Neurotrophin-3 modified hyaluronan-methylcellulose hydrogel promotes neurological function in rats with spinal cord injury

doi:10.3969/j.issn.2095-4344.1640

Abstract

Abstract:

BACKGROUND: Hyaluronan-methylcellulose hydrogel cannot only be conjugated with short peptide sequences and growth factors to achieve sustained release, but also has a role in blocking dural defects and reducing inflammation. It is an ideal biomaterial for the treatment of spinal cord injury.

OBJECTIVE: To investigate the effect of neurotrophin-3 modified hyaluronan-methylcellulose (HAMC-NT-3) hydrogel on the recovery of neurological function in rats with spinal cord injury.

METHODS: Fifty-four female Sprague-Dawley rats (provided by the Experimental Animal Center of the Academy of Military Medical Sciences in China) were randomly divided into three groups (n=18 per group). The sham group only underwent T₁₀ laminectomy. In the model group and the experimental group, an aneurysm clip was used to establish spinal cord injury models after T₁₀ laminectomy. The experimental group was locally injected with HAMC-NT-3 hydrogel. The Basso Beattie Bresnahan function scoring was performed at 1 day, 1, 2, 3, 4, 5, 6, 7, and 8 weeks after surgery. The inclined plane test was performed at 4, 6 and 8 weeks after surgery to evaluate the recovery of hindlimb motor function. ELISA was used to detect the concentrations of inflammatory factors in the spinal cord at 1 week after surgery. Immunohistochemical staining was used to observe the area of syringomyelia, glial fibrillary acidic protein expression and nerve regeneration at 8 weeks after surgery.

RESULTS AND CONCLUSION: (1) The Basso Beattie Bresnahan scores of the model group and the experimental group were lower than those of the sham group at various time points after surgery (P < 0.05). The Basso Beattie Bresnahan scores of the experimental group were higher than those of the model group at 4-8 weeks after surgery (P < 0.05). (2) In the inclined plane test, the maximum inclined angles of the model group and the experimental group at each time point after surgery were lower than that of the sham group (P < 0.05). The maximum inclined angles of the experimental group at 6 and 8 weeks after surgery were higher than those of the sham group (P < 0.05). (3) The concentrations of tumor necrosis factor-α, interleukin-1β, interleukin-6 and interleukin-10 in the experimental group and the model group were higher than those in the sham group (P < 0.05). The concentrations of tumor necrosis factor-α, interleukin-1β and interleukin-6 in the experimental group were lower than those in the model group (P < 0.05). The concentration of interleukin-10 in the experimental group was higher than that in the model group (P < 0.05). (4) Immunohistochemical staining showed that the expression levels of glial fibrillary acidic protein in the experimental group and the model group were higher than those in the sham group, while the expression of glial fibrillary acidic protein in the experimental group was lower than that in the model group. The area of syringomyelia in the experimental group was smaller than that in the model group (P < 0.05). These results indicate that local injection of HAMC-NT-3 hydrogel can effectively inhibit inflammation as well as astrocyte activation and proliferation, reduce fibrous scar formation, and promote the protection of nerve tissue and the recovery of hindlimb motor function after spinal cord injury.

Key words: Hydrogel, Hyaluronic Acid, Neurotrophin 3, Methylcellulose, Spinal Cord Injuries, Tissue Engineering

CLC Number:

R459.9

R318.08

He Zhijiang, Zhu Lei, Cheng Shixiang, Huang Kui, Chen Cao, Sun Minglin. Neurotrophin-3 modified hyaluronan-methylcellulose hydrogel promotes neurological function in rats with spinal cord injury[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(14): 2202-2207.

Figures/Tables 4

References

[1] Ackery A,Tator C,Krassioukov A.A global perspective on spinal cord injury epidemiology.J Neurotrauma. 2004;21(10):1355-1370.

[2] Mothe AJ,Tam RY,Zahir T,et al.Repair of the injured spinal cord by transplantation of neural stem cells in a hyaluronan-based hydrogel. Biomaterials. 2013;34(15):3775-3783.

[3] Moon YJ,Lee JY,Oh MS,et al.Inhibition of inflammation and oxidative stress by Angelica dahuricae radix extract decreases apoptotic cell death and improves functional recovery after spinal cord injury. J Neurosci Res. 2012;90(1):243-256.

[4] An Y,Tsang KK,Zhang H.Potential of stem cell based therapy and tissue engineering in the regeneration of the central nervous system.Biomed Mater.2006;1(2):R38-344.

[5] Blesch A,Tuszynski MH.Spinal cord injury: plasticity, regeneration and the challenge of translational drug development.Trends Neurosci. 2009; 32(1):41-47.

[6] Cregg JM,DePaul MA,Filous AR,et al.Functional regeneration beyond the glial scar.Exp Neurol.2014;253:197-207.

[7] Li G,Che MT,Zeng X,et al.Neurotrophin-3 released from implant of tissue-engineered fibroin scaffolds inhibits inflammation, enhances nerve fiber regeneration, and improves motor function in canine spinal cord injury.J Biomed Mater Res A.2018;106(8):2158-2170.

[8] Shibayama M,Hattori S,Himes BT,et al.Neurotrophin-3 prevents death of axotomized Clarke's nucleus neurons in adult rat.J Comp Neurol. 1998; 390(1):102-111.

