Investigating the Effects of Gamma Irradiation on Osteogenic Properties of OsvehOss Synthetic Bone Graft Substitutes as Orthopedic Implant Materials

Document Type : Research Articles

Authors

• Heydar Khadivi Ayask ¹
• Nasrin Sasani ¹
• Halimeh Hassanzadeh ^{2, 3}
• Masoud Golestanipour ⁴
• Ahmad Moloodi ⁴
• Vahide Sadat Ebrahimi ¹
• Maryam M. Matin ^{3, 5}

¹ Osveh Asia Medical Instrument Company, Mashhad, Iran

² Stem Cells and Regenerative Medicine Research Group, Academic Center for Education, Culture and Research (ACECR)-Khorasan Razavi, Mashhad, Iran

³ Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran

⁴ Materials Research Group, Academic Center for Education, Culture and Research (ACECR), Mashhad Branch, Mashhad, Iran

⁵ Novel Diagnostics and Therapeutics Research Group, Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran

https://doi.org/10.22067/jcmr.2024.87812.1094

Abstract

Bone graft substitutes are used in bone tissue engineering, orthopedics, and dentistry to help bone repair. The sterility and pyrogenicity of the bone grafts before clinical use are considered part of the regulatory requirements, however sterilization of biomaterials is challenging due to the physicochemical changes resulting from the localized increase in gamma dose during irradiation. The effects of gamma radiation dose on the biological behavior of synthetic bone grafts have not been extensively investigated. The present study aimed to evaluate the effects of gamma radiation sterilization doses on OsvehOss synthetic bone grafts via chemical, mechanical, and in vitro biological examinations. XRD analysis and compression test were carried out to evaluate the chemical and mechanical changes of synthetic bone grafts induced by the highest gamma radiation dose applied in this study. Human osteosarcoma MG-63 cells were used to assay their osteogenic response while grown on a biphasic bone graft substitute. Cell attachment and proliferation were confirmed via scanning electron microscopy on days 3, 7, and 14 of culture. Alkaline phosphatase (ALP) activity was determined to assess osteogenesis. Alizarin red S (ARS) staining was also used to identify calcium deposition in osteocytes developed after differentiation of MG-63 cells. Our results illustrated that gamma irradiation did not cause dose-dependent changes in chemical and mechanical properties of OsvehOss BCP (Biphasic Calcium Phosphate) bone grafts when the doses increased up to 50 kGy. Furthermore, OsvehOss BCP samples demonstrated high osteoconductivity in all irradiation treatment groups. ALP and ARS analyses also indicated that the application of irradiation doses up to 50 kGy for sterilization of OsvehOss BCP grafts had no significant effects on osteogenesis and calcium deposition in osteoblast cells cultured on grafts. In conclusion, OsvehOss biomaterials can be sterilized safely for biomedical applications.

Keywords

• biphasic bone graft substitutes
• gamma irradiation
• MG-63 cells
• osteogenesis

References

Arinzeh T. L., Tran T., McAlary J., Daculsi G., (2005) A comparative study of biphasic calcium phosphate ceramics for human mesenchymal stem-cell-induced bone formation. Biomaterials 26: 3631-3638.

Betz R. R., (2002) Limitations of autograft and allograft: New synthetic solutions. Orthopaedics 25:561-570.

Costa, T., Serro, A.P., Pires, E., Colaço, R., (2006) Influence of sterilization with γ-Irradiation in the degradation of plasma sprayed hydroxyapatite coatings. Materials Science Forum 514-516(2): 1054-1058.

Currey J. D., Foreman J., Laketić I., Mitchell J., Pegg D. E., Reilly G. C., (1997) Effects of ionizing radiation on the mechanical properties of human bone. Journal of Orthopaedic Research 15:111-117.

Deligianni D. D., Katsala N. D., Koutsoukos P. G., Missirlis Y. F., (2001) Effect of surface roughness of hydroxyapatite on human bone marrow cell adhesion, proliferation, differentiation and detachment strength. Biomaterials 22:87-96.

Deng Y., Liu X., Xu A., Wang L., Luo Z., Zheng Y., Deng F., Wei J., Tang Z., Wei S., (2015) Effect of surface roughness on osteogenesis in vitro and osseointegration in vivo of carbon fiber-reinforced polyetheretherketone-nanohydroxyapatite composite. International Journal of Nanomedicine 10:1425-1447.

