro infection experiment, no HEK293 cell was infected by the supernatant.

CONCLUSION: Chitosan nanofiber scaffold might prolong the PERV secreting time in pig hepatocytes but would not obviously influence its productive amount and infectivity, so it could be applied in the bioartificial liver without the increased risk of the virus transmission.

Keywords: Chitosan nanofiber scaffold; Porcine hepatocyte; Porcine endogenous retrovirus; Bioartificial liver