[9] McMahon SS,Nikolskaya N,Choileain SN,et al.Thermosensitive hydrogel for prolonged delivery of lentiviral vector expressing neurotrophin-3 in vitro. J Gene Med.2011;13(11):591-601.

[10] Elliott Donaghue I,Tator CH,Shoichet MS.Sustained delivery of bioactive neurotrophin-3 to the injured spinal cord.Biomater Sci.2015;3(1):65-72.

[11] Li G,Che MT,Zhang K,et al.Graft of the NT-3 persistent delivery gelatin sponge scaffold promotes axon regeneration, attenuates inflammation, and induces cell migration in rat and canine with spinal cord injury. Biomaterials.2016;83:233-248.

[12] Ejstrup R,Kiilgaard JF,Tucker BA,et al.Pharmacokinetics of intravitreal glial cell line-derived neurotrophic factor: experimental studies in pigs. Exp Eye Res.2010;91(6):890-895.

[13] Baumann MD,Kang CE,Tator CH,et al.Intrathecal delivery of a polymeric nanocomposite hydrogel after spinal cord injury.Biomaterials. 2010; 31(30):7631-7639.

[14] Tam RY,Cooke MJ,Shoichet MS.A covalently modified hydrogel blend of hyaluronan–methyl cellulose with peptides and growth factors influences neural stem/progenitor cell fate.J Mater Chem.2012;22(37):19402.

[15] Basso DM, Beattie MS, Bresnahan JC. A sensitive and reliable locomotor rating scale for open field testing in rats.J Neurotrauma.1995;12(1):1-21.

[16] Liu S,Sandner B,Schackel T,et al.Regulated viral BDNF delivery in combination with Schwann cells promotes axonal regeneration through capillary alginate hydrogels after spinal cord injury.Acta Biomater. 2017; 60:167-180.

[17] Bartus K,James ND,Didangelos A,et al.Large-scale chondroitin sulfate proteoglycan digestion with chondroitinase gene therapy leads to reduced pathology and modulates macrophage phenotype following spinal cord contusion injury.J Neurosci.2014;34(14):4822-4836.

[18] Fouad K,Ghosh M,Vavrek R,et al.Dose and chemical modification considerations for continuous cyclic AMP analog delivery to the injured CNS.J Neurotrauma. 2009;26(5):733-740.

[19] Oh JS,An SS,Gwak SJ,et al.Hypoxia-specific VEGF-expressing neural stem cells in spinal cord injury model.Neuroreport.2012;23(3):174-178.

[20] Hong LTA,Kim YM,Park HH,et al.An injectable hydrogel enhances tissue repair after spinal cord injury by promoting extracellular matrix remodeling. Nat Commun.2017;8(1):533.

[21] 卫巍,刘燕飞,何洋,等.β折叠型自组装短肽水凝胶特性及在神经组织工程中的应用前景[J].中国组织工程研究, 2018,22(10):1586-1592.

[22] Cigognini D, Silva D, Paloppi S, et al.Evaluation of mechanical properties and therapeutic effect of injectable self-assembling hydrogels for spinal cord injury.J Biomed Nanotechnol.2014;10(2): 309-323.

[23] Liu Y, Ye H, Satkunendrarajah K, et al.A self-assembling peptide reduces glial scarring, attenuates post-traumatic inflammation and promotes neurological recovery following spinal cord injury.Acta Biomater. 2013; 9(9):8075-8088.

[24] Saghazadeh A,Rezaei N.The role of timing in the treatment of spinal cord injury. Biomed Pharmacother.2017;92:128-139.

[25] Coll-Miro M,Francos-Quijorna I,Santos-Nogueira E,et al.Beneficial effects of IL-37 after spinal cord injury in mice.Proc Natl Acad Sci U S A. 2016;113(5):1411-1416.

[26] Wakao N,Imagama S,Zhang H,et al.Hyaluronan oligosaccharides promote functional recovery after spinal cord injury in rats.Neurosci Lett. 2011;488(3):299-304.

[27] Wang CX, Olschowka JA, Wrathall JR. Increase of interleukin-1beta mRNA and protein in the spinal cord following experimental traumatic injury in the rat.Brain Res.1997;759(2):190-196.

[28] Lacroix S,Chang L,Rose-John S,et al.Delivery of hyper-interleukin-6 to the injured spinal cord increases neutrophil and macrophage infiltration and inhibits axonal growth. J Comp Neurol.2002;454(3):213-228.

[29] Ropper AE,Ropper AH.Acute Spinal Cord Compression.N Engl J Med. 2017;376(14):1358-1369.

[30] 秦茂林,杨卫兵,李红丽,等.小鼠胚胎干细胞NT3基因转染后移植对脊髓损伤的恢复作用[J].中国组织工程研究, 2008,12(12):2263-2266.

[31] 邓兴力,梁袁昕,杨智勇,等.神经营养因子3基因修饰神经干细胞移植脊髓损伤的相关蛋白表达[J].中国组织工程研究, 2010,14(36):6751-6754.

[32] Anderson MA,Burda JE,Ren Y,et al.Astrocyte scar formation aids central nervous system axon regeneration. Nature. 2016;532(7598):195-200.