Synthesis and evaluation of cytotoxicity of hybrid hydrogel based on chitosan and Ph.ZnO@HAP nanocomposite

Document Type : Original Research

Authors

1 Department of Materials & Polymer Engineering.Hakim Sabzevari University

2 Ferdowsi University of Mashhad, Faculty of Science, Department of Chemistry

Abstract
In this study, phenolic compounds-coated ZnO@HAP nanocomposite (Ph.ZnO@HAP) was synthesized and used to improve the physical and chemical properties of chitosan hydrogel for biological application. At first, the phenolic compounds were extracted from walnut green hulls. The synthesis of Ph.ZnO@HAP nanocomposite was performed with the assistance of extracted phenols using a hydrothermal method. Chitosan hydrogel was also prepared using NaHCO3 at 37°C. Hybrid hydrogels based on chitosan and Ph.ZnO@HAP nanocomposite were prepared in a similar way and then characterized by fourier-transform infrared spectroscopy (FTIR) and field emission scanning electron microscopy (FESEM). The antioxidant property, cytotoxicity, and osteogenesis of hybrid hydrogels were measured using DPPH radical scavenging method, MTT, and alkaline phosphatase enzyme assay, respectively. The FTIR spectra, FESEM images, EDX spectrum, and Zeta potential data showed that Ph.ZnO@HAP nanocomposites synthesized successfully with rod-like morphology, phenolic compounds coated on the surface and a negative particle surface charge. The results of DPPH experiment showed that the antioxidant property of the nanocomposite material increased in a concentration-dependent manner. The FESEM images of chitosan hybrid hydrogels with different concentrations of embedded Ph.ZnO@HAP nanocomposite showed that hybrid hydrogels have a more uniform porous structure, compared to the chitosan hydrogel. Moreover, by an increase in the nanocomposite concentration in the structure of hybrid hydrogels, the antioxidant property augmented. The results of the biological studies showed that the cytotoxicity of hybrid hydrogels on osteoblast-like cells (Saos-2) is lower than that of chitosan hydrogel. Also, hybrid hydrogels showed the higher potential in induction of osteogenesis than chitosan hydrogels.

Keywords

Subjects


1 Hamedi H., Moradi S., Hudson S.M., Tonelli AE. Chitosan based hydrogels and their applications for drug delivery in wound dressings: A review. Carbohydrate Polymers. 199, 445-60,2018.

2 El Knidri H., Belaabed R., Addaou A., Laajeb A., Lahsini A. Extraction, chemical modification and characterization of chitin and chitosan. International Journal of Biological Macromolecules. 120,1181-9, 2018.

3 Pellá M.C.G., Lima-Tenório M.K., Tenório-Neto E.T., Guilherme M.R., Muniz E.C., Rubira A.F. Chitosan-based hydrogels: From preparation to biomedical applications. Carbohydrate Polymers. 196, 233-45, 2018.

4 Ahmed S., Annu, Ali A., Sheikh J. A review on chitosan centred scaffolds and their applications in tissue engineering. International Journal of Biological Macromolecules. 116, 849-862, 2018.

5 Milosavljević N.B., Kljajević L.M., Popović I.G., Filipović J.M., Kalagasidis Krušić M.T. Chitosan, itaconic acid and poly(vinyl alcohol) hybrid polymer networks of high degree of swelling and good mechanical strength. Polymer International. 59(5),686-694, 2010.

6 Singh A., Narvi S.S., Dutta P.K., Pandey N.D. External stimuli response on a novel chitosan hydrogel crosslinked with formaldehyde. Bulletin of Materials Science. 29(3),233-8,2006.

7 Liu R., Xu X., Zhuang X., Cheng B. Solution blowing of chitosan/PVA hydrogel nanofiber mats. Carbohydr Polymer. 101,1116-21, 2014.

8 Muzzarelli R.A.A. Genipin-crosslinked chitosan hydrogels as biomedical and pharmaceutical aids. Carbohydrate Polymers. 77(1), 1-9, 2009.

9 Montembault A., Viton C., Domard A. Rheometric Study of the Gelation of Chitosan in Aqueous Solution without Cross-Linking Agent. Biomacromolecules. 6(2),653-62,2005.

10 Liu L., Tang X., Wang Y., Guo S. Smart gelation of chitosan solution in the presence of NaHCO3 for injectable drug delivery system. International journal of pharmaceutics. 414(1-2), 6-15, 2011.

11 Croisier F., Jérôme C. Chitosan-based biomaterials for tissue engineering. European Polymer Journal. 49(4), 780-92, 2013.
12 Xu B., Wang L., Liu Y., Zhu H., Wang Q. Preparation of high strength and transparent nanocomposite hydrogels using alumina nanoparticles as cross-linking agents. Materials Letters. 228, 104-7, 2018.

