A novel tissue-engineered crosslinked collagen-chitosan scaffold and its biocompatibility with nerve cells

doi:10.3969/j.issn.2095-4344.2017.18.020

Abstract

Abstract:

BACKGROUND: Treatment for peripheral nerve injury is still a challenge in the field of regenerative medicine. The use of scaffold materials is considered to be one of the promising opportunities to settle these defects. The ideal scaffold material should be safe and durable, but have no antigenicity.

OBJECTIVE: To develop a novel tissue-engineered collagen-chitosan composite scaffolds under modification in attempt to repair sciatic nerve injury in rats.

METHODS: First the collagen-chitosan composite scaffold was prepared, and then cross-linked with 1-ethyl-3-(3-2 methyl amino propyl)-WC 2 imide (EDC) and N-hydroxy succinimide (NHS). After modification, nerve cells were seeded onto the scaffold. Finally, the cell-scaffold composite was implanted into the injured sciatic nerve.

RESULTS AND CONCLUSION: The EDC/NHS-crosslinked collagen-chitosan scaffold showed a three-dimensional reticular structure. Four weeks after the cell-scaffold composite implantation, neurofilament protein 200 was found to express in the injured sciatic nerve. This indicates that the crosslinked collagen-chitosan scaffold has good cytocompatibility.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

Key words: Biocompatible Materials, Collagen, Chitosan, Tissue Engineering

CLC Number:

R318

Lu Jiang, Chen Xuan-li, Tang He-xin, She Na, Zhu Yao, Li Si-xing, Wang Tian-ying, Zhang Guo-hui.

A novel tissue-engineered crosslinked collagen-chitosan scaffold and its biocompatibility with nerve cells [J]. Chinese Journal of Tissue Engineering Research, 2017, 21(18): 2913-2919.

Figures/Tables 6

References

[1]Grinsell D, Keating CP. Peripheral nerve reconstruction after injury: a review of clinical and experimental therapies. Biomed Res Int. 2014;2014:698256.

[2]Panagopoulos GN, Megaloikonomos PD, Mavrogenis AF. The Present and Future for Peripheral Nerve Regeneration. Orthopedics. 2017;40(1):e141-e156.

[3]Faroni A, Mobasseri SA, Kingham PJ, et al. Peripheral nerve regeneration: experimental strategies and future perspectives. Adv Drug Deliv Rev. 2015;82-83:160-167.

[4]Allodi I, Udina E, Navarro X. Specificity of peripheral nerve regeneration: interactions at the axon level. Prog Neurobiol. 2012;98(1):16-37.

[5]Jiang L, Jones S, Jia X. Stem Cell Transplantation for Peripheral Nerve Regeneration: Current Options and Opportunities. Int J Mol Sci. 2017;18(1). pii: E94.

[6]Monaco G, Cholas R, Salvatore L, et al. Sterilization of collagen scaffolds designed for peripheral nerve regeneration: Effect on microstructure, degradation and cellular colonization. Mater Sci Eng C Mater Biol Appl. 2017;71:335-344.

[7]Kanetis L, Exarchou V, Charalambous Z, et al. Edible coating composed of chitosan and Salvia fruticosa Mill. extract for the control of grey mould of table grapes. J Sci Food Agric. 2017; 97(2):452-460.

[8]Gratzer PF, Santerre JP, Lee JM. The effect of chemical modification of amino acid side-chains on collagen degradation by enzymes. J Biomed Mater Res B Appl Biomater. 2007;81(1):1-11.

[9]Minardi S, Taraballi F, Wang X, et al. Biomimetic collagen/elastin meshes for ventral hernia repair in a rat model. Acta Biomater. 2017;50:165-177.

[10]Jithendra P, Rajam AM, Kalaivani T, et al. Preparation and characterization of aloe vera blended collagen-chitosan composite scaffold for tissue engineering applications. ACS Appl Mater Interfaces. 2013;5(15):7291-7298.

[11]Grover CN, Cameron RE, Best SM. Investigating the morphological, mechanical and degradation properties of scaffolds comprising collagen, gelatin and elastin for use in soft tissue engineering. J Mech Behav Biomed Mater. 2012;10:62-74.

[12]Davidenko N, Schuster CF, Bax DV, et al. Control of crosslinking for tailoring collagen-based scaffolds stability and mechanics. Acta Biomater. 2015;25:131-142.

[13]Kumari S, Kumar Annamareddy SH, Abanti S, et al. Physicochemical properties and characterization of chitosan synthesized from fish scales, crab and shrimp shells. Int J Biol Macromol. 2017. in press.

[14]Varun TK, Senani S, Jayapal N, et al. Extraction of chitosan and its oligomers from shrimp shell waste, their characterization and antimicrobial effect. Vet World. 2017;10(2):170-175.

[15]Marei NH, El-Mazny W, El-Shaer A, et al. Enhanced wound healing activity of desert locust (Schistocerca gregaria) vs. shrimp (Penaeus monodon) chitosan based scaffolds. Int J Biol Macromol. 2017;97:23-33.

