Release of Plasmid DNA-Encoding IL-10 from PLGA Microparticles Facilitates Long-Term Reversal of Neuropathic Pain Following a Single Intrathecal Administration
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Interleukin-10 (IL-10) is an anti-inflammatory molecule that has achieved interest as a therapeutic for neuropathic pain. In this work, the potential of plasmid DNA-encoding IL-10 (pDNA-IL-10) slowly released from biodegradable microparticles to provide long-term pain relief in an animal model of neuropathic pain was investigated.
PLGA microparticles encapsulating pDNA-IL-10 were developed and assessed both in vitro and in vivo.
In vitro, pDNA containing microparticles activated macrophages, enhanced the production of nitric oxide, and increased the production of IL-10 protein relative to levels achieved with unencapsulated pDNA-IL-10. In vivo, intrathecally administered microparticles embedded in meningeal tissue, induced phagocytic cell recruitment to the cerebrospinal fluid, and relieved neuropathic pain for greater than 74 days following a single intrathecal administration, a feat not achieved with unencapsulated pDNA. Therapeutic effects of microparticle-delivered pDNA-IL-10 were blocked in the presence of IL-10-neutralizing antibody, and elevated levels of plasmid-derived IL-10 were detected in tissues for a prolonged time period post-injection (>28 days), demonstrating that therapeutic effects are dependent on IL-10 protein production.
These studies demonstrate that microparticle encapsulation significantly enhances the potency of intrathecally administered pDNA, which may be extended to treat other disorders that require intrathecal gene therapy.
Key wordsinterleukin-10 microparticle pDNA PLGA neuropathic pain
anti-rat interleukin-10 IgG neutralizing antibody
chronic constriction injury
limulus amebocyte lysate
major histocompatibility complex-II
Funding for this work was provided by NIH grant DA018156, and E. Dengler was supported by NSF Grant DGE-0549500. The authors would like to acknowledge the expert assistance of Dr. Travis Hughes and Dr. Leslie Leinwand at the University of Colorado for consultations on plasmid DNA growth and characterization. We would also like to thank Avigen Inc. (Alameda, CA) for the purification of Anti-IL-10-IgG and Control-IgG antibodies used in this work and Jenny L. Wilkerson at the University of New Mexico for her assistance with surgical procedures.
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