13 Xie Y., Liao X., Zhang J., Yang F., Fan Z. Novel chitosan hydrogels reinforced by silver nanoparticles with ultrahigh mechanical and high antibacterial properties for accelerating wound healing. International Journal of Biological Macromolecules. 119, 402-12, 2018.

14 Szcześ A., Hołysz L., Chibowski E. Synthesis of hydroxyapatite for biomedical applications. Advances in Colloid and Interface Science. 249, 321-30, 2017.

15 Yoshikawa H., Tamai N., Murase T., Myoui A. Interconnected porous hydroxyapatite ceramics for bone tissue engineering. Journal of the Royal Society, Interface. 6 Suppl 3(Suppl 3):S341-S8, 2009.

16 Nikpour M.R., Rabiee S.M., Jahanshahi M. Synthesis and characterization of hydroxyapatite/chitosan nanocomposite materials for medical engineering applications. Composites Part B: Engineering. 43(4),1881-6, 2012.

17 Chang C., Peng N., He M., Teramoto Y., Nishio Y., Zhang L. Fabrication and properties of chitin/hydroxyapatite hybrid hydrogels as scaffold nano-materials. Carbohydr Polymer. 91(1), 7-13, 2013.
.
18 Fricain J.C., Schlaubitz S., Le Visage C., Arnault I., Derkaoui S.M., Siadous R., et al. A nano-hydroxyapatite--pullulan/dextran polysaccharide composite macroporous material for bone tissue engineering. Biomaterials. 34(12), 2947-59, 2013.

19 Saxena V., Hasan A., Pandey L.M. Effect of Zn/ZnO integration with hydroxyapatite: a review. Materials Technology. 33(2), 79-92, 2018.
20Meshkini A. Modulation of Oxidative Stress in Thrombin-Stimulated Platelets by Almond by-Product. Waste and Biomass Valorization. 9(6), 1015-25, 2018.

21 Pezeshkpour V., Khosravani S.A., Ghaedi M., Dashtian K., Zare F., Sharifi A., et al. Ultrasound assisted extraction of phenolic acids from broccoli vegetable and using sonochemistry for preparation of MOF-5 nanocubes: Comparative study based on micro-dilution broth and plate count method for synergism antibacterial effect. Ultrasonics Sonochemistry. 40,1031-8, 2018.

22 Meshkini A. Acetone Extract of Almond Hulls Provides Protection against Oxidative Damage and Membrane Protein Degradation. Journal of acupuncture and meridian studies. 9(3), 134-42, 2016.

23 Bradford M.M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical biochemistry. 72, 248-54, 1976.
24Hashemian S. A Comparative Study of Cellulose Agricultural Wastes (Almond Shell, Pistachio Shell, Walnut Shell, Tea Waste And Orange Peel) for Adsorption of Violet B Dye from Aqueous Solutions. Oriental Journal Chemistry. 30, 4, 2014.

25 Zhang N., Gao T., Wang Y., Wang Z., Zhang P., Liu J. Environmental pH-controlled loading and release of protein on mesoporous hydroxyapatite nanoparticles for bone tissue engineering. Materials science & engineering C, Materials for biological applications. 46, 158-65, 2015.

26 Contri R.V., Soares R.M.D., Pohlmann A.R., Guterres S.S. Structural analysis of chitosan hydrogels containing polymeric nanocapsules. Materials Science and Engineering: C. 42, 234-42, 2014.

27 Ngo D.H., Kim S.K. Chapter Two - Antioxidant Effects of Chitin, Chitosan, and Their Derivatives. In: Kim S-K, editor. Advances in food and nutrition research. 73: Academic Press;. p. 15-31, 2014.
28 Ngo D.H., Kim S.K. Antioxidant effects of chitin, chitosan, and their derivatives. Advances in food and nutrition research. 73,15-31, 2014.

29 Jin P., Wu H., Xu G., Zheng L., Zhao J. Epigallocatechin-3-gallate (EGCG) as a pro-osteogenic agent to enhance osteogenic differentiation of mesenchymal stem cells from human bone marrow: an in vitro study. Cell and tissue research. 356(2), 381-90, 2014.

30 Sistanipour E., Meshkini A., Oveisi H. Catechin-conjugated mesoporous hydroxyapatite nanoparticle: A novel nano-antioxidant with enhanced osteogenic property. Colloids and surfaces B, Biointerface, 169, 329-339, 2018.

31 Aubin J.E. Regulation of osteoblast formation and function. Reviews in endocrine & metabolic disorders. 2(1), 81-94, 2001.

32 Rodriguez-Valencia C., Freixeiro P., Serra J., Ferreiros C.M., Gonzalez P., Lopez-Alvarez M. In vitro evaluation of the antibacterial and osteogenic activity promoted by selenium-doped calcium phosphate coatings. Biomedical materials (Bristol, England). 12(1), 015028, 2017.