Synthesis of Hydroxyapatite-like Ceramics from Caiman latirostris Eggshells for Biomaterial Applications
Article Sidebar
Main Article Content
Abstract
Introduction: Bone tissue engineering requires the development of biomaterials capable of reproducing the functions of the extracellular matrix and promoting bone regeneration.
Objectives: To obtain and characterize hydroxyapatite from the calcium present in Caiman latirostris eggshells, promoting an innovative and sustainable strategy for biomaterial development, and to preliminarily evaluate its biocompatibility through in vitro assays.
Materials and methods: In this study, hydroxyapatite was synthesized from Caiman latirostris eggshells obtained from the “Proyecto Yacaré” in Santa Fe Province (Argentina). The physicochemical properties of ceramics obtained with and without heat treatment (800 °C, 2 h) were evaluated, and their in vitro biological behavior was analyzed.
Results: Heat treatment produced a significant increase in the average particle size due to agglomeration, without substantially modifying the Ca/P ratio. Structural analyses indicated the formation of AB-type carbonated hydroxyapatite, with no pyrophosphate detected, which, according to the literature, is favorable for bone mineralization. Assays using MC3T3E1 osteoblastic cells showed that both ceramics are biocompatible and stimulate cell proliferation within a specific concentration range, without cytotoxic effects at higher levels.
Conclusions: The use of Caiman latirostris eggshells represents a sustainable approach aligned with the principles of green chemistry, adding value to a natural biological waste material obtained within the framework of a conservation and sustainable-use program for the species, thereby reducing environmental impact. Nevertheless, additional biocompatibility studies and in vivo assays are required to validate their safety and efficacy for clinical applications in bone tissue engineering.
Article Details
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.
La revista utiliza la licencia Creative Commons BY-NC-SA (Reconocimiento-No Comercial-Compartir Igual), que permite a los usuarios compartir y adaptar el material publicado bajo ciertas condiciones. Los autores deben ser reconocidos de acuerdo con los términos establecidos por la licencia, y los trabajos derivados solo pueden ser utilizados para fines no comerciales.
Los autores conservan el derecho de reutilizar, reproducir y difundir su trabajo en otras publicaciones o repositorios, siempre que se respeten los términos de la licencia mencionada y se cite la publicación original en la revista.
References
Marotta MV, Rojas AH, Oberti TG, Fernández JM. Obtención, caracterización y biocompatibilidad de hidrogeles poliméricos para Ingeniería de Tejidos: Estudio preliminar. Actualizaciones en Osteología 2024;20(1):9-23.
Gehrke AE, Mendoza-Bertelli A, Ledo C, et al. Neutralization of Staphylococcus aureus Protein A Prevents Exacerbated Osteoclast Activity and Bone Loss during Osteomyelitis. Antimicrob Agents Chemother2023;(67):e01140-22.
Torres ML, Cuba AH, Oberti TG, Fernández JM. Alginate-HEMA Hydrogels as Promising Biomaterials for Bone Regeneration: In Vitro and In Vivo Studies. J Biomed Mater Res B Appl Biomater 2024;112(11): e35493.
Hurtado Cuba AG, Borgeaud M, Belluzo MS, Oberti TG, Fernández JM. Estudio de Biocompatibilidad de Matrices Poliméricas Combinadas, Responsivas al pH, con Potencial Aplicación en Ingeniería de Tejido Óseo. Actual. Osteol 2023;19(2):128-43.
Selim M, Mousa HM, Abdel-Jaber GT, Barhoum A, Abdal-hay A. Innovative designs of 3D scaffolds for bone tissue regeneration: Understanding principles and addressing challenges. European Polymer Journal 2024;215:113251.
Fernández JM, Molinuevo MS, Cortizo MS, Cortizo AM. Development of an osteoconductive PCL–PDIPF–hydroxyapatite composite scaffold for bone tissue engineering. J Tissue Eng Regen Med 2011;5: e126-e135.
Bauso LV, La Fauci V, Longo C, Calabrese G. Bone Tissue Engineering and Nanotechnology: A Promising Combination for Bone Regeneration. Biology 2024;13:237.
Fernández JM, Cortizo MS, Cortizo AM, Abraham GA. Osteoblast Behavior on Novel Porous Polymeric Scaffolds. J. Biomater. Tissue Eng 2011;1:86-92.
Garimella A, Ghosh SB, Bandyopadhyay-Ghosh S. Biomaterials for bone tissue engineering: achievements to date and future directions. Biomedical Material2025;20:012001.
Yadav MK, Shukla RH, Prashanth KG. A comprehensive review on development of waste derived hydroxyapatite (HAp) for tissue engineering application. Materials Today: Proceedings 2023.
Gnanasekaran R, Yuvaraj D, Mathan Muthu DM, et al. Extraction and characterization of biocompatible hydroxyapatite (Hap) from red big eye fish bone: Potential for biomedical applications and reducing biowastes. Sustainable Chemistry for the Environment 2024;7:100142,
Zamri MFMA, Bahru R, Amin R, et al. Waste to health: A review of waste derived materials for tissue engineering. Journal of Cleaner Production 2021; 290: 125792.
Cedillo-Leal C, Simoncini MS, Leiva PML, Larriera A, Lang JW, Piña CI. Eggshell structure in Caiman latirostris eggs improves embryo survival during nest inundation. Proc R Soc B 2017; 284: 20162675.
Fernández MS, Simoncini MS, Dyke G. Irregularly calcified eggs and eggshells of Caiman latirostris (Alligatoridae: Crocodylia). Naturwissenschaften2013;100(5):451-7.
