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Is There Additive Therapeutic Effect When GCSF Combined with Adipose-Derived Stem Cell in a Rat Model of Acute Spinal Cord Injury?

  • Min, Joongkee (Department of Neurological Surgery, Asan Medical Center, University of Ulsan College of Medicine) ;
  • Kim, Jeong Hoon (Department of Neurological Surgery, Asan Medical Center, University of Ulsan College of Medicine) ;
  • Choi, Kyoung Hyo (Department of Rehabilitation Medicine, Asan Medical Center, University of Ulsan College of Medicine) ;
  • Yoon, Hyung Ho (Department of Neurological Surgery, Asan Medical Center, University of Ulsan College of Medicine) ;
  • Jeon, Sang Ryong (Department of Neurological Surgery, Asan Medical Center, University of Ulsan College of Medicine)
  • Received : 2016.10.12
  • Accepted : 2016.12.09
  • Published : 2017.07.01

Abstract

Objective : Functional and neural tissue recovery has been reported in many animal studies conducted with stem cells. However, the combined effect of cytokines and stem cells has not yet been adequately researched. Here, we analyzed the additive effects of granulocyte colony-stimulating factor (GCSF) on adipose-derived stem cells (ADSCs) infusion in the treatment of acute spinal cord injury (SCI) in rats. Methods : Four days after intrathecal infusion tubes implantation in Sprague-Dawley rats, SCI was induced with an infinite horizon impactor. In the Sham group (n=5), phosphate-buffered saline was injected 3, 7, and 14 days after SCI. GCSF, ADSCs, and ADSCs with GCSF were injected at the same time in the GCSF (n=8), ADSC (n=8), and ADSC+GCSF groups (n=7), respectively. Results : The ADSC and ADSC+GCSF groups, but not the GCSF group, showed significantly higher Basso-Beattie-Bresnahan scores than the Sham group during 8 weeks (p<0.01), but no significant difference between the ADSC and ADSC+GCSF groups. In the ladder rung test, all four groups were significantly different from each other, with the ADSC+GCSF group showing the best improvement (p<0.01). On immunofluorescent staining (GAP43, MAP2), western blotting (GAP43), and reverse transcription polymerase chain reaction (GAP43, nerve growth factor), the ADSC and ADSC+GCSF groups showed higher levels than the Sham and GCSF groups. Conclusion : Our analyses suggest that the combination of GCSF and ADSCs infusions in acute SCI in the rat does not have a significant additive effect. Hence, when combination agents for SCI stem cell therapy are considered, molecules other than GCSF, or modifications to the methodology, should be investigated.

