Evaluating Various Properties of Nanohydroxyapatite Synthesized from Eggshells and Dual-doped with Si and Zn: An in Vitro Study

Overview

Journal Heliyon

Specialty Social Sciences

Date 2024 Sep 3

PMID 39224256

Authors

Shaza Alashi

Isam Alkhouri

Ibrahim Alghoraibi

Nabil Kochaji

Abdullah Houri

Mawia Karkoutly

Affiliations

Soon will be listed here.

Abstract

Background: This study aimed to evaluate morphological, chemical and biocompatible properties of nanohydroxyapatite (N-HA) synthesized from eggshells and dual-doped with Si4+ and Zn2+.

Methods: In the current study, N-HA was synthesized from chicken eggshells using the wet chemical precipitation method and doped with Si4+ and Zn2+. The physical assessment was carried out using field emission scanning electron microscopy (FE-SEM), energy dispersive X-ray (EDX) analysis, and X-ray diffraction (XRD) analysis. Crystal size was calculated using the Scherrer equation. Cytotoxicity was studied in vitro using the MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) cytotoxicity assay. The optical density (OD) of each well was obtained and recorded at 570 nm for 24 h (t1), 48 h (t2), 72 h (t3), and 5 days (t4) using a microplate reader.

Results: The results of Si-Zn-doped HA showed a high specific surface area with an irregular nano-sized spherical particle structure. The atomic percentage provided the ratio of calcium to phosphate; for non-doped HA, the atomic Ca/P ratio was 1.6, but for Si-Zn-doped HA, where Zn+2 Ca and Si + replaced 4 substituted P, the atomic ratio (Ca + Zn)/(P + Si) was 1.76. The average crystal size of Si-Zn-doped HA was 46 nm, while for non-doped HA it was 61 nm. both samples were non-toxic and statistically significantly less viable than the control group After 5 days, the mean cell viability of Si-Zn-doped HA (79.17 ± 2.18) was higher than that of non-doped HA (76.26 ± 1.71) (P = 0.091).

Conclusions: The MTT assay results showed that Si-Zn-doped HA is biocompatible. In addition, it showed characteristic physiochemical properties of a large surface area with interconnected porosity.

Citing Articles

Application of Nanohydroxyapatite in Medicine-A Narrative Review.

Lubojanski A, Zakrzewski W, Samol K, Bieszczad-Czaja M, Switala M, Wiglusz R Molecules. 2024; 29(23.

PMID: 39683785 PMC: 11643452. DOI: 10.3390/molecules29235628.

References

Pon-On W, Suntornsaratoon P, Charoenphandhu N, Thongbunchoo J, Krishnamra N, Tang I . Hydroxyapatite from fish scale for potential use as bone scaffold or regenerative material. Mater Sci Eng C Mater Biol Appl. 2016; 62:183-9. DOI: 10.1016/j.msec.2016.01.051. View

Ejserholm F, Stegmayr J, Bauer P, Johansson F, Wallman L, Bengtsson M . Biocompatibility of a polymer based on Off-Stoichiometry Thiol-Enes + Epoxy (OSTE+) for neural implants. Biomater Res. 2015; 19:19. PMC: 4578262. DOI: 10.1186/s40824-015-0041-3. View

Fosca M, Streza A, Antoniac I, Vadala G, Rau J . Ion-Doped Calcium Phosphate-Based Coatings with Antibacterial Properties. J Funct Biomater. 2023; 14(5). PMC: 10219342. DOI: 10.3390/jfb14050250. View

Lebre F, Sridharan R, Sawkins M, Kelly D, OBrien F, Lavelle E . The shape and size of hydroxyapatite particles dictate inflammatory responses following implantation. Sci Rep. 2017; 7(1):2922. PMC: 5462791. DOI: 10.1038/s41598-017-03086-0. View

Muniz F, Miranda M, Dos Santos C, Sasaki J . The Scherrer equation and the dynamical theory of X-ray diffraction. Acta Crystallogr A Found Adv. 2016; 72(Pt 3):385-90. DOI: 10.1107/S205327331600365X. View

Noorani T, Luddin N, Ab Rahman I, Masudi S . In Vitro Cytotoxicity Evaluation of Novel Nano-Hydroxyapatite-Silica Incorporated Glass Ionomer Cement. J Clin Diagn Res. 2017; 11(4):ZC105-ZC109. PMC: 5449899. DOI: 10.7860/JCDR/2017/24753.9739. View

