Characterization of chitosan-modified polyetheretherketone and its effect on MC3T3-E1 cell adhesion and proliferation

doi:10.12307/2022.643

Abstract

Abstract: BACKGROUND: The biological inert surface of polyetheretherketone limits its medical application, so how to improve the biological activity of polyetheretherketone needs to be solved urgently.
OBJECTIVE: To analyze the surface characteristics of polyetheretherketone modified by chitosan bioactive coating and its effect on the adhesion and proliferation of MC3T3-E1 cells.
METHODS: Disc-shaped polyetheretherketone material was obtained and treated with NaBH4, 3-aminopropyltriethoxysilane, glutaraldehyde aqueous solution, and chitosan solution to form polyetheretherketone modified with chitosan bioactive coating. X-ray photoelectron spectroscopy, scanning electron microscope, atomic force microscope, and automatic contact angle measurement instrument were used to observe the surface characteristics of chemically treated polyetheretherketone. MC3T3-E1 cells were seeded on the surface of polyetheretherketone and chitosan-modified polyetheretherketone separately, and the proliferation and adhesion of the cells were observed.
RESULTS AND CONCLUSION: (1) X-ray photoelectron spectroscopy showed that the polyetheretherketone material only contained C and O elements. The chitosan modified polyetheretherketone material contained C, O, N, and Si elements. The contact angle on the surface of chitosan modified polyetheretherketone material was smaller than that of the polyetheretherketone material (P < 0.05). (2) Under the scanning electron microscope, the surface of the polyetheretherketone material had obvious groove-like undulations, and the surface of the chitosan-modified polyetheretherketone material had chitosan molecule. The size was 1.0-2.0 μm. Atomic force microscope exhibited that there were many tiny pits on the surface of the polyetheretherketone material, and the size was about 0.1 μm. The pits on the surface of the chitosan-modified polyetheretherketone material increased; the size was 0.2-0.5 μm, and the surface roughness was greater than that of the polyetheretherketone material. (3) Under an inverted microscope, the number of cells on the surface of the chitosan-modified polyetheretherketone material was more than that of the polyetheretherketone material (P < 0.05 ). Under a laser confocal microscope, the cells adhered to the surface of the polyetheretherketone material had poor stretchability, fewer pseudopods, and inconspicuous actin microfilaments. The extensibility of cells adhered to the surface of the chitosan-modified polyetheretherketone material was better; the pseudopodia were more; the actin microfilament was more and obvious. (4) CCK-8 experiment showed that the cell proliferation on the surface of the chitosan-modified polyetheretherketone material was faster than that of the polyetheretherketone material (P < 0.05). (5) These results confirm that chitosan surface modification increases surface roughness and wettability of polyetheretherketone, promotes adhesion and proliferation of MC3T3-E1 cells on the surface of the material.

Key words: polyetheretherketone, chitosan, biomaterials, surface modification, biosafety, osteoblast, implant, cell proliferation, cell adhesion

CLC Number:

Feng Le, Qiu Peng, Liu Min, Zhou Hui. Characterization of chitosan-modified polyetheretherketone and its effect on MC3T3-E1 cell adhesion and proliferation[J]. Chinese Journal of Tissue Engineering Research, 2022, 26(21): 3351-3356.