Keywords

References

  1. Aguilar RM, Steward O : A bilateral cervical contusion injury model in mice: assessment of gripping strength as a measure of forelimb motor function. Exp Neurol 221 : 38-53, 2010 https://doi.org/10.1016/j.expneurol.2009.09.028
  2. Barry FP : Biology and clinical applications of mesenchymal stem cells. Birth Defects Res C Embryo Today 69 : 250-256, 2003 https://doi.org/10.1002/bdrc.10021
  3. Basso DM, Beattie MS, Bresnahan JC : A sensitive and reliable locomotor rating scale for open field testing in rats. J Neurotrauma 12 : 1-21, 1995 https://doi.org/10.1089/neu.1995.12.1
  4. Beveridge RA, Miller JA, Kales AN, Binder RA, Robert NJ, Harvey JH, et al. : A comparison of efficacy of sargramostim (yeast-derived RhuGMCSF) and filgrastim (bacteria-derived RhuG-CSF) in the therapeutic setting of chemotherapy-induced myelosuppression. Cancer Invest 16 : 366-373, 1998 https://doi.org/10.3109/07357909809115775
  5. Choi JS, Leem JW, Lee KH, Kim SS, Suh-Kim H, Jung SJ, et al. : Effects of human mesenchymal stem cell transplantation combined with polymer on functional recovery following spinal cord hemisection in rats. Korean J Physiol Pharmacol 16 : 405-411, 2012 https://doi.org/10.4196/kjpp.2012.16.6.405
  6. Cizkova D, Rosocha J, Vanicky I, Jergova S, Cizek M : Transplants of human mesenchymal stem cells improve functional recovery after spinal cord injury in the rat. Cell Mol Neurobiol 26 : 1167-1180, 2006
  7. Corre J, Barreau C, Cousin B, Chavoin JP, Caton D, Fournial G, et al. : Human subcutaneous adipose cells support complete differentiation but not self-renewal of hematopoietic progenitors. J Cell Physiol 208 : 282-288, 2006 https://doi.org/10.1002/jcp.20655
  8. Couto PA, Filipe VM, Magalhaes LG, Pereira JE, Costa LM, Melo-Pinto P, et al. : A comparison of two-dimensional and three-dimensional techniques for the determination of hindlimb kinematics during treadmill locomotion in rats following spinal cord injury. J Neurosci Methods 173 : 193-200, 2008 https://doi.org/10.1016/j.jneumeth.2008.06.006
  9. Demetri GD, Griffin JD : Granulocyte colony-stimulating factor and its receptor. Blood 78 : 2791-2808, 1991
  10. Deng YB, Yuan QT, Liu XG, Liu XL, Liu Y, Liu ZG, et al. : Functional recovery after rhesus monkey spinal cord injury by transplantation of bone marrow mesenchymal-stem cell-derived neurons. Chin Med J (Engl) 118 : 1533-1541, 2005
  11. Dittgen T, Pitzer C, Plaas C, Kirsch F, Vogt G, Laage R, et al. : Granulocyte- colony stimulating factor (G-CSF) improves motor recovery in the rat impactor model for spinal cord injury. PLoS One 7 : e29880, 2012 https://doi.org/10.1371/journal.pone.0029880
  12. Furuya T, Hashimoto M, Koda M, Okawa A, Murata A, Takahashi K, et al. : Treatment of rat spinal cord injury with a Rho-kinase inhibitor and bone marrow stromal cell transplantation. Brain Res 1295 : 192-202, 2009 https://doi.org/10.1016/j.brainres.2009.07.087
  13. Gimble JM, Guilak F : Differentiation potential of adipose derived adult stem (ADAS) cells. Curr Top Dev Biol 58 : 137-160, 2003
  14. Hodgetts SI, Simmons PJ, Plant GW : A comparison of the behavioral and anatomical outcomes in sub-acute and chronic spinal cord injury models following treatment with human mesenchymal precursor cell transplantation and recombinant decorin. Exp Neurol 248 : 343-359, 2013 https://doi.org/10.1016/j.expneurol.2013.06.018
  15. Jeon SR, Park JH, Lee JH, Kim DY, Kim HS, Sung IY, et al. : Treatment of spinal cord injury with bone marrow-derived, cultured autologous mesenchymal stem cells. Tissue Eng Regen Med 7 : 316-322, 2010
  16. Jeong JH, Lee JH, Jin ES, Min JK, Jeon SR, Choi KH : Regeneration of intervertebral discs in a rat disc degeneration model by implanted adipose-tissue-derived stromal cells. Acta Neurochir (Wien) 152 : 1771-1777, 2010 https://doi.org/10.1007/s00701-010-0698-2
  17. Katz AJ, Tholpady A, Tholpady SS, Shang H, Ogle RC : Cell surface and transcriptional characterization of human adipose-derived adherent stromal (hADAS) cells. Stem Cells 23 : 412-423, 2005 https://doi.org/10.1634/stemcells.2004-0021