[16]Zhang H, Yun S, Song L, et al. The preparation and characterization of chitin and chitosan under large-scale submerged fermentation level using shrimp by-products as substrate. Int J Biol Macromol. 2017;96:334-339.

[17]Wrobel S, Serra SC, Ribeiro-Samy S, et al. In vitro evaluation of cell-seeded chitosan films for peripheral nerve tissue engineering. Tissue Eng Part A. 2014;20(17-18):2339-2349.

[18]Han F, Dong Y, Su Z, et al. Preparation, characteristics and assessment of a novel gelatin-chitosan sponge scaffold as skin tissue engineering material. Int J Pharm. 2014;476(1-2): 124-133.

[19]Mututuvari TM, Harkins AL, Tran CD. Facile synthesis, characterization, and antimicrobial activity of cellulose-chitosan-hydroxyapatite composite material: a potential material for bone tissue engineering. J Biomed Mater Res A. 2013;101(11):3266-3277.

[20]Yin A, Zhang K, McClure MJ, et al. Electrospinning collagen/chitosan/poly(L-lactic acid-co-ε-caprolactone) to form a vascular graft: mechanical and biological characterization. J Biomed Mater Res A. 2013;101(5):1292-1301.

[21]Su JY, Chen SH, Chen YP, et al. Evaluation of Magnetic Nanoparticle-Labeled Chondrocytes Cultivated on a Type II Collagen-Chitosan/Poly(Lactic-co-Glycolic) Acid Biphasic Scaffold. Int J Mol Sci. 2017;18(1). pii: E87.

[22]Zhang ML, Cheng J, Xiao YC, et al. Raloxifene microsphere-embedded collagen/chitosan/β-tricalcium phosphate scaffold for effective bone tissue engineering. Int J Pharm. 2017;518(1-2):80-85.

[23]Muthukumar T, Aravinthan A, Sharmila J, et al. Collagen/chitosan porous bone tissue engineering composite scaffold incorporated with Ginseng compound K. Carbohydr Polym. 2016;152:566-574.

[24]Fan X, Chen K, He X, et al. Nano-TiO2/collagen-chitosan porous scaffold for wound repairing. Int J Biol Macromol. 2016;91:15-22.

[25]Yan F, Yue W, Zhang YL, et al. Chitosan-collagen porous scaffold and bone marrow mesenchymal stem cell transplantation for ischemic stroke. Neural Regen Res. 2015;10(9):1421-1426.

[26]Haaparanta AM, Järvinen E, Cengiz IF, et al. Preparation and characterization of collagen/PLA, chitosan/PLA, and collagen/chitosan/PLA hybrid scaffolds for cartilage tissue engineering. J Mater Sci Mater Med. 2014;25(4):1129-1136.

[27]Han J, Zhou Z, Yin R, et al. Alginate-chitosan/hydroxyapatite polyelectrolyte complex porous scaffolds: preparation and characterization. Int J Biol Macromol. 2010;46(2):199-205.

[28]Mathews S, Bhonde R, Gupta PK, et al. Novel biomimetic tripolymer scaffolds consisting of chitosan, collagen type 1, and hyaluronic acid for bone marrow-derived human mesenchymal stem cells-based bone tissue engineering. J Biomed Mater Res B Appl Biomater. 2014;102(8):1825-1834.

[29]Pati F, Adhikari B, Dhara S. Collagen intermingled chitosan-tripolyphosphate nano/micro fibrous scaffolds for tissue-engineering application. J Biomater Sci Polym Ed. 2012;23(15):1923-1938.

[30]Wang A, Ao Q, Cao W, et al. Porous chitosan tubular scaffolds with knitted outer wall and controllable inner structure for nerve tissue engineering. J Biomed Mater Res A. 2006;79(1):36-46.

[31]Wang X, Li Q, Hu X, et al. Fabrication and characterization of poly(L-lactide-co-glycolide) knitted mesh-reinforced collagen-chitosan hybrid scaffolds for dermal tissue engineering. J Mech Behav Biomed Mater. 2012;8:204-215.

[32]Bi L, Cao Z, Hu Y, et al. Effects of different cross-linking conditions on the properties of genipin-cross-linked chitosan/collagen scaffolds for cartilage tissue engineering. J Mater Sci Mater Med. 2011;22(1):51-62.

[33]Zhu Y, Liu T, Song K, et al. Ex vivo expansion of adipose tissue-derived stem cells in spinner flasks. Biotechnol J. 2009;4(8):1198-1209.

[34]Adekogbe I, Ghanem A. Fabrication and characterization of DTBP-crosslinked chitosan scaffolds for skin tissue engineering. Biomaterials. 2005;26(35):7241-7250.

[35]Ma F, Zhu T, Xu F, et al. Neural stem/progenitor cells on collagen with anchored basic fibroblast growth factor as potential natural nerve conduits for facial nerve regeneration. Acta Biomater. 2017;50:188-197.

[36]Li Q, Lu J, Wang X. Propofol and remifentanil at moderate and high concentrations affect proliferation and differentiation of neural stem/progenitor cells. Neural Regen Res. 2014; 9(22):2002-2007.

