EFFETS OF CHONDROITINASE ABC COMBINED WITH BONE MARROW MESENCHYMAL STEM CELLS TRANSPLANTATION ON REPAIR OF SPINAL CORD INJURY IN RATS_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

ZHANG Chun , HE Xijing , LI Haopeng.

Department of Orthopedics II, the Second Hospital Affiliated to the Medical College of Xi’an Jiaotong University, Xi’an Shaanxi, 710004, P.R.China. Corresponding author: HE Xijing, E-mail: xijing_h@vip.tom.com;

Corresponding author：

Keywords：

Bone marrow mesenchymal stem cells; Chondroitinase ABC; Spinal cord injury; Cell therapy; Rat

DOI：

10.7507/1002-1892.20130121

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Objective To investigate the effects of chondroitinase ABC (ChABC) combined with bone marrow mesenchymal stem cells (BMSCs) in repair spinal cord injury of rats. Methods Primary BMSCs were isolated and cultured from the femur and tibia of neonatal Sprague Dawley (SD) rats. The spinal cord injury model was established in 24 adult SD male rats (weighing, 200-230 g), which were randomly divided into control group (group A), BMSCs transplantation group (group B), ChABC injection group (group C), and ChABC and BMSCs transplantation group (group D), 6 rats in each group. At 7 and 14 days after injury, Basso-Beattie-Bresnahan (BBB) score criteria was used to evaluate the hindlimb motor function; at 14 days after injury, the injured spinal cord tissue was perfused and stained by HE for further calculation of the injury area. Immunofluorescence staining were used for observing the expressions of glial fibrillary acidic protein (GFAP)/chondroitin sulfate proteoglycan (CSPG) and GFAP/growth associated protein 43 (GAP43). Results At 7 days after injury, three joints movement of the hindlimbs were recovered in all groups, and no significant difference in the BBB score was found among 4 groups (P gt; 0.05). At 14 days after injury, no load drag was observed in 3 joints of the hindlimbs in groups A, B, and C, but weight-bearing plantar or occasional dorsalis pedis weight-bearing walking was observed in group D with no plantar walking. The BBB score of group D was significantly higher than that of the other 3 groups (P lt; 0.05). HE staining showed that the cavity formed in the damage zone, and there were a large number of macrophages in the cavity and its surrounding, which was wrapped by scar tissue. The damage area of group D was significantly smaller than that of the other 3 groups (P lt; 0.05). At 14 days after injury, the GFAP/CSPG double immunofluorescence staining showed that the astroglial scar damage zone in group D was significantly reduced, and no cavity formation was found. And the fluorescence intensity in groups C and D was significantly lower than that in group B (P lt; 0.05). The GFAP/GAP43 double immunofluorescence staining showed that GAP43-positive fibers passed through the damage zone in group D and the fluorescence intensity in group D was significantly higher than those in groups B and C (P lt; 0.05). Conclusion Inhibition of astrocytes secreting CSPG by ChABC combined with BMSCs transplantation in early injury may promote the regeneration of nerve fibers, and repair spinal cord injury in rats.

Citation： ZHANG Chun,HE Xijing,LI Haopeng.. EFFETS OF CHONDROITINASE ABC COMBINED WITH BONE MARROW MESENCHYMAL STEM CELLS TRANSPLANTATION ON REPAIR OF SPINAL CORD INJURY IN RATS. Chinese Journal of Reparative and Reconstructive Surgery, 2013, 27(5): 541-546. doi: 10.7507/1002-1892.20130121 Copy

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12.	祖波, 尹宗生. 机械性脊髓损伤模型的建立及其评价. 中国临床康复, 2006, 10(12): 151-153.
13.	Coumans JV, Lin TT, Dai HN, et al. Axonal regeneration and functional recovery after complete spinal cord transection in rats by delayed treatment with transplants and neurotrophins. J Neurosci, 2001, 21(23): 9334-9344.
14.	Benowitz LI, Popovich PG. Inflammation and axon regeneration. Curr Opin Neurol, 2011, 24(6): 577-583.
15.	游思维, 苏国辉. 促进中枢神经损伤再生的探索之路. 中国处方药, 2004, 3(2): 30-33.
16.	Rhodes KE, Moon LD, Fawcett JW. Inhibiting cell proliferation during formation of the glial scar effect on axon regeneration in the CNS. Neuroscience, 2003, 120(1): 41-56.
17.	Tang X, Davies JE, Davies SJ. Changes in distribution, cell associations and protein expression levels of NG2, neurocan, phosphacan, brevican, versican V2 and tenascin-C during acute to chronic maturation of spinal cord scar. J Neurosci Res, 2003, 71(3): 427-444.
18.	Minor K, Tang X, Kahrilas G, et al. Decorin promotes robust axon growth on inhibitory CSPGs and myelin via a direct effect on neurons. Neuroboil Dis, 2008, 32(1): 88-95.
19.	Willerth SM, Sakiyama-Elbert SE. Cell therapyfor spinal cord regeneration. Adv Drug Deliv Rev, 2008, 60(2): 263-276.
20.	Evans GR. Peripheral nerve injury: a review and approach to tissue engineered constructs. Anay Rec, 2001, 263(4): 396-404.
21.	Ruff CA, Wilcox JT, Fehlings MG, et al. Cell-based transplantation strategies to promote plasticity following spinal cord injury. Exp Neurol, 2012, 235(1): 78-90.
22.	Wu B, Sun L, Li P, et al. Transplantation of oligodendrocyte precursor cells improves myelination and promotes functional recovery after spinal cord injury. Injury, 2012, 43(6): 794-801.