  18. Kawabe J, Koda M, Hashimoto M, Fujiyoshi T, Furuya T, Endo T, et al. : Neuroprotective effects of granulocyte colony-stimulating factor and relationship to promotion of angiogenesis after spinal cord injury in rats: laboratory investigation. J Neurosurg Spine 15 : 414-421, 2011 https://doi.org/10.3171/2011.5.SPINE10421
  19. Kim KN, Oh SH, Lee KH, Yoon DH : Effect of human mesenchymal stem cell transplantation combined with growth factor infusion in the repair of injured spinal cord. Acta Neurochir Suppl 99 : 133-136, 2006
  20. Kishk NA, Gabr H, Hamdy S, Afifi L, Abokresha N, Mahmoud H, et al. : Case control series of intrathecal autologous bone marrow mesenchymal stem cell therapy for chronic spinal cord injury. Neurorehabil Neural Repair 24 : 702-708, 2010 https://doi.org/10.1177/1545968310369801
  21. Lee HJ, Kim KS, Park IH, Kim SU : Human neural stem cells over-expressing VEGF provide neuroprotection, angiogenesis and functional recovery in mouse stroke model. PLoS One 2 : e156, 2007 https://doi.org/10.1371/journal.pone.0000156
  22. Lee KH, Suh-Kim H, Choi JS, Jeun SS, Kim EJ, Kim SS, et al. : Human mesenchymal stem cell transplantation promotes functional recovery following acute spinal cord injury in rats. Acta Neurobiol Exp (Wars) 67 : 13-22, 2007
  23. Liang P, Jin LH, Liang T, Liu EZ, Zhao SG : Human neural stem cells promote corticospinal axons regeneration and synapse reformation in injured spinal cord of rats. Chin Med J (Engl) 119 : 1331-1338, 2006
  24. Lim JH, Byeon YE, Ryu HH, Jeong YH, Lee YW, Kim WH, et al. : Transplantation of canine umbilical cord blood-derived mesenchymal stem cells in experimentally induced spinal cord injured dogs. J Vet Sci 8 : 275-282, 2007 https://doi.org/10.4142/jvs.2007.8.3.275
  25. Mukaetova-Ladinska EB, Andras A, Milne J, Abdel-All Z, Borr I, Jaros E, et al. : Synaptic proteins and choline acetyltransferase loss in visual cortex in dementia with Lewy bodies. J Neuropathol Exp Neurol 72 : 53-60, 2013 https://doi.org/10.1097/NEN.0b013e31827c5710
  26. Murphy JM, Fink DJ, Hunziker EB, Barry FP : Stem cell therapy in a caprine model of osteoarthritis. Arthritis Rheum 48 : 3464-3474, 2003 https://doi.org/10.1002/art.11365
  27. Nandoe Tewarie RD, Hurtado A, Ritfeld GJ, Rahiem ST, Wendell DF, Barroso MM, et al. : Bone marrow stromal cells elicit tissue sparing after acute but not delayed transplantation into the contused adult rat thoracic spinal cord. J Neurotrauma 26 : 2313-2322, 2009 https://doi.org/10.1089/neu.2009.0987
  28. Nishi RA, Liu H, Chu Y, Hamamura M, Su MY, Nalcioglu O, et al. : Behavioral, histological, and ex vivo magnetic resonance imaging assessment of graded contusion spinal cord injury in mice. J Neurotrauma 24 : 674-689, 2007 https://doi.org/10.1089/neu.2006.0204
  29. Pal R, Venkataramana NK, Bansal A, Balaraju S, Jan M, Chandra R, et al. : Ex vivo-expanded autologous bone marrow-derived mesenchymal stromal cells in human spinal cord injury/paraplegia: a pilot clinical study. Cytotherapy 11 : 897-911, 2009 https://doi.org/10.3109/14653240903253857
  30. Park JH, Kim DY, Sung IY, Choi GH, Jeon MH, Kim KK, et al. : Longterm results of spinal cord injury therapy using mesenchymal stem cells derived from bone marrow in humans. Neurosurgery 70 : 1238-1247; discussion 1247, 2012 https://doi.org/10.1227/NEU.0b013e31824387f9
  31. Parr AM, Kulbatski I, Tator CH : Transplantation of adult rat spinal cord stem/progenitor cells for spinal cord injury. J Neurotrauma 24 : 835-845, 2007 https://doi.org/10.1089/neu.2006.3771
  32. Pitzer C, Klussmann S, Kruger C, Letellier E, Plaas C, Dittgen T, et al. : The hematopoietic factor granulocyte-colony stimulating factor improves outcome in experimental spinal cord injury. J Neurochem 113 : 930-942, 2010 https://doi.org/10.1111/j.1471-4159.2010.06659.x
  33. Quertainmont R, Cantinieaux D, Botman O, Sid S, Schoenen J, Franzen R : Mesenchymal stem cell graft improves recovery after spinal cord injury in adult rats through neurotrophic and pro-angiogenic actions. PLoS One 7 : e39500, 2012 https://doi.org/10.1371/journal.pone.0039500
