Preparation of Hydroxyapatite Scaffolds from Cockle Shell for Bone repair

This study aimed to prepare hydroxyapatite (HA) scaffolds using a solid-state reaction with raw materials of CaCO3 and NH4H2PO4.

Authors

  • Sittiporn Punyanitya Innovative Biomaterials and Medical Device Research Group, Mae Fah Luang University, Chiang Rai, Thailand 57100
  • Rungsarit Koonawoot Boonyapanit Co., Ltd., 19/44, Singharaj Road, Tumbol Sriphum, Amphur Muang Chiang Mai, Thailand 50200
  • Anucha Raksanti Multidisciplinary Research Institute, Chiang Mai University, Chiang Mai, Thailand 50200
  • Phuwadit Thungkaew Faculty of Management Science, Sakon Nakhon Rajabhat University, Sakon Nakhon, Thailand 41000

Keywords:

Bone graft, Cockle shells, Scaffolds, Soft tissue, Histological

Abstract

Background: There is a growing demand for effective bone repair materials to replace and reduce the import of materials from abroad, utilizing waste materials from natural resources in Thailand. The goal is to create medical devices and develop inventions into innovations that can be commercialized.

Objectives: This study aimed to prepare hydroxyapatite (HA) scaffolds using a solid-state reaction with raw materials of CaCO3 and NH4H2PO4.

Materials and Method: Scaffolds were synthesized under two conditions with sintering temperatures ranging from 1100 to 1300°C for 2 hours. The phase contents, physical, and mechanical properties were investigated. Additionally, to evaluate the maximum HA phase content, each condition was assessed in the subcutaneous soft tissue of laboratory rats over 7, 30, and 90 days. Characterization techniques included XRD, SEM, porosity analysis, bending strength testing, hardness measurement, and histological studies using optical microscopy.

Results: The findings indicated that the scaffolds sintered at 1300°C for 2 hours (Condition 2) had the highest HA content. After 90 days, the scaffold’s reaction with soft tissues showed mild inflammation and good tissue compatibility due to the high HA content.

Conclusion: Our results conclude that HA scaffolds prepared from cockle shells have potential for use as bone grafts, as the samples were found to be nontoxic and biocompatible with soft tissues.

References

Palmer LC, Newcomb CJ, Kaltz SR, Spoerke ED, and Stupp SI, Biomimetic Systems for Hydroxyapatite Mineralization Inspired by Bone and Enamel. Chem Rev. 2008; 108 (11): 4754-83. https://doi.org/10.1021/cr8004422

ASTM F1185-03. Standard Specification for Composition of Hydroxyapatite for Surgical Implants, West Conshohocken, PA, USA. 2009, pp. 475-477.

Murugan R and Ramakrishna S. Development of nanocomposites for bone grafting. Comp Sci Tech. 2005: 65 (15-16): 2385-406. https://doi.org/10.1016/j.compscitech.2005.07.022

Vecchio KS, Zhang X, Massie JB, Wang M, and Kim CW. Conversion of bulk seashells to biocompatible hydroxyapatite for bone implants. Acta Biomater. 2007; 3 (6): 910-8. https://doi.org/10.1016/j.actbio.2007.06.003

Raynaud S, Champion E, Bernahe- Assollant D, and Thomas P. Calcium phosphate apatite with variable Ca/P atomic ratio I. Synthesis, characterization and thermal stability of powders. Biomater. 2002; 23 (4): 1065-72. https://doi.org/10.1016/S0142-9612(01)00218-6

Balamurugan A, Kannan S, and Rajeswari S. Bioactive sol-gel hydroxyapatite surface for biomedical application-in vitro study. Trends in Biomaterials and Artificial Organs. 2002; 16 (1): 18-20.

Zhang X, and Vecchio KS. Creation of dense hydroxyapatite (synthetic bone) by hydrothermal conversion of seashells. Materials Science and Engineering: C. 2006; 26: 1445-50. https://doi.org/10.1016/j.msec.2005.08.007

Rodriquez-Lorenzo LM, Vallet-Regi M, and Ferreira JMF. Fabrication of hydroxyapatite bodies by uniaxial pressing from a precipitated powder. Biomaterials. 2001; 22 (6): 583-8. https://doi.org/10.1016/S0142-9612(00)00218-0

Rao RR, Roopa HN and Kannan TS. Solid state synthesis and thermal stability of HAP and HAP-β-TCP composite ceramic powders. J Mater Sci: Materials in Medicine. 1997; 8 (8): 511-8. https://doi.org/10.1023/A:1018586412270

Cox SC, Mallick KK and Walton RI. Comparison of techniques for the synthesis of hydroxyapatite, Bioinspired, Biomimetic and Nanobiomaterials. 2014; 4 (1): 37-47. https://doi.org/10.1680/bbn.14.00010

Pramanik S, Agarwal AK, Rai KN, Garg A. Development of high strength hydroxyapatite by solid-state-sintering process. Ceramic International. 2007; 33, (3): 419-26. https://doi.org/10.1016/j.ceramint.2005.10.025

Kaplan DL. Mollusc shell structures: Novel design strategies for synthetic materials. Current Opinion in Solid State and Materials Science. 1998; 3: 232-6. https://doi.org/10.1016/S1359-0286(98)80096-X

Koonawoot R, Thiansem S, and Punyanitya S. Preparation and Characterization of Hydroxyapatite Powder from Mollusc Shells. Pure and Applied Chemistry International Conference (PACCON 2012), pp. 190-193.

