Effects of nano-zirconium dioxide on osteogenic differentiation of ectomesenchymal stem cells in nasal mucosa

doi:10.12307/2024.409

Abstract

Abstract: BACKGROUND: Nano-zirconium dioxide has good application potential in the field of bone tissue repair. Studying the effect of nano-zirconium dioxide on osteogenic differentiation will help to promote the clinical application of nano-zirconium dioxide in the treatment of bone defects.
OBJECTIVE: To explore the effect of nano-zirconium dioxide on the osteogenic differentiation of ectomesenchymal stem cells in the nasal mucosa.
METHODS: Ectomesenchymal stem cells derived from rat nasal mucosa were isolated and cultured, and the biotoxicity of nano-zirconium dioxide to the cells was detected by CCK-8 assay. The biosafety concentration was selected according to the cytotoxicity, and the cells were randomly divided into a control group, a nano-zirconium dioxide group, and a nano-hydroxyapatite group. Osteogenic differentiation of cells was directionally induced in each group. On day 7 of induced differentiation, alkaline phosphatase staining was performed. qRT-PCR and western blot assay were used to detect the expression of early osteogenic markers (Runx2 and Osx). On day 21 of induced differentiation, alizarin red staining was conducted. qRT-PCR and western blot assay were utilized to determine the expression levels of late osteogenic markers (OPN and OCN).
RESULTS AND CONCLUSION: (1) The median lethal concentration of nano-zirconium dioxide on ectomesenchymal stem cells in nasal mucosa was 0.6 mg/mL. In the experiment, the mass concentration of 200 μg/mL was selected for intervention. Zirconium dioxide had no significant effect on the proliferation of the cells. (2) Compared with the control group, the alkaline phosphatase staining of the cells in the nano-zirconium dioxide group was more obvious and the level of cell mineralization was higher, but there was no significant difference compared with the nano-hydroxyapatite. (3) Compared with the control group, the expression of bone-related genes and proteins increased significantly, but there was no significant difference compared with nano-hydroxyapatite. (4) The results show that nano-zirconium dioxide has good biological safety and can promote the osteogenic differentiation of ectomesenchymal stem cells in the nasal mucosa. This promoting effect is equivalent to that of nano-hydroxyapatite.

Key words: nasal mucosa, ectomesenchymal stem cell, zirconium dioxide, hydroxyapatite, osteogenic differentiation

CLC Number:

Bian Lu, Xia Dandan, Qian Yuan, Shi Wen, Que Yunduan, Lyu Long, Xu Aihua, Shi Wentao. Effects of nano-zirconium dioxide on osteogenic differentiation of ectomesenchymal stem cells in nasal mucosa[J]. Chinese Journal of Tissue Engineering Research, 2024, 28(15): 2346-2350.

