Biological effects different diameters of Tussah silk fibroin nanofibers on olfactory ensheathing cells

Wu,Peng; Zhang,Peng; Zheng,Hanjiang; Zuo,Baoqi; Duan,Xiaofeng; Chen,Junjun; Wang,Xinhong; Shen,Yixin

doi:10.3892/etm.2018.6933

Biological effects different diameters of Tussah silk fibroin nanofibers on olfactory ensheathing cells

Authors:
- Peng Wu
- Peng Zhang
- Hanjiang Zheng
- Baoqi Zuo
- Xiaofeng Duan
- Junjun Chen
- Xinhong Wang
- Yixin Shen
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Affiliations: Department of Orthopedics, The Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215004, P.R. China, Department of Orthopedics, The Second Hospital of Jingzhou, Jingzhou, Hubei 434000, P.R. China, National Engineering Laboratory for Modern Silk, College of Textile and Clothing Engineering, Soochow University, Suzhou, Jiangsu 215004, P.R. China
Published online on: November 6, 2018 https://doi.org/10.3892/etm.2018.6933
Pages: 123-130
Copyright: © Wu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Transplantation of olfactory ensheathing cells (OECs) has potential for treating spinal cord and brain injury. However, they are void of an extracellular matrix to support cell growth and migration. Engineering of tissue to mimic the extracellular matrix is a potential solution for neural repair. Tussah silk fibroin (TSF) has good biocompatibility and an Arg‑Gly‑Asp tripeptide sequence. A small number of studies have assessed the effect of the diameter of TSF nanofibers on cell adhesion, growth and migration. In the present study, TSF nanofibers with a diameter of 400 and 1,200 nm were prepared using electrospinning technology; these were then used as scaffolds for OECs. The structure and morphology of the TSF nanofibers were characterized by scanning electron microscopy (SEM) and Fourier‑transform infrared spectroscopy. An inverted‑phase contrast microscope and SEM were used to observe the morphology of OECs on the TSF nanofibers. The effect on the adhesion of the cells was observed following crystal violet staining. The phenotype of the cells and the maximum axon length on the scaffolds were evaluated by immunostaining for nerve growth factor receptor p75. Cell proliferation and viability were assessed by an MTT assay and a Live/Dead reagent kit. The migration efficiency of OECs was observed using live‑cell microscopy. The results indicated that a 400‑nm TSF fiber scaffold was more conducive to OEC adhesion, growth and migration compared with a 1,200‑nm TSF scaffold. The phenotype of the OECs was normal, and no difference in OEC phenotype was observe when comparing those on TSF nanofibers to those on PLL. The present study may provide guidance regarding the preparation of tissue‑engineered materials for neural repair.

