Graphene oxide-chitosan composite coating affects the biological behavior of osteoblasts

doi:10.12307/2023.542

Abstract

Abstract: BACKGROUND: As a coating material, chitosan can effectively improve biological properties of medical metal surfaces due to its excellent film-forming and metal binding abilities. However, the biological activity and osteoinductive ability of chitosan are insufficient. Graphene oxide can improve the mechanical properties and biocompatibility of various polymer materials, as well as promote osteogenic differentiation. The combination of chitosan and graphene oxide may have better biological properties and osteogenic activity.
OBJECTIVE: To construct the graphene oxide-modified chitosan composite coating material, analyze the surface morphology, chemical composition, and physical properties of the coating material, as well as the effects of the coating materials on proliferation, adhesion, and osteogenic differentiation of osteoblasts.
METHODS: The titanium surface was treated with 3-aminopropyltriethoxysilane to form silane groups on the surface, and then glutaraldehyde was used to cross-link chitosan with silane groups to prepare simple chitosan coating and graphene oxide/chitosan composite coating, respectively. The surface morphology, chemical structure and hydrophilic properties of the coatings were characterized by atomic force microscopy, scanning electron microscopy, Fourier transform infrared spectroscopy, and contact angle measurement system. Rat osteoblasts were seeded on the surface of the two materials, and the proliferation, spreading, and osteogenic differentiation of rat osteoblasts were analyzed by CCK-8 assay, scanning electron microscopy, and reverse transcription-polymerase chain reaction analysis.
RESULTS AND CONCLUSION: (1) The thickness of the graphene oxide sheets was about 2 nm as measured by atomic force microscopy; the scanning electron microscopy showed that the surfaces of both coatings were smooth and dense, in which the chitosan molecules were closely arranged and the graphene oxide was uniformly distributed in the chitosan, while wrinkles appeared on the surface of chitosan due to the addition of grapheme. The contact angle of the graphene oxide-chitosan composite coating was significantly smaller than the chitosan coating alone (P < 0.05). (2) CCK-8 assay showed that the graphene oxide-chitosan composite coating better promoted the proliferation of osteoblasts compared with the chitosan coating alone. (3) The scanning electron microscopy showed that better spreading was found in osteoblasts on the surface of the graphene oxide-chitosan composite coating compared with the chitosan coating alone. (4) Reverse transcription-polymerase chain reaction analysis showed that the mRNA expression of alkaline phosphatase and Runx2 was significantly increased in the graphene oxide/chitosan composite coating group compared with in the chitosan coating alone group (P < 0.05). (5) These findings suggest that graphene oxide/chitosan composite coating had good biocompatibility, physicochemical, and osteoinductive properties.

Key words: graphene oxide, chitosan, two-dimensional material, coating material, hydrophilicity, cell adhesion, cell proliferation, osteogenic differentiation

CLC Number:

Huang Qian, Hao Liying, He Longlong, Du Liangzhi. Graphene oxide-chitosan composite coating affects the biological behavior of osteoblasts[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(34): 5430-5435.