Figures/Tables 6

References

[1] 汪海平,刘芸,胡思前. ZrO2气凝胶材料的制备、改性及应用研究进展[J].江汉大学学报(自然科学版),2023,51(2):15-26.
[2] 徐臻,王斐,印霞,等. ZrO2纳米纤维的最新进展:从结构设计到新兴应用[J].中国科学:材料科学(英文版),2023,66(2):421-440.
[3] LINH NB, JANG DW, LEE BT. Collagen immobilization of multi-layered BCP-ZrO2 bone substitutes to enhance bone formation. Appl Surf Sci. 2015;345:238-248.
[4] HU C, SUN J, LONG C, et al. Synthesis of nano zirconium oxide and its application in dentistry. Nanotechnol Rev. 2019;8(1):396-404.
[5] WANG W, HOU FY, WANG H, et al. Fabrication and characterization of Ni-ZrO2 composite nano-coatings by pulse electrodeposition. Scr Mater. 2005;53(5):613-618.
[6] KONG YM, BAE CJ, LEE SH, et al. Improvement in biocompatibility of ZrO2-Al2O3 nano-composite by addition of HA. Biomaterials. 2005;26(5):509-517.
[7] YU W, WANG X, ZHAO J, et al. Preparation and mechanical properties of reinforced hydroxyapatite bone cement with nano-ZrO2. Ceram Int. 2015;41(9):10600-10606.
[8] WANG G, LIU X, ZREIQAT H, et al. Enhanced effects of nano-scale topography on the bioactivity and osteoblast behaviors of micron rough ZrO2 coatings. Colloid Surface B. 2011;86(2):267-274.
[9] PANG XF, HUANG Y, CAO XY. Preparation and Features of Nano-ZrO2/HA Coating on Surface of Titanium Materials in Dental-Implant. Adv Mater Res. 2011;295:491-495.
[10] LV M, LV W, CHEN H, et al. Biotribological properties of nano zirconium dioxide and hydroxyapatite-reinforced polyetheretherketone (HA/ZrO2/PEEK) biocomposites. Iran Polym J. 2021;30(11):1127-1136.
[11] SHI W, WANG Z, BIAN L, et al. Periodic Heat Stress Licenses EMSC Differentiation into Osteoblasts via YAP Signaling Pathway Activation. Stem Cells Int. 2022;2022:3715471.
[12] DENG W, SHAO F, HE Q, et al. EMSCs Build an All-in-One Niche via Cell-Cell Lipid Raft Assembly for Promoted Neuronal but Suppressed Astroglial Differentiation of Neural Stem Cells. Adv Mater. 2019;31(10):e1806861.
[13] CHEN Q, ZHANG Z, LIU J, et al. A fibrin matrix promotes the differentiation of EMSCs isolated from nasal respiratory mucosa to myelinating phenotypical Schwann-like cells. Mol Cells. 2015;38(3):221-228.
[14] SHI W, QUE Y, ZHANG X, et al. Functional tissue-engineered bone-like graft made of a fibrin scaffold and TG2 gene-modified EMSCs for bone defect repair. NPG Asia Mater. 2021;13(1):28.
[15] DHIVYA S, AJITA J, SELVAMURUGAN N. Metallic nanomaterials for bone tissue engineering. J Biomed Nanotechnol. 2015;11(10):1675-1700.
[16] LIU X, MIAO Y, LIANG H, et al. 3D-printed bioactive ceramic scaffolds with biomimetic micro/nano-HAp surfaces mediated cell fate and promoted bone augmentation of the bone-implant interface in vivo. Bioact Mater. 2021;12:120-132.
[17] DU M, CHEN J, LIU K, et al. Recent advances in biomedical engineering of nano-hydroxyapatite including dentistry, cancer treatment and bone repair. Compos Part B-Eng. 2021;215:108790.
[18] SHI W, WU Y, WU J, et al. NPS-Crosslinked Fibrin Gels Load with EMSCs to Repair Peripheral Nerve Injury in Rats. Macromol Biosci. 2023;23(3):e2200381.
[19] HAARHAUS M, CIANCIOLO G, BARBUTO S, et al. Alkaline Phosphatase: An Old Friend as Treatment Target for Cardiovascular and Mineral Bone Disorders in Chronic Kidney Disease. Nutrients. 2022;14(10):2124.
[20] JUNG JH, HONG CM, JO I, et al. Reliability of Alkaline Phosphatase for Differentiating Flare Phenomenon from Disease Progression with Bone Scintigraphy. Cancers (Basel). 2022;14(1):254.
[21] SHU J, TAN A, LI Y, et al. The correlation between serum total alkaline phosphatase and bone mineral density in young adults. BMC Musculoskelet Disord. 2022;23(1):467.
[22] YIN X, TENG X, MA T, et al. RUNX2 recruits the NuRD(MTA1)/CRL4B complex to promote breast cancer progression and bone metastasis. Cell Death Differ. 2022;29(11):2203-2217.
[23] NING K, YANG B, CHEN M, et al. Functional Heterogeneity of Bone Marrow Mesenchymal Stem Cell Subpopulations in Physiology and Pathology. Int J Mol Sci. 2022;23(19):11928.
[24] ABOU NEEL EA, KG AR, SAMSUDIN AR. Mineralized nodule formation in primary osteoblasts culture in titanium doped phosphate glass and in-house prepared freeze dried demineralized bone extracts. Mater Chem Phys. 2022;276:125425.
[25] LIN C, CHEN Z, GUO D, et al. Increased expression of osteopontin in subchondral bone promotes bone turnover and remodeling, and accelerates the progression of OA in a mouse model. Aging (Albany NY). 2022;14(1):253-271.
[26] SALAMA RHM, ALI SS, SALAMA THM, et al. Dietary Effects of Nanopowder Eggshells on Mineral Contents, Bone Turnover Biomarkers, and Regulators of Bone Resorption in Healthy Rats and Ovariectomy-Induced Osteoporosis Rat Model. Appl Biochem Biotechnol. 2023;195(8):5034-5052.
[27] SAFIAGHDAM H, NOKHBATOLFOGHAHAEI H, FARZAD-MOHAJERI S, et al. 3D-printed MgO nanoparticle loaded polycaprolactone β-tricalcium phosphate composite scaffold for bone tissue engineering applications: In-vitro and in-vivo evaluation. J Biomed Mater Res A. 2023;111(3):322-339.
[28] LIU S, LI K, SHEN Q, et al. Electroactive MnO2-poly (3, 4-ethylenedioxythiophene) composite nanocoatings enhance osteoblastic electrical stimulation. Appl Surf Sci. 2021;545:148827.
[29] WU Z, MENG Z, WU Q, et al. Biomimetic and osteogenic 3D silk fibroin composite scaffolds with nano MgO and mineralized hydroxyapatite for bone regeneration. J Tissue Eng. 2020; 11:2041731420967791.
[30] TARAFDER S, DAVIES NM, BANDYOPADHYAY A, et al. 3D printed tricalcium phosphate scaffolds: Effect of SrO and MgO doping on in vivo osteogenesis in a rat distal femoral defect model. Biomater Sci. 2013;1(12):1250-1259.
[31] SHAO RX, QUAN RF, WANG T, et al. Effects of a bone graft substitute consisting of porous gradient HA/ZrO2 and gelatin/chitosan slow-release hydrogel containing BMP-2 and BMSCs on lumbar vertebral defect repair in rhesus monkey. J Tissue Eng Regen Med. 2018;12(3): e1813-e1825.