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1	Voronova АD, Stepanova OV, Valikhov MP, Chadin AV, Dvornikov АS, Reshetov IV and Chekhonin VP: Preparation of human olfactory ensheathing cells for the therapy of spinal cord injuries. Bull Exp Biol Med. 164:523–527. 2018. View Article : Google Scholar : PubMed/NCBI
2	Straley KS, Foo CW and Heilshorn SC: Biomaterial design strategies for the treatment of spinal cord injuries. J Neurotrauma. 27:1–19. 2010. View Article : Google Scholar : PubMed/NCBI
3	Gu M, Gao Z, Li X, Zhao F, Guo L, Liu J and He X: Feasibility of diffusion tensor imaging for assessing functional recovery in rats with olfactory ensheathing cell transplantation after contusive spinal cord injury (SCI). Med Sci Monit. 23:2961–2971. 2017. View Article : Google Scholar : PubMed/NCBI
4	Gu M, Gao Z, Li X, Guo L, Lu T, Li Y and He X: Conditioned medium of olfactory ensheathing cells promotes the functional recovery and axonal regeneration after contusive spinal cord injury. Brain Res. 1654:43–54. 2017. View Article : Google Scholar : PubMed/NCBI
5	Au E and Roskams AJ: Olfactory ensheathing cells of the lamina propria in vivo and in vitro. Glia. 41:224–236. 2003. View Article : Google Scholar : PubMed/NCBI
6	Khankan RR, Griffis KG, Haggerty-Skeans JR, Zhong H, Roy RR, Edgerton VR and Phelps PE: Olfactory ensheathing cell transplantation after a complete spinal cord transection mediates neuroprotective and immunomodulatory mechanisms to facilitate regeneration. J Neurosci. 36:6269–6286. 2016. View Article : Google Scholar : PubMed/NCBI
7	Führmann T, Anandakumaran PN and Shoichet MS: Combinatorial therapies after spinal cord injury: How can biomaterials help? Adv Healthc Mater. 6:2017.(doi: 10.1002/adhm.201601130). View Article : Google Scholar
8	Zhang Q, Yan S, You R, Kaplan DL, Liu Y, Qu J, Li X, Li M and Wang X: Multichannel silk protein/laminin grafts for spinal cord injury repair. J Biomed Mater Res A. 104:3045–3057. 2016. View Article : Google Scholar : PubMed/NCBI
9	Zhu S, Ge J, Wang Y, Qi F, Ma T, Wang M, Yang Y, Liu Z, Huang J and Luo Z: A synthetic oxygen carrier-olfactory ensheathing cell composition system for the promotion of sciatic nerve regeneration. Biomaterials. 35:1450–1461. 2014. View Article : Google Scholar : PubMed/NCBI
10	Kabiri M, Oraee-Yazdani S, Dodel M, Hanaee-Ahvaz H, Soudi S, Seyedjafari E, Salehi M and Soleimani M: Cytocompatibility of a conductive nanofibrous carbon nanotube/poly (L-Lactic acid) composite scaffold intended for nerve tissue engineering. EXCLI J. 14:851–860. 2015.PubMed/NCBI
11	Zhou M, Qiao W, Liu Z, Shang T, Qiao T, Mao C and Liu C: Development and in vivo evaluation of small-diameter vascular grafts engineered by outgrowth endothelial cells and electrospun chitosan/poly(ε-caprolactone) nanofibrous scaffolds. Tissue Eng Part A. 20:79–91. 2014. View Article : Google Scholar : PubMed/NCBI
12	Fan Z, Shen Y, Zhang F, Zuo B, Lu Q, Wu P, Xie Z, Dong Q and Zhang H: Control of olfactory ensheathing cell behaviors by electrospun silk fibroin fibers. Cell Transplant. 22 Suppl 1:S39–S50. 2013. View Article : Google Scholar : PubMed/NCBI
13	Sofia S, McCarthy MB, Gronowicz G and Kaplan DL: Functionalized silk-based biomaterials for bone formation. J Biomed Mater Res. 54:139–148. 2001. View Article : Google Scholar : PubMed/NCBI
14	Pavoni E, Tozzi S, Tsukada M and Taddei P: Structural study on methacrylamide-grafted Tussah silk fibroin fibres. Int J Biol Macromol. 88:196–205. 2016. View Article : Google Scholar : PubMed/NCBI
15	Gao Y, Shao W, Qian W, He J, Zhou Y, Qi K, Wang L, Cui S and Wang R: Biomineralized poly (l-lactic-co-glycolic acid)-tussah silk fibroin nanofiber fabric with hierarchical architecture as a scaffold for bone tissue engineering. Mater Sci Eng C Mater Biol Appl. 84:195–207. 2018. View Article : Google Scholar : PubMed/NCBI
16	Asakura T, Nishi H, Nagano A, Yoshida A, Nakazawa Y, Kamiya M and Demura M: NMR analysis of the fibronectin cell-adhesive sequence, Arg-Gly-Asp, in a recombinant silk-like protein and a model peptide. Biomacromolecules. 12:3910–3916. 2011. View Article : Google Scholar : PubMed/NCBI
17	Min BM, Lee G, Kim SH, Nam YS, Lee TS and Park WH: Electrospinning of silk fibroin nanofibers and its effect on the adhesion and spreading of normal human keratinocytes and fibroblasts in vitro. Biomaterials. 25:1289–1297. 2004. View Article : Google Scholar : PubMed/NCBI
18	Yang F, Murugan R, Wang S and Ramakrishna S: Electrospinning of nano/micro scale poly(L-lactic acid) aligned fibers and their potential in neuronal tissue engineering. Biomaterials. 26:2603–2610. 2005. View Article : Google Scholar : PubMed/NCBI