Figures/Tables 9

References

[1] 陈贤明,周王峰.聚醚醚酮在口腔医学领域的应用[J].中国标准化,2020 (S01):354-358.
[2] PANAYOTOV IV, ORTI V, CUISINIER F, et al. Polyetheretherketone (PEEK) for medical applications. J Mater Sci Mater Med. 2016;27(7):118.
[3] TSUNEO S, MIYUKI H. Radiation deterioration of several aromatic polymers under oxidative conditions. Elsevier. 1987;28(11):694-699.
[4] LI HM, FOURACRE RA, GIVEN MJ, et al. The effects on polyetheretherketone and polyethersulfone of electron and /spl gamma/irradiation. IEEE T DIELECT EL IN. 1999;6(3):295-303.
[5] 陈井春雨,蓝菁.聚醚醚酮材料表面形态和骨结合原理研究进展[J].实用口腔医学杂志,2021,37(2):259-262.
[6] ABDULLAH MR, GOHARIAN A, ABDUL KADIR MR, et al. Biomechanical and bioactivity concepts of polyetheretherketone composites for use in orthopedic implants-a review. J Biomed Mater Res A. 2015;103(11):3689-3702.
[7] 刘吕花,熊成东,张丽芳,等.聚醚醚酮表面接枝聚丙烯酸及其成骨细胞相容性[J].塑料工业,2021,49(7):1591-1563.
[8] SAROT JR, CONTAR CM, CRUZ AC, et al. Evaluation of the stress distribution in CFR-PEEK dental implants by the three-dimensional finite element method. J Mater Sci Mater Med. 2010;21(7):2079-2085.
[9] FEERICK EM, KENNEDY J, MULLETT H, et al. Investigation of metallic and carbon fibre PEEK fracture fixation devices for three-part proximal humeral fractures. Med Eng Phys. 2013;35(6):712-722.
[10] OLIVARES-NAVARRETE R, GITTENS RA, SCHNEIDER JM, et al. Osteoblasts exhibit a more differentiated phenotype and increased bone morphogenetic protein production on titanium alloy substrates than on poly-ether-ether-ketone. Spine J. 2012;12(3):265-272.
[11] OLIVARES-NAVARRETE R, HYZY SL, SLOSAR PJ, et al. Implant materials generate different peri-implant inflammatory factors: poly-ether-ether-ketone promotes fibrosis and microtextured titanium promotes osteogenic factors. Spine (Phila Pa 1976). 2015;40(6):399-404.
[12] MUXIKA A, ETXABIDE A, URANGA J, et al. Chitosan as a bioactive polymer: Processing, properties and applications. Int J Biol Macromol. 2017;105(Pt 2): 1358-1368.
[13] PATRULEA V, OSTAFE V, BORCHARD G, et al. Chitosan as a starting material for wound healing applications. Eur J Pharm Biopharm. 2015;97(Pt B):417-426.
[14] 毛誉蓉,孙佳敏,周雄,等.医用特种高分子聚醚醚酮植入体及其表面界面工程[J].功能高分子学报,2021,34(2):144-160.
[15] MA R, TANG T. Current strategies to improve the bioactivity of PEEK. Int J Mol Sci. 2014;15(4):5426-5445.
[16] ABDULKAREEM MH, ABDALSALAM AH, BOHAN AJ. Influence of chitosan on the antibacterial activity of composite coating (PEEK /HAp) fabricated by electrophoretic deposition. Prog Org Coat. 2019;130:251-259.
[17] 陈睿超,孙辉,许国志.PEEK的表面改性和表征[J].中国塑料,2011,25(5): 17-23.
[18] 刘焕萍,孙辉,高艺菡.聚醚醚酮表面壳聚糖的固定[J].中国塑料,2014, 28(6):52-56.
[19] XU A, ZHOU L, DENG Y, et al. A carboxymethyl chitosan and peptide-decorated polyetheretherketone ternary biocomposite with enhanced antibacterial activity and osseointegration as orthopedic/dental implants. J Mater Chem B. 2016;4(10):1878-1890.
[20] BOLYEN E, RIDEOUT JR, DILLON MR, et al. Reproducible, interactive, scalable and extensible microbiome data science using QIIME 2. Nat Biotechnol. 2019;37(8):852-857.
[21] AZZAWI ZGM, HAMAD TI, KADHIM SA, et al. Osseointegration evaluation of laser-deposited titanium dioxide nanoparticles on commercially pure titanium dental implants. J Mater Sci Mater Med. 2018;29(7):96.
[22] GUADARRAMA BELLO D, FOUILLEN A, BADIA A, et al. A nanoporous titanium surface promotes the maturation of focal adhesions and formation of filopodia with distinctive nanoscale protrusions by osteogenic cells. Acta Biomater. 2017;60:339-349.
[23] FARAWATI FA, NAKAPARKSIN P. What is the Optimal Material for Implant Prosthesis? Dent Clin North Am. 2019;63(3):515-530.