1. 沈学峰. 大鼠脊髓损伤后二次继发性损伤的发现及其治疗. 西安: 第四军医大学: 2006.
2. 焦西英, 程希平, 钱新宏, 等. 脊髓坏死面积的测量方法介绍. 神经解剖学杂志, 2006, 22(3): 356-358.
3. Fawcett JW, Asher RA. The glial scar and central nervous system repair. Brain Res Bull, 1999, 49(6): 377-391.
4. Hofstetter CP, Schwarz EJ, Hess D, et al. Marrow stromal cells from guiding strands in the injuryed spinal cord and promote recovery. Proc Natl Acad Sci U S A, 2002, 99(4): 2199-2204.
5. Woodlbury D, Schwarz EJ, Proclop DJ, et al. Adult rat and hunan bone marrow stromal cells ditterentiate into neurons. J Neurosci Res, 2000, 61(4): 364-370.
6. Novikova LN, Brohlin M, Kingham PJ, et al. Neuroprotective and growth-promoting effects of bone marrow stromal cells after cervical spinal cord injury in adult rats. Cytotherapy, 2011, 13(7): 873-887.
7. Zurita M, Otero L, Aguayo C, et al. Cell therapy for spinal cord repair: optimization of biologic scaffolds for survival and neural differentiation of human bone marrow stromal cells. Cytotherapy, 2010, 12(4): 522-537.
8. Park KW, Eglitis MA, Mouradian MM. Protection of migral neurous by GDNF-engineered marrow cell trancplantation. Neurosci Res, 2001, 40(4): 315-323.
9. Yazdani SD, Pedram M, Hafizi M, et al. A comparison between neurally induced bone marrow derived mesenchymal stem cells and olfactory ensheathing glial cells to repair spinal cord injuries in rats. Tissue Cell, 2012, 44(4): 205-213.
10. Pal R, Gopinath C, Rao NM, et al. Functional recovery after transplantation of bone marrow-derived human mesenchymal stromal cells in a rat model of spinal cord injury. Cytotherapy, 2010, 12(6): 792-806.
11. Zhang JF, Zhao FS, Wu G, et al. Therapeutic effect of co-transplantation of neuregulin 1-transfected-Schwann cells and bone marrow stromal cells on spinal cord hemisection syndrome. Neuroscience, 2011, 497(2): 128-133.
12. 祖波, 尹宗生. 机械性脊髓损伤模型的建立及其评价. 中国临床康复, 2006, 10(12): 151-153.
13. Coumans JV, Lin TT, Dai HN, et al. Axonal regeneration and functional recovery after complete spinal cord transection in rats by delayed treatment with transplants and neurotrophins. J Neurosci, 2001, 21(23): 9334-9344.
14. Benowitz LI, Popovich PG. Inflammation and axon regeneration. Curr Opin Neurol, 2011, 24(6): 577-583.
15. 游思维, 苏国辉. 促进中枢神经损伤再生的探索之路. 中国处方药, 2004, 3(2): 30-33.
16. Rhodes KE, Moon LD, Fawcett JW. Inhibiting cell proliferation during formation of the glial scar effect on axon regeneration in the CNS. Neuroscience, 2003, 120(1): 41-56.
17. Tang X, Davies JE, Davies SJ. Changes in distribution, cell associations and protein expression levels of NG2, neurocan, phosphacan, brevican, versican V2 and tenascin-C during acute to chronic maturation of spinal cord scar. J Neurosci Res, 2003, 71(3): 427-444.
18. Minor K, Tang X, Kahrilas G, et al. Decorin promotes robust axon growth on inhibitory CSPGs and myelin via a direct effect on neurons. Neuroboil Dis, 2008, 32(1): 88-95.
19. Willerth SM, Sakiyama-Elbert SE. Cell therapyfor spinal cord regeneration. Adv Drug Deliv Rev, 2008, 60(2): 263-276.
20. Evans GR. Peripheral nerve injury: a review and approach to tissue engineered constructs. Anay Rec, 2001, 263(4): 396-404.
21. Ruff CA, Wilcox JT, Fehlings MG, et al. Cell-based transplantation strategies to promote plasticity following spinal cord injury. Exp Neurol, 2012, 235(1): 78-90.
22. Wu B, Sun L, Li P, et al. Transplantation of oligodendrocyte precursor cells improves myelination and promotes functional recovery after spinal cord injury. Injury, 2012, 43(6): 794-801.

Chinese Journal of Reparative and Reconstructive Surgery

EFFETS OF CHONDROITINASE ABC COMBINED WITH BONE MARROW MESENCHYMAL STEM CELLS TRANSPLANTATION ON REPAIR OF SPINAL CORD INJURY IN RATS

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