19	Smeal RM, Rabbitt R, Biran R and Tresco PA: Substrate curvature influences the direction of nerve outgrowth. Ann Biomed Eng. 33:376–382. 2005. View Article : Google Scholar : PubMed/NCBI
20	Smeal RM and Tresco PA: The influence of substrate curvature on neurite outgrowth is cell type dependent. Exp Neurol. 213:281–292. 2008. View Article : Google Scholar : PubMed/NCBI
21	Qu J, Wang D, Wang H, Dong Y, Zhang F, Zuo B and Zhang H: Electrospun silk fibroin nanofibers in different diameters support neurite outgrowth and promote astrocyte migration. J Biomed Mater Res A. 101:2667–2678. 2013. View Article : Google Scholar : PubMed/NCBI
22	Wang J, Ye R, Wei Y, Wang H, Xu X, Zhang F, Qu J, Zuo B and Zhang H: The effects of electrospun TSF nanofiber diameter and alignment on neuronal differentiation of human embryonic stem cells. J Biomed Mater Res A. 100:632–645. 2012. View Article : Google Scholar : PubMed/NCBI
23	Panda N, Bissoyi A, Pramanik K and Biswas A: Development of novel electrospun nanofibrous scaffold from P. Ricini and A. Mylitta silk fibroin blend with improved surface and biological properties. Mater Sci Eng C Mater Biol Appl. 48:521–532. 2015. View Article : Google Scholar : PubMed/NCBI
24	Khamhaengpol A and Siri S: Composite electrospun scaffold derived from recombinant fibroin of weaver ant (Oecophylla smaragdina) as cell-substratum. Appl Biochem Biotechnol. 183:110–125. 2010. View Article : Google Scholar
25	Yang R, Wu P, Wang X, Liu Z, Zhang C, Shi Y, Zhang F and Zuo B: A novel method to prepare tussah/Bombyx mori silk fibroin-based films. RSC Adv. 8:22069–22077. 2018. View Article : Google Scholar
26	Parker J: The protection of laboratory animals: A response to Stephenson. J Med Philos. 19:389–394. 1994. View Article : Google Scholar : PubMed/NCBI
27	Min BM, Jeong L, Nam YS, Kim JM, Kim JY and Park WH: Formation of silk fibroin matrices with different texture and its cellular response to normal human keratinocytes. Int J Biol Macromol. 34:281–288. 2004. View Article : Google Scholar : PubMed/NCBI
28	Zhuang Y, Zhang Q, Feng J, Wang N, Xu W and Yang H: The effect of native silk fibroin powder on the physical properties and biocompatibility of biomedical polyurethane membrane. Proc Inst Mech Eng H. 231:337–346. 2017. View Article : Google Scholar : PubMed/NCBI
29	Qi Y, Wang H, Wei K, Yang Y, Zheng RY, Kim IS and Zhang KQ: A review of structure construction of silk fibroin biomaterials from single structures to multi-level structures. Int J Mol Sci. 18:pii: E237. 2017. View Article : Google Scholar
30	Zhang W, Chen L, Chen J, Wang L, Gui X, Ran J, Xu G, Zhao H, Zeng M, Ji J, et al: Silk fibroin biomaterial shows safe and effective wound healing in animal models and a randomized controlled clinical trial. Adv Healthc Mater. 6:2017.(doi: 10.1002/adhm.201700121). View Article : Google Scholar
31	Fernández-García L, Marí-Buyé N, Barios JA, Madurga R, Elices M, Pérez-Rigueiro J, Ramos M, Guinea GV and González-Nieto D: Safety and tolerability of silk fibroin hydrogels implanted into the mouse brain. Acta Biomater. 45:262–275. 2016. View Article : Google Scholar : PubMed/NCBI
32	Zhang Z, Yoo R, Wells M, Beebe TP Jr, Biran R and Tresco P: Neurite outgrowth on well-characterized surfaces: Preparation and characterization of chemically and spatially controlled fibronectin and RGD substrates with good bioactivity. Biomaterials. 26:47–61. 2005. View Article : Google Scholar : PubMed/NCBI
33	Luan XY, Wang Y, Duan X, Duan QY, Li MZ, Lu SZ, Zhang HX and Zhang XG: Attachment and growth of human bone marrow derived mesenchymal stem cells on regenerated antheraea pernyi silk fibroin films. Biomed Mater. 1:181–187. 2006. View Article : Google Scholar : PubMed/NCBI
34	Minoura N, Aiba S, Gotoh Y, Tsukada M and Imai Y: Attachment and growth of cultured fibroblast cells on silk protein matrices. J Biomed Mater Res. 29:1215–1221. 1995. View Article : Google Scholar : PubMed/NCBI
35	Shen Y, Qian Y, Zhang H, Zuo B, Lu Z, Fan Z, Zhang P, Zhang F and Zhou C: Guidance of olfactory ensheathing cell growth and migration on electrospun silk fibroin scaffolds. Cell Transplant. 19:147–157. 2010. View Article : Google Scholar : PubMed/NCBI
36	Aznar-Cervantes S, Pagán A, Martínez JG, Bernabeu-Esclapez A, Otero TF, Meseguer-Olmo L, Paredes JI and Cenis JL: Electrospun silk fibroin scaffolds coated with reduced graphene promote neurite outgrowth of PC-12 cells under electrical stimulation. Mater Sci Eng C Mater Biol Appl. 79:315–325. 2017. View Article : Google Scholar : PubMed/NCBI