Locally Applied Stem Cell Exosome-Scaffold Attenuates Nerve Injury-Induced Pain in Rats

Abstract

Jong-Ming Hsu,1– 3,* Sheng-Jie Shiue,4,* Kuender D Yang,5– 7,* Han-Shiang Shiue,8 Yi-Wei Hung,4 Pavani Pannuru,4 Raju Poongodi,4 Hsin-Yi Lin,9,10 Jen-Kun Cheng3,4,11 1Department of Urology, Mackay Memorial Hospital, Taipei 10449, Taiwan; 2Mackay Junior College of Medicine, Nursing, and Management, Taipei 11260, Taiwan; 3Department of Medicine, Mackay Medical College, New Taipei City 252, Taiwan; 4Department of Medical Research, Mackay Memorial Hospital, Taipei 10449, Taiwan; 5Institute of Biomedical Science, Mackay Medical College, New Taipei City 252, Taiwan; 6Department of Pediatrics, Mackay Memorial Hospital, Taipei 10449, Taiwan; 7Institute of Clinical Medicine, National Yang-Ming University, Taipei 11221, Taiwan; 8Institute of Neuroscience, National Yang-Ming University, Taipei 11221, Taiwan; 9Department of Chemical Engineering and Biotechnology, National Taipei University of Technology, Taipei 106, Taiwan; 10Graduate Institute of Biochemical and Biomedical Engineering, National Taipei University of Technology, Taipei 106, Taiwan; 11Department of Anesthesiology, Mackay Memorial Hospital, Taipei 10449, Taiwan*These authors contributed equally to this workCorrespondence: Hsin-Yi LinDepartment of Chemical Engineering and Biotechnology, National Taipei University of Technology, No. 1, Sec 3, Zhongxiao E. Road, Taipei 106, Taiwan, Email [email protected] ChengDepartment of Anesthesiology, Mackay Memorial Hospital, No. 92, Sec. 2, Zhongshan N. Road, Taipei City 10449, TaiwanEmail [email protected]: Nerve injury-induced pain is difficult to treat. In this study, we developed an alginate scaffold with human umbilical cord mesenchymal stem cell exosomes (EX-SC) to treat nerve injury-induced pain.Materials and Methods: The scaffold was prepared and characterized for its physical traits and biocompatibility. In vitro studies of PC12 and HEK293 cells were used to evaluate the neuroprotective and neurotrophic effects of exosomes. Right L5/6 spinal nerve ligation (SNL) was performed in Sprague-Dawley rats to induce mechanical allodynia and thermal hyperalgesia, evaluated by von Frey hair and radiant heat tests. The EX-SC was wrapped around ligated L5/6 spinal nerves for treatment. Western blotting and immunofluorescence staining were used to evaluate neuron/glial activation, cytokines and neurotrophic factor of affected dorsal root ganglion (DRG).Results: In cell culture assay, the exosomes induce neurite outgrowth of PC12 cells and protect PC12 and HEK293 cells against formaldehyde acid treatment. On post-ligation day 21, rats receiving EX-SC had significantly higher median (interquartile range) withdrawal threshold and latency [14.1 (13.7– 15.5) g, 14.2 (13.7– 15.3) s] than saline-SC-treated rats [2.1 (1.7– 3.0) g, 2.0 (1.8– 2.4) s, P=0.02 and 0.002]. The EX-SC also attenuated SNL-induced up-regulation of c-Fos, GFAP, Iba1, TNF-α and IL-1β, while enhancing the level of IL-10 and GDNF, in the ipsilateral L5/6 DRG. After implantation for 21 days, the EX-SC enhanced the expression of myelin basic protein and IL-10 in injured L5/6 axons.Conclusion: We demonstrate the EX-SC possesses antinociceptive, anti-inflammation and pro-neurotrophic effects in the SNL pain model. It could be a promising therapeutic alternative for nerve injury-induced pain.Keywords: exosome, nerve injury, neuropathic pain, scaffold, spinal nerve ligation, stem cell

Keywords

• exosome
• nerve injury
• neuropathic pain
• scaffold
• spinal nerve ligation
• stem cell.