Piazza MV, Fernández MS, Leiva PML, Piña CI, Simoncini MS. “Intracascaral space” an eggshell structure of Caiman latirostris eggs. Sci Rep 2021;11:5579.
Castro MAM, Portela TO, Correa GS, et al.. Synthesis of hydroxyapatite by hydrothermal and microwave irradiation methods from biogenic calcium source varying pH and synthesis time. Boletín de la Sociedad Española de Cerámica y Vidrio2022; 61:35-41.
Kamalanathan P, Ramesh S, Bang LT, et al. Synthesis and sintering of hydroxyapatite derived from eggshells as a calcium precursor. Ceramics International 2014;40(10):16349-59.
Fernández JM, Molinuevo MS, Sedlinsky C, Schurman L, Cortizo AM, McCarthy AD. Strontium ranelate prevents the deleterious action of advanced glycation endproducts on osteoblastic cells via calcium channel activation. Eur J Pharmacol 2013; 706(1-3):41-7.
Larriera A, Imhof A. En: Bolkovic, M L, Ramadori D (eds.). Manejo de Fauna Silvestre en la Argentina. Programas de uso sustentable. Buenos Aires: Dirección de Fauna Silvestre, Secretaría de Ambiente y Desarrollo Sustentable; 2006. pp. 51-64.
Mostafa NY. Characterization, thermal stability and sintering of hydroxyapatite powders prepared by different routes. Materials Chemistry and Physics 2005;94:333-41.
Amin AMM, Besisa DHA, El-Amir AAM, Zaki ZI, Ahmed YMZ. Role of heat treatment of hydroxyapatite powder prior to suspension preparation on the suspension flow behavior. Open Ceramics 2022;9:100239.
Klecandova L, Sliva A, Šindler M, et al. Effect of Processing on Structural and Morphological Changes of Hydroxyapatite-Based Materials. Chemistry Select 2025;10: e202405109.
Diichuk V, Diichuk I. Thermal treatment effect on the particle size distribution of alkaline earth metals hydroxyapatite. Scientiae Radices 2023; 2(4):309-18.
Addison WN, Azari F, Sørensen ES, Kaartinen MT, McKee MD. Pyrophosphate Inhibits Mineralization of Osteoblast Cultures by Binding to Mineral, Up-regulating Osteopontin, and Inhibiting Alkaline Phosphatase Activity. J Biol Chem. 2007;282(21):15872-83.
Jaber H, Mousa MH, Qadir A, et al. Comprehensive investigation into thermal stability of AB-type bio- carbonate hydroxyapatite synthesized via heat-treated bovine bone. Open Ceramics 2024;20:100701.
Maeno S, Niki Y, Matsumoto H, et al. The effect of calcium ion concentration on osteoblast viability, proliferation and differentiation in monolayer and 3D culture. Biomaterials 2005;26(23): 4847-55.
Pieters IY, Van den Vreken NMF, Declercq HA, Cornelissen MJ, Verbeeck RMH. Carbonated apatites obtained by the hydrolysis of monetite: Influence of carbonate content on adhesion and proliferation of MC3T3-E1 osteoblastic cells. Acta Biomaterialia 2010;6:1561-8.
Tsang EJ, Arakawa CK, Zuk PA, Wu BM. Osteoblast Interactions within a Biomimetic Apatite Microenvironment. Ann Biomed Eng 2011; 39(4):1186-200.
Nagai H, Kobayashi-Fujioka M, Fujisawa K, et al. Effects of low crystalline carbonate apatite on proliferation and osteoblastic differentiation of human bone marrow cells. J Mater Sci: Mater Med 2015; 26:99.
Chou YF, Huang W, Dunn JCY, Miller TA, Wu BM. The effect of biomimetic apatite structure on osteoblast viability, proliferation, and gene expression. Biomaterials 2005;26:285-95.
Poinern GJE, Brundavanam R, Le XT, Djordjevic S, Prokic M, Fawcett D. Thermal and ultrasonic influence in the formation of nanometer scale hydroxyapatite bio-ceramic. Int J Nanomedicine 2011;6, 2083-95.
Jamil M, Elouahli A, Abida F, et al. Development of Triphasic Hydroxyapatite/(α and β)‑Tricalcium Phosphate Based Composites by Sintering Powder of Calcium‑Apatite in the Presence of Montmorillonite. Journal of Inorganic and Organometallic Polymers and Materials 2020; 30(6):2489-98.
Ruiz J, Moreno D, Copete H, Vargas F, López ME. Calcium phosphate cements improved by additionof carbonated Hydroxyapatite type B. Boletín de la Sociedad Española de Cerámica y Vidrio 2023; 62(4):315-28.
Sabu U, Logesh G, Rashad M, Joy A, Balasubramanian M. Microwave assisted synthesis of biomorphic hydroxyapatite. Ceramic International 2019; 45(6):6718-22.
Ramesh S, Natasha AN, Tan CY, et al. Direct conversion of eggshell to hydroxyapatite ceramic by a sintering method. Ceramic International 2016;42(6): 7824-9.
Gheisari H, Karamian E, Abdellahi M. A novel hydroxyapatite –Hardystonite nanocomposite ceramic. Ceramics International 2015;41:5967-75.
Kesmez O. Preparation of Anti-bacterial Biocomposite Nanofibers Fabricated by Electrospinning Method. Journal of the Turkish Chemical Society Chemistry 2020;7(1):125-42.