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. 2016 Feb 23;51(2):224-30.
doi: 10.1016/j.rboe.2016.02.006. eCollection 2016 Mar-Apr.

Effects of ionizing radiation on proteins in lyophilized or frozen demineralized human bone

Uri Antebi  1 , Monica Beatriz Mathor  2 , André Ferreira da Silva  3 , Rodrigo Pereira Guimarães  4 , Emerson Kiyoshi Honda  4
Affiliations

Effects of ionizing radiation on proteins in lyophilized or frozen demineralized human bone

Uri Antebi et al. Rev Bras Ortop. .

Abstract
in English, Portuguese

Objective: The aim was to study the effects of application of ionizing radiation (gamma and electrons) as sterilizing agents at doses of 15 kGy, 25 kGy and 50 kGy, on lyophilized or frozen demineralized bone tissue for use in transplants.

Methods: Five human femoral diaphyses from different donors of musculoskeletal tissue were demineralized and preserved as lyophilized or frozen at -80 °C. The samples were divided into two groups: non-irradiated (control) and irradiated by means of gamma rays or an electron beam. The bone proteins were extracted and used to determine the concentrations of total protein and BMP 2 and 7.

Results: Decreases in total protein and BMP 2 and 7 concentrations were observed. The decreases in total protein concentrations, in comparison with the respective control groups, were significant in the lyophilized and frozen samples that were irradiated at a dose of 50 kGy of gamma radiation and electron beam, with reductions of more than 30%. Significant decreases in the levels of BMP 2 and 7 were also observed at higher doses and especially through use of the electron beam.

Conclusion: The reductions in the concentrations of total proteins and osteoinductive proteins (BMP 2 and 7) were related to the radiation dose, i.e. they increased with higher doses of ionizing radiation type and the type of bone preservation. The largest reductions in concentrations were observed in the bones irradiated by means of an electron beam and at a dose of 50 kGy. However, this type of radiation and this high dose are not usual practices for sterilization of bone tissue.

Objetivo: Estudar os efeitos da aplicação das radiações ionizantes (gama e elétrons) como agentes esterilizantes, nas doses de 15 kGy, 25 kGy e 50 kGy, nos tecidos ósseos desmineralizados congelados e liofilizados para uso em transplantes.

Métodos: Cinco diáfises femorais humanas de doadores distintos de tecidos musculoesqueléticos foram desmineralizadas e preservadas como liofilizadas ou congeladas a −80 °C. As amostras foram divididas em grupos não irradiados (controle) e irradiados por raios gama ou feixe de elétrons. As proteínas ósseas foram extraídas e dosadas as concentrações de proteínas totais, BMP 2 e 7.

Resultados: Foi observada diminuição das concentrações de proteínas totais e BMP 2 e 7. A diminuição das concentrações de proteínas totais, quando comparada com o respectivo controle, foi significativa nos grupos de amostras liofilizadas e congeladas e irradiadas na dose de 50 kGy por radiação gama e feixe de elétrons com redução superiores a 30%. A diminuição significativa nas concentrações das BMP