Godette G. A., Kopta J. A., Egle D. M., (1996) Biomechanical effects of gamma irradiation on fresh frozen allografts in vivo. Orthopedics 19:649-653.

Golub E. E., Harrison G., Taylor A. G., Camper S., Shapiro I. M., (1992) The role of alkaline phosphatase in cartilage mineralization. Bone and Mineral 17:173-278.

Hannink G., Arts J. J. C., (2011) Bioresorbability, porosity and mechanical strength of bone substitutes: What is optimal for bone regeneration? Injury 42:S22-S25.

Hing K. A., Bioceramic bone graft substitutes: influence of porosity and chemistry. International Journal of Applied Ceramic Technology 2:184-199.

Kawasaki Y., Sotome S., Yoshii T., Torigoe I., Maehara H., Sugata Y., Hirano M., Mochizuki N., Shinomiya K., Okawa A., (2010) Effects of gamma ray irradiation on mechanical properties, osteoconductivity, and absorption of porous hydroxyapatite/collagen. Journal of Biomedical Materials Research B: Applied Biomaterials 92: 161-167.

Kouhestani F., Dehabadi F., Hasan Shahriari M., Motamedian S. R., (2018) Allogenic vs. synthetic granules for bone tissue engineering: an in vitro study. Progress in Biomaterials 7:133-141.

Kuboki Y., Jin Q., Kikuchi M., Mamood J., Takita H., (2002) Geometry of artificial ECM: sizes of pores controlling phenotype expression in BMP-induced osteogenesis and chondrogenesis. Connective Tissue Research 43:529-534.

Kubisz L., Mielcarek S., Jaroszyk F., (2003) Changes in thermal and electrical properties of bone as a result of 1 MGy-dose γ-irradiation. International Journal of Biological Macromolecules 33:89-93.

Liu W., Wang T., Yang C., Darvell B.W., Wu J., Lin K., Chang J., Pan H., Lu W.W., (2016) Alkaline biodegradable implants for osteoporotic bone defects—Importance of microenvironment pH. Osteoporosis International 27:93-104.

Liu W., Dan X., Lu W.W., Zhao X., Ruan C., Wang T., Cui X., Zhai X., Ma Y., Wang D., et al., (2019) Spatial distribution of biomaterial microenvironment pH and its modulatory effect on osteoclasts at the early stage of bone defect regeneration. ACS Applied Materials and Interfaces 11:9557-9572.

Malik M., Puleo D., Bizios R., Doremus R., (1992) Osteoblasts on hydroxyapatite, alumina and bone surfaces in vitro; morphology during the first 2 h of attachment. Biomaterials 13:123-128.

Nguyen H., Morgan D. A. F., Forwood M. R., (2007) Sterilization of allograft bone: effects of gamma irradiation on allograft biology and biomechanics. Cell Tissue Banking 8:93-105.

Nieminen T., Kallela I., Keranen J., Hiidenheimo I., Kainulainen H., Wuolijoki E., Rantala I., (2006) In vivo and in vitro degradation of a novel bioactive guided tissue regeneration membrane. International Journal of Oral and Maxillofacial Surgery 35:727-732.

Rohin S., Arunkumar P., (2023) Effect of sterilization techniques on biomaterial inks’ properties and 3D bioprinting parameters, Bioprinting 33:e00294.

Shahabi S., Naja F., Majdabadi A., Hooshmand T., Haghbin Nazarpak M., Karimi B. and Fatemi S. M., (2014) Effect of gamma irradiation on structural and biological properties of a PLGA-PEG-hydroxyapatite composite. The Scientific World Journal 420616.

Suwanprateeb J., Tanner K. E., S. Turner, Bonfield W., (1998) Influence of sterilization by γ-irradiation and of thermal annealling on creep of hydroxyapatite reinforced polyethylene composites. Journal of Biomedical Materials Research 39:16-22.

Takehisa, M., et al., (1998) Radiation resistance of the bioburden from medical
devices, Radiation Physics and Chemistry 52:21-27.

Triantafyllou E., Sotiropoulos E., Triantafyllou J. N., (1975) The mechanical properties of the lymphylized and Irradiated bone grafts. Acta Orthopeadica Belgica 41:35-44.

Türker N.S., Özer A.Y., Kutlu B., Nohutcu R., Sungur A., Bilgili H., Ekizoglu M., Özalp M., (2014) The effect of gamma radiation sterilization on dental biomaterials. Tissue Engineering and Regenerative Medicine 11:341-349.