Gene therapy for nucleus pulposus regeneration by heme oxygenase-1 plasmid DNA carried by mixed polyplex micelles with thermo-responsive heterogeneous coronas

Ganjun Feng; Hongying Chen; Junjie Li; Qiang Huang; Melanie J Gupte; Hao Liu; Yueming Song; Zhishen Ge

doi:10.1016/j.biomaterials.2015.02.024

Gene therapy for nucleus pulposus regeneration by heme oxygenase-1 plasmid DNA carried by mixed polyplex micelles with thermo-responsive heterogeneous coronas

Biomaterials. 2015 Jun:52:1-13. doi: 10.1016/j.biomaterials.2015.02.024. Epub 2015 Feb 18.

Authors

Ganjun Feng¹, Hongying Chen², Junjie Li³, Qiang Huang², Melanie J Gupte⁴, Hao Liu¹, Yueming Song¹, Zhishen Ge⁵

Affiliations

¹ Department of Orthopedic Surgery, West China Hospital, Sichuan University, Chengdu 610041, China.
² Technology Center for Public Research, West China Hospital, Sichuan University, Chengdu 610041, China.
³ CAS Key Laboratory of Soft Matter Chemistry, Department of Polymer Science and Engineering, University of Science and Technology of China, Hefei 230026, China.
⁴ Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109, USA.
⁵ CAS Key Laboratory of Soft Matter Chemistry, Department of Polymer Science and Engineering, University of Science and Technology of China, Hefei 230026, China. Electronic address: gezs@ustc.edu.cn.

PMID: 25818409
DOI: 10.1016/j.biomaterials.2015.02.024

Abstract

Safe and high-efficiency gene therapy for nucleus pulposus (NP) regeneration was urgently desired to treat disc degeneration-associated diseases. In this work, an efficient nonviral cationic block copolymer gene delivery system was used to deliver therapeutic plasmid DNA (pDNA), which was prepared via complexation between the mixed cationic block copolymers, poly(ethylene glycol)-block-poly{N-[N-(2-aminoethyl)-2-aminoehtyl]aspartamide} [PEG-b-PAsp(DET)] and poly(N-isopropylacrylamide)-block-PAsp(DET) [PNIPAM-b-PAsp(DET)], and pDNA at 25 °C. The mixed polyplex micelles (MPMs) containing heterogeneous coronas with hydrophobic and hydrophilic microdomains coexisting could be obtained upon heating from 25 to 37 °C, which showed high tolerability against nuclease and strong resistance towards protein adsorption. The gene transfection efficiency of MPMs in NP cells was significantly higher than that of regular polyplex micelles prepared from sole block copolymer of PEG-b-PAsp(DET) (SPMs) in in vitro and in vivo evaluation due to the synergistic effect of improved colloidal stability and low cytotoxicity. High expression of heme oxygenase-1 (HO-1) in NP cells transfected by MPMs loading HO-1 pDNA significantly decreased the expression activity of matrix metalloproteinases 3 (MMP-3) and cyclo-oxygenase-2 (COX-2) induced by interleukin-1β (IL-1β), and simultaneously increased the NP phenotype-associated genes such as aggrecan, type II collagen, and SOX-9. Moreover, the therapeutic effects of MPMs loading pDNA were tested to treat disc degeneration induced by stab injury. The results demonstrated that administration of HO-1 pDNA carried by MPMs in rat tail discs apparently reduced inflammatory responses induced by need stab and increased glycosaminoglycan (GAG) content, finally achieving better therapeutic efficacy as compared with SPMs. Consequently, MPMs loading HO-1 pDNA were demonstrated to be potential as a safe and high-efficiency nonviral gene delivery system for retarding or regenerating the degenerative discs.

Keywords: Gene therapy; Nonviral gene carrier; Nucleus pulposus regeneration; Polyplex micelles; Thermo-responsive.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Acrylic Resins / chemistry
Animals
Cells, Cultured
Cervical Vertebrae / metabolism
Cervical Vertebrae / physiology*
DNA / administration & dosage*
DNA / genetics
DNA / therapeutic use
Gene Expression Regulation
Genetic Therapy / methods*
Genetic Vectors / administration & dosage
Genetic Vectors / genetics
Genetic Vectors / therapeutic use
Heme Oxygenase-1 / genetics*
Micelles
Plasmids / administration & dosage*
Plasmids / genetics
Plasmids / therapeutic use
Rabbits
Rats
Rats, Sprague-Dawley
Regeneration*
Temperature
Transfection

Substances

Acrylic Resins
Micelles
poly-N-isopropylacrylamide
DNA
Heme Oxygenase-1