[37]Lu J, Li D, Lu K. Distribution and localization of fibroblast growth factor-8 in rat brain and nerve cells during neural stem/progenitor cell differentiation. Neural Regen Res. 2012;7(19):1455-1462.

[38]Lu J, Lu K, Li D. Changes in expression and secretion patterns of fibroblast growth factor 8 and Sonic Hedgehog signaling pathway molecules during murine neural stem/progenitor cell differentiation in vitro. Neural Regen Res. 2012;7(22):1688-1694.

[39]Kleiderman S, Gutbier S, Ugur Tufekci K, et al. Conversion of Nonproliferating Astrocytes into Neurogenic Neural Stem Cells: Control by FGF2 and Interferon-γ. Stem Cells. 2016; 34(12):2861-2874.

[40]Porseva VV, Shilkin VV, Korzina MB, et al. Peculiarities of age changes of NF200(+)-neurons in the sensory ganglia of different segmental levels after chemical deafferentation. Morfologiia. 2012;142(4):37-42.

[41]Johnson MD, Glick AD, Davis BW. Immunohistochemical evaluation of Leu-7, myelin basic-protein, S100-protein, glial-fibrillary acidic-protein, and LN3 immunoreactivity in nerve sheath tumors and sarcomas. Arch Pathol Lab Med. 1988;112(2):155-160.

[42]Tiainen H, Lyngstadaas SP, Ellingsen JE, et al. Ultra-porous titanium oxide scaffold with high compressive strength. J Mater Sci Mater Med. 2010;21(10):2783-2792.

[43]Wang Y, Wang X, Shang J, et al. Repairing the ruptured annular fibrosus by using type I collagen combined with citric acid, EDC and NHS: an in vivo study. Eur Spine J. 2017;26(3): 884-893.

[44]Liu EY, Jung S, Yi H. Improved Protein Conjugation with Uniform, Macroporous Poly(Acrylamide-co-Acrylic Acid) Hydrogel Microspheres via EDC/NHS Chemistry. Langmuir. 2016. in press.

[45]Hua J, Li Z, Xia W, et al. Preparation and properties of EDC/NHS mediated crosslinking poly (gamma-glutamic acid)/epsilon-polylysine hydrogels. Mater Sci Eng C Mater Biol Appl. 2016;61:879-892.

[46]Raghav R, Srivastava S. Immobilization strategy for enhancing sensitivity of immunosensors: L-Asparagine-AuNPs as a promising alternative of EDC-NHS activated citrate-AuNPs for antibody immobilization. Biosens Bioelectron. 2016;78:396-403.

[47]Park C, Vo CL, Kang T, et al. New method and characterization of self-assembled gelatin-oleic nanoparticles using a desolvation method via carbodiimide/ N-hydroxysuccinimide (EDC/NHS) reaction. Eur J Pharm Biopharm. 2015;89:365-373.

[48]Shepherd DV, Shepherd JH, Ghose S, et al. The process of EDC-NHS Cross-linking of reconstituted collagen fibres increases collagen fibrillar order and alignment. APL Mater. 2015;3(1). pii: 014902.

[49]Krishnamoorthy G, Selvakumar R, Sastry TP, et al. Experimental and theoretical studies on Gallic acid assisted EDC/NHS initiated crosslinked collagen scaffolds. Mater Sci Eng C Mater Biol Appl. 2014;43:164-171.

[50]Hao Z, Song Z, Huang J, et al. The scaffold microenvironment for stem cell based bone tissue engineering. Biomater Sci. 2017. in press.

[51]Sadtler K, Allen BW, Estrellas K, et al. The Scaffold Immune Microenvironment: Biomaterial-Mediated Immune Polarization in Traumatic and Nontraumatic Applications. Tissue Eng Part A. 2016. in press.

[52]Peng CH, Chuang JH, Wang ML, et al. Laminin modification subretinal bio-scaffold remodels retinal pigment epithelium-driven microenvironment in vitro and in vivo. Oncotarget. 2016;7(40):64631-64648.

[53]Sadtler K, Estrellas K, Allen BW, et al. Developing a pro-regenerative biomaterial scaffold microenvironment requires T helper 2 cells. Science. 2016;352(6283):366-370.

[54]Badylak SF. Tissue regeneration. A scaffold immune microenvironment. Science. 2016;352(6283):298.

[55]Kim J, Ma T. Perfusion regulation of hMSC microenvironment and osteogenic differentiation in 3D scaffold. Biotechnol Bioeng. 2012;109(1):252-261.

[56]Vrana NE, Builles N, Kocak H, et al. EDC/NHS cross-linked collagen foams as scaffolds for artificial corneal stroma. J Biomater Sci Polym Ed. 2007;18(12):1527-1545.

[57]Chang JY, Lin JH, Yao CH, et al. In vivo evaluation of a biodegradable EDC/NHS-cross-linked gelatin peripheral nerve guide conduit material. Macromol Biosci. 2007;7(4): 500-507.

[58]Krishnamoorthy G, Sehgal PK, Mandal AB, et al. Novel collagen scaffolds prepared by using unnatural D-amino acids assisted EDC/NHS crosslinking. J Biomater Sci Polym Ed. 2013;24(3):344-364.