  34. Ritfeld GJ, Nandoe Tewarie RD, Vajn K, Rahiem ST, Hurtado A, Wendell DF, et al. : Bone marrow stromal cell-mediated tissue sparing enhances functional repair after spinal cord contusion in adult rats. Cell Transplant 21 : 1561-1575, 2012 https://doi.org/10.3727/096368912X640484
  35. Rodbell M : Metabolism of isolated fat cells. I. Effects of hormones on glucose metabolism and lipolysis. J Biol Chem 239 : 375-380, 1964
  36. Roussos I, Rodriguez M, Villan D, Ariza A, Rodriguez L, Garcia J : Development of a rat model of spinal cord injury and cellular transplantation. Transplant Proc 37 : 4127-4130, 2005 https://doi.org/10.1016/j.transproceed.2005.09.185
  37. Ruan H, Zarnowski MJ, Cushman SW, Lodish HF : Standard isolation of primary adipose cells from mouse epididymal fat pads induces inflammatory mediators and down-regulates adipocyte genes. J Biol Chem 278 : 47585-47593, 2003 https://doi.org/10.1074/jbc.M305257200
  38. Schaffler A, Buchler C : Concise review: adipose tissue-derived stromal cells--basic and clinical implications for novel cell-based therapies. Stem Cells 25 : 818-827, 2007 https://doi.org/10.1634/stemcells.2006-0589
  39. Scheff SW, Rabchevsky AG, Fugaccia I, Main JA, Lumpp JE Jr : Experimental modeling of spinal cord injury: characterization of a forcedefined injury device. J Neurotrauma 20 : 179-193, 2003 https://doi.org/10.1089/08977150360547099
  40. Shin DA, Kim JM, Kim HI, Yi S, Ha Y, Yoon DH, et al. : Comparison of functional and histological outcomes after intralesional, intracisternal, and intravenous transplantation of human bone marrow-derived mesenchymal stromal cells in a rat model of spinal cord injury. Acta Neurochir (Wien) 155 : 1943-1950, 2013 https://doi.org/10.1007/s00701-013-1799-5
  41. Shyu WC, Lin SZ, Yang HI, Tzeng YS, Pang CY, Yen PS, et al. : Functional recovery of stroke rats induced by granulocyte colony-stimulating factorstimulated stem cells. Circulation 110 : 1847-1854, 2004 https://doi.org/10.1161/01.CIR.0000142616.07367.66
  42. Sofroniew MV, Howe CL, Mobley WC : Nerve growth factor signaling, neuroprotection, and neural repair. Annu Rev Neurosci 24 : 1217-1281, 2001 https://doi.org/10.1146/annurev.neuro.24.1.1217
  43. Suh HI, Min J, Choi KH, Kim SW, Kim KS, Jeon SR : Axonal regeneration effects of Wnt3a-secreting fibroblast transplantation in spinal cordinjured rats. Acta Neurochir (Wien) 153 : 1003-1010, 2011 https://doi.org/10.1007/s00701-011-0945-1
  44. Sykova E, Jendelova P, Urdzikova L, Lesny P, Hejcl A : Bone marrow stem cells and polymer hydrogels-two strategies for spinal cord injury repair. Cell Mol Neurobiol 26 : 1113-1129, 2006
  45. Thuret S, Moon LD, Gage FH : Therapeutic interventions after spinal cord injury. Nat Rev Neurosci 7 : 628-643, 2006 https://doi.org/10.1038/nrn1955
  46. Tobias CA, Han SS, Shumsky JS, Kim D, Tumolo M, Dhoot NO, et al. : Alginate encapsulated BDNF-producing fibroblast grafts permit recovery of function after spinal cord injury in the absence of immune suppression. J Neurotrauma 22 : 138-156, 2005 https://doi.org/10.1089/neu.2005.22.138
  47. Urdzikova L, Jendelova P, Glogarova K, Burian M, Hajek M, Sykova E : Transplantation of bone marrow stem cells as well as mobilization by granulocyte-colony stimulating factor promotes recovery after spinal cord injury in rats. J Neurotrauma 23 : 1379-1391, 2006 https://doi.org/10.1089/neu.2006.23.1379
  48. Weaver CH, Buckner CD, Longin K, Appelbaum FR, Rowley S, Lilleby K, et al. : Syngeneic transplantation with peripheral blood mononuclear cells collected after the administration of recombinant human granulocyte colony-stimulating factor. Blood 82 : 1981-1984, 1993
  49. Wright KT, El Masri W, Osman A, Chowdhury J, Johnson WE : Concise review: Bone marrow for the treatment of spinal cord injury: mechanisms and clinical applications. Stem Cells 29 : 169-178, 2011 https://doi.org/10.1002/stem.570
  50. Zhou Z, Chen Y, Zhang H, Min S, Yu B, He B, et al. : Comparison of mesenchymal stromal cells from human bone marrow and adipose tissue for the treatment of spinal cord injury. Cytotherapy 15 : 434-448, 2013 https://doi.org/10.1016/j.jcyt.2012.11.015

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