[24] 鲁晨,陶璐,项闫颜,等.聚醚醚酮改性及在口腔种植领域的应用研究[J].实用口腔医学杂志,2020,36(5):819-824.
[25] 潘硕,郭晓恒.聚醚醚酮在口腔种植体及义齿修复领域的研究进展[J].世界最新医学信息文摘, 2019,19(84):40-42+44.
[26] MASSIA SP, STARK J. Immobilized RGD peptides on surface-grafted dextran promote biospecific cell attachment. J Biomed Mater Res. 2001;56(3):390-309.
[27] 陈兰花,盛道鹏.X射线光电子能谱分析(XPS)表征技术研究及其应用[J].教育现代化,2018,5(1):180-182+92.
[28] LIEDER R, DARAI M, THOR MB, et al. In vitro bioactivity of different degree of deacetylation chitosan, a potential coating material for titanium implants. J Biomed Mater Res A. 2012;100(12):3392-3399.
[29] GALLI C, PIEMONTESE M, LUMETTI S, et al. The importance of WNT pathways for bone metabolism and their regulation by implant topography. Eur Cell Mater. 2012;24:46-59.
[30] LV L, LIU Y, ZHANG P, et al. The nanoscale geometry of TiO2 nanotubes influences the osteogenic differentiation of human adipose-derived stem cells by modulating H3K4 trimethylation. Biomaterials. 2015;39:193-205.
[31] BAN S, IWAYA Y, KONO H, et al. Surface modification of titanium by etching in concentrated sulfuric acid. Dent Mater. 2006;22(12):1115-1120.
[32] DE SOUZA FERREIRA SB, DE ASSIS DIAS BR, OBREGON CS, et al. Microparticles containing propolis and metronidazole: in vitro characterization, release study and antimicrobial activity against periodontal pathogens. Pharm Dev Technol. 2014;19(2):173-180.
[33] MAMALIS AA, MARKOPOULOU C, VROTSOS I, et al. Chemical modification of an implant surface increases osteogenesis and simultaneously reduces osteoclastogenesis: an in vitro study. Clin Oral Implants Res. 2011;22(6): 619-626.
[34] BANG SM, MOON HJ, KWON YD, et al. Osteoblastic and osteoclastic differentiation on SLA and hydrophilic modified SLA titanium surfaces. Clin Oral Implants Res. 2014;25(7):831-837.
[35] 李雪菁,邱小亥,赵宝红.亲水种植体促进骨整合机制研究进展[J].中国实用口腔科杂志,2020,13(8):501-505.
[36] CONTI M, DONATI G, CIANCIOLO G, et al. Force spectroscopy study of the adhesion of plasma proteins to the surface of a dialysis membrane: role of the nanoscale surface hydrophobicity and topography. J Biomed Mater Res. 2002;61(3):370-379.
[37] AMARAL IF, LAMGHARI M, SOUSA SR, et al. Rat bone marrow stromal cell osteogenic differentiation and fibronectin adsorption on chitosan membranes: the effect of the degree of acetylation. J Biomed Mater Res A. 2005;75(2):387-397.
[38] MOURSI AM, GLOBUS RK, DAMSKY CH. Interactions between integrin receptors and fibronectin are required for calvarial osteoblast differentiation in vitro. J Cell Sci. 1997;110(Pt 18):2187-2196.
[39] SAENGDEE P, CHAISRIRATANAKUL W, BUNJONGPRU W, et al. Surface modification of silicon dioxide, silicon nitride and titanium oxynitride for lactate dehydrogenase immobilization. Biosens Bioelectron. 2015;67:134-138.
[40] ZHU X, CHEN J, SCHEIDELER L, et al. Effects of topography and composition of titanium surface oxides on osteoblast responses. Biomaterials. 2004; 25(18):4087-4103.
[41] ANSELME K. Osteoblast adhesion on biomaterials. Biomaterials. 2000;21(7): 667-681.
[42] MARTIN HJ, SCHULZ KH, BUMGARDNER JD, et al. XPS study on the use of 3-aminopropyltriethoxysilane to bond chitosan to a titanium surface. Langmuir. 2007;23(12):6645-6651.
[43] HSIAO SW, THIEN DV, HO MH, et al. Interactions between chitosan and cells measured by AFM. Biomed Mater. 2010;5(5):054117.
[44] CAI K, FRANT M, BOSSERT J, et al. Surface functionalized titanium thin films: zeta-potential, protein adsorption and cell proliferation. Colloids Surf B Biointerfaces. 2006;50(1):1-8.
[45] 俞娟,徐俊华,范一民.壳聚糖抗菌性能研究进展[J].林业工程学报, 2018,3(5):20-27.