Open AccessModified insulin-like growth factor 1 containing collagen-binding domain for nerve regeneration
Author(s) -
Jianan Li,
Cheng Zhao,
Shaojun Li,
Jun Zhang,
Zhenhua Li,
Zhengxue Qiao,
Xiaoyu Yang,
Chunfang Zan
Publication year - 2018
Publication title -
neural regeneration research/neural regeneration research
Language(s) - Uncategorized
Resource type - Journals
SCImago Journal Rank - 0.93
H-Index - 38
eISSN - 1876-7958
pISSN - 1673-5374
DOI - 10.4103/1673-5374.226400
Subject(s) - nerve growth factor , recombinant dna , growth factor , neurofilament , regeneration (biology) , schwann cell , transfection , insulin like growth factor , microbiology and biotechnology , receptor , chemistry , biology , medicine , endocrinology , biochemistry , immunohistochemistry , gene
Insulin-like growth factor 1 (IGF-1) is a potential nutrient for nerve repair. However, it is impractical as a therapy because of its limited half-life, rapid clearance, and limited target specificity. To achieve targeted and long-lasting treatment, we investigated the addition of a binding structure by fusing a collagen-binding domain to IGF-1. After confirming its affinity for collagen, the biological activity of this construct was examined by measuring cell proliferation after transfection into PC12 and Schwann cells using a 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay. Immunofluorescence staining was conducted to detect neurofilament and microtubule-associated protein 2 expression, while real time-polymerase chain reaction was utilized to determine IGF-1 receptor and nerve growth factor mRNA expression. Our results demonstrate a significant increase in collagen-binding activity of the recombinant protein compared with IGF-1. Moreover, the recombinant protein promoted proliferation of PC12 and Schwann cells, and increased the expression of neurofilament and microtubule-associated protein 2. Importantly, the recombinant protein also stimulated sustained expression of IGF-1 receptor and nerve growth factor mRNA for days. These results show that the recombinant protein achieved the goal of targeting and long-lasting treatment, and thus could become a clinically used factor for promoting nerve regeneration with a prolonged therapeutic effect.