Committee for the Update of the Guide for the Care and Use of Laboratory Animals, "Guide for the Care and Use of Laboratory Animals, 8th Ed. The National Academy of Sciences, Washington, DC, 2011.

5. Department of Health Education and Welfare, The Guide for Care and Use of Laboratory Animals, Publication (No.78-23). Av a i l a b l e from Superintendent of Documents, U.S. Government Printing Office, Washington, 20402. 1978.

ASTM F981-04. Standard Practice for Assessment of Compatibility of Biomaterials for Surgical Implants with Respect to Effect of Materials on Muscle and Bone. 2010.

ASTM F763-04, Standard Practice for Short-Term Screening of Implant Materials. 2010.

Callut S, and Knowles JC. Correlation between structure and compression strength in reticulated glass-reinforced hydroxyapatite foam, Journal of Material Science: Material in Medicine. 2000; 13 (5): 485-9. https://doi.org/10.1023/A:1014718722710

ASTM E 112-10. Standard Test Method for Determining Average Grain Size. 2010.

De With G, and Wagemans HHM. Ball-on-ring test revisited. Journal of the American Ceramic Society. 1989; 72 (8): 1538-41. https://doi.org/10.1111/j.1151-2916.1989.tb07702.x 21. Callister WD. Fundamentals of Material Science and Engineering. Willey, New York.2001.

JCPDS-ICDD Card no.9-432. International center for diffraction data, Newton Square, PA. 2000.

Raynaud S, Champion E, Bernache- Assollant D and Laval JP. Determination of Calcium/Phosphorus Atomic Ratio of Calcium Phosphate Apatites Using X-ray Diffractometry. Journal of the American Ceramic Society. 2001; 84 (2): 359-66. https://doi.org/10.1111/j.1151-2916.2001.tb00663.x

How YM, Kasim SR, Akil HM and Ahmad ZA. Effect of CaCO3 Particle Size in Synthesis of β-TCP Powder. Journal of Nuclear and related Technologies. 2009; 6 (1): 19-24.

Farzadi A, Solati-Hashjin M, Bakhshi F and Aminian A. Synthesis and characterization of hydroxyapatite/ β-tricalcium phosphate nanocomposite using microwave irradiation. Ceramics International. 2011; 37 (1): 65-71. https://doi.org/10.1016/j.ceramint.2010.08.021

Rahaman MN. Ceramic Processing And Sintering, 2nd Ed., Marcel Dekker, New York. 2003, pp. 754-755.

Yang X and Wang Z. Synthesis of biphasic ceramics of hydroxyapatite and β-tricalcium phosphate with controlled phase content and porosity. Journal of Materials Chemistry. 1998; 8 (10): 2233-7. https://doi.org/10.1039/a802067a

Guo D, Xu K and Han Y. Influence of cooling modes on purity of solid-state synthesized tetracalcium phosphate. Materials Science and Engineering: B. 2005; 116 (2): 2, 175-81. https://doi.org/10.1016/j.mseb.2004.09.032

De Bruijn JD, Sleijster M, Doel MVD, Apeldoorn AV, d Brink IV, Blitterswijk CV, et al. Porous hydroxyapatite scaffold coated with in vitro formed bone-like matrix induct faster bone formation as compared to cell seeded scaffold. Poster of the 47th annual meeting, Feb. 25-28, Ortho. Res. Soc., San Francisco, California. 2001.

Wang M. Bioactive materials and processing. Berlin, Heidelberg, Springer Verlag. 2004: pp. 1-82. 31. Jarcho M. Calcium phosphate ceramics as hard tissue prosthetics. Clinical Orthopaedics and Related Research. 1981; 157: 259-78. https://doi.org/10.1097/00003086-198106000-00037

Downloads

Published

2025-04-28

How to Cite

1.
Punyanitya S, Koonawoot R, Raksanti A, Thungkaew P. Preparation of Hydroxyapatite Scaffolds from Cockle Shell for Bone repair: This study aimed to prepare hydroxyapatite (HA) scaffolds using a solid-state reaction with raw materials of CaCO3 and NH4H2PO4. GMSMJ [internet]. 2025 Apr. 28 [cited 2025 Jun. 17];5(2):85-99. available from: https://he02.tci-thaijo.org/index.php/gmsmj/article/view/272213

Issue

Section

Innovation in Medicine