Figures/Tables 8

References

[1] NOVOSELOV KS, GEIM AK, MOROZOV SV, et al. Electric field effect in atomically thin carbon films. Science. 2004;306(5696):666-669.
[2] NOVOSELOV KS, JIANG D, SCHEDIN F, et al. Two-dimensional atomic crystals. Proc Natl Acad Sci U S A. 2005;102(30):10451-10453.
[3] JIRICKOVA A, JANKOVSKY O, SOFER Z, et al. Synthesis and Applications of Graphene Oxide. Materials (Basel). 2022;15(3):920.
[4] JOUIBARY YM, REZVANPOUR A, AKBARI B. Fabrication of polycaprolactone/collagen scaffolds reinforced by graphene oxide for tissue engineering applications. Mater Lett. 2022;322.
[5] LI Q, YAN Y, GAO H. Improving the corrosion resistance and osteogenic differentiation of ZK60 magnesium alloys by hydroxyapatite/graphene/graphene oxide composite coating. Ceram Int. 2022;48(11):16131-16141.
[6] RAWAT S, JAIN KG, GUPTA D, et al. Graphene nanofiber composites for enhanced neuronal differentiation of human mesenchymal stem cells. Nanomedicine. 2021;16(22):1963-1982.
[7] TAO B, ZHAO W, LIN C, et al. Surface modification of titanium implants by ZIF-8@Levo/LBL coating for inhibition of bacterial-associated infection and enhancement of in vivo osseointegration. Chem Eng J. 2020;390:124621.
[8] YANG Z, XI Y, BAI J, et al. Covalent grafting of hyperbranched poly-L-lysine on Ti-based implants achieves dual functions of antibacteria and promoted osteointegration in vivo. Biomaterials. 2021;269:120534.
[9] FRANK LA, ONZI GR, MORAWSKI AS, et al. Chitosan as a coating material for nanoparticles intended for biomedical applications. React Funct Polym. 2020;147:104459.
[10] GARCIA-CABEZON C, GODINHO V, SALVO-COMINO C, et al. Improved Corrosion Behavior and Biocompatibility of Porous Titanium Samples Coated with Bioactive Chitosan-Based Nanocomposites. Materials (Basel). 2021;14(21):6322.
[11] HUNGE YM, YADAV AA, DHODAMANI AG, et al. Enhanced photocatalytic performance of ultrasound treated GO/TiO2 composite for photocatalytic degradation of salicylic acid under sunlight illumination. Ultrason Sonochem. 2020;61:104849.
[12] YU J, LIN M, TAN Q, et al. High-value utilization of graphite electrodes in spent lithium-ion batteries: From 3D waste graphite to 2D graphene oxide. J Hazard Mater. 2021;401:123715.
[13] 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.
[14] VILLEGAS M, ZHANG Y, BADV M, et al. Enhancing osseointegration and mitigating bacterial biofilms on medical-grade titanium with chitosan-conjugated liquid-infused coatings. Sci Rep. 2022;12(1):5380.
[15] ZHONG M, DAI X, XIANG H, et al. Preparation, Characterization, and Terahertz Spectroscopy Characteristics of Reduced Graphene Oxide-Doped Epoxy Resin Coating. Coatings. 2021;11(12). doi.org/10.3390/coatings11121503
[16] CHEN S, LI Q, HE D, et al. Aggregation behavior of partially contacted graphene sheets in six-carbon alkanes: all-atom molecular dynamics simulation. J Mol Model. 2022;28(6):149.
[17] SU L, SUN J, DING F, et al. Molecular insight into the reversible dispersion and aggregation of graphene utilizing photo-responsive surfactants. Appl Surf Sci. 2021;567:150840.
[18] TANG H, ZHANG S, HUANG T, et al. Mechanisms of the Aggregation of Graphene Oxide at High pH: Roles of Oxidation Debris and Metal Adsorption. Environ Sci Technol. 2021;55(21):14639-14648.
[19] PINELLI F, NESPOLI T, ROSSI F. Graphene Oxide-Chitosan Aerogels: Synthesis, Characterization, and Use as Adsorbent Material for Water Contaminants. Gels. 2021;7(4):149.
[20] YANG F, YUAN B, WANG Y, et al. Graphene oxide/chitosan nano-coating with ultrafast fire-alarm response and flame-retardant property. Polym Adv Technol. 2022;33(3):795-806.
[21] JUNG I, VAUPEL M, PELTON M, et al. Characterization of Thermally Reduced Graphene Oxide by Imaging Ellipsometry. J Phys Chem C. 2008;112(23):8499-8506.
[22] ZHENG J, DI CA, LIU Y, et al. High quality graphene with large flakes exfoliated by oleyl amine. Chem Commun. 2010;46(31):5728-5730.
[23] LI J, YAO Y, WANG Y, et al. Modulation of the Crosstalk between Schwann Cells and Macrophages for Nerve Regeneration: A Therapeutic Strategy Based on a Multifunctional Tetrahedral Framework Nucleic Acids System. Adv Mater. 2022;e2202513. doi: 10.1002/adma.202202513.
[24] TANG X, LI Q, LI L, et al. Expression of Talin-1 in endometriosis and its possible role in pathogenesis. Reprod Biol Endocrinol. 2021;19(1):42.
[25] MACDONALD AF, TROTTER RD, GRIFFIN CD, et al. Genetic profiling of human bone marrow and adipose tissue-derived mesenchymal stem cells reveals differences in osteogenic signaling mediated by graphene. J Nanobiotechnol. 2021;19(1):285.
[26] NEWBY SD, MASI T, GRIFFIN CD, et al. Functionalized Graphene Nanoparticles Induce Human Mesenchymal Stem Cells to Express Distinct Extracellular Matrix Proteins Mediating Osteogenesis. Int J Nanomed. 2020;15:2501-2513.
[27] EIVAZZADEH-KEIHAN R, MALEKI A, DE LA GUARDIA M, et al. Carbon based nanomaterials for tissue engineering of bone: Building new bone on small black scaffolds: A review. J Adv Res. 2019;18:185-201.
[28] LI H, HUANG J, WANG Y, et al. Nanoscale Modification of Titanium Implants Improves Behaviors of Bone Mesenchymal Stem Cells and Osteogenesis In Vivo. Oxid Med Cell Longevity. 2022;2022:2235335.