Von Euw S, Wang Y, Laurent G, Drouet C, Babonneau F, Nassif N . Bone mineral: new insights into its chemical composition. Sci Rep. 2019; 9(1):8456. PMC: 6560110. DOI: 10.1038/s41598-019-44620-6. View

Molenda M, Kolmas J . The Role of Zinc in Bone Tissue Health and Regeneration-a Review. Biol Trace Elem Res. 2023; 201(12):5640-5651. PMC: 10620276. DOI: 10.1007/s12011-023-03631-1. View

Campos J, Sousa A, Pinto P, Ribeiro J, Franca M, Caseiro A . Application of Bonelike® as synthetic bone graft in orthopaedic and oral surgery in veterinary clinical cases. Biomater Res. 2019; 22:38. PMC: 6310926. DOI: 10.1186/s40824-018-0150-x. View

10.

Mishchenko O, Yanovska A, Kosinov O, Maksymov D, Moskalenko R, Ramanavicius A . Synthetic Calcium-Phosphate Materials for Bone Grafting. Polymers (Basel). 2023; 15(18). PMC: 10536599. DOI: 10.3390/polym15183822. View

11.

Friederichs R, Chappell H, Shepherd D, Best S . Synthesis, characterization and modelling of zinc and silicate co-substituted hydroxyapatite. J R Soc Interface. 2015; 12(108):20150190. PMC: 4528584. DOI: 10.1098/rsif.2015.0190. View

12.

Lock J, Liu H . Nanomaterials enhance osteogenic differentiation of human mesenchymal stem cells similar to a short peptide of BMP-7. Int J Nanomedicine. 2011; 6:2769-77. PMC: 3218588. DOI: 10.2147/IJN.S24493. View

13.

Ferraz M . Bone Grafts in Dental Medicine: An Overview of Autografts, Allografts and Synthetic Materials. Materials (Basel). 2023; 16(11). PMC: 10254799. DOI: 10.3390/ma16114117. View

14.

Jiang S, Wang M, He J . A review of biomimetic scaffolds for bone regeneration: Toward a cell-free strategy. Bioeng Transl Med. 2021; 6(2):e10206. PMC: 8126827. DOI: 10.1002/btm2.10206. View

15.

Loncar Brzak B, Aleksijevic L, Vindis E, Kordic I, Granic M, Vidovic Juras D . Osteonecrosis of the Jaw. Dent J (Basel). 2023; 11(1). PMC: 9858620. DOI: 10.3390/dj11010023. View

16.

Swain S, Bhaskar R, Gupta M, Sharma S, Dasgupta S, Kumar A . Mechanical, Electrical, and Biological Properties of Mechanochemically Processed Hydroxyapatite Ceramics. Nanomaterials (Basel). 2021; 11(9). PMC: 8466523. DOI: 10.3390/nano11092216. View

17.

Luo J, Walker M, Xiao Y, Donnelly H, Dalby M, Salmeron-Sanchez M . The influence of nanotopography on cell behaviour through interactions with the extracellular matrix - A review. Bioact Mater. 2022; 15:145-159. PMC: 8940943. DOI: 10.1016/j.bioactmat.2021.11.024. View

18.

Hing K, Revell P, Smith N, Buckland T . Effect of silicon level on rate, quality and progression of bone healing within silicate-substituted porous hydroxyapatite scaffolds. Biomaterials. 2006; 27(29):5014-26. DOI: 10.1016/j.biomaterials.2006.05.039. View

19.

Rohmadi R, Harwijayanti W, Ubaidillah U, Triyono J, Diharjo K, Utomo P . In Vitro Degradation and Cytotoxicity of Eggshell-Based Hydroxyapatite: A Systematic Review and Meta-Analysis. Polymers (Basel). 2021; 13(19). PMC: 8512377. DOI: 10.3390/polym13193223. View

20.

Wang B, Zhang Z, Pan H . Bone Apatite Nanocrystal: Crystalline Structure, Chemical Composition, and Architecture. Biomimetics (Basel). 2023; 8(1). PMC: 10046636. DOI: 10.3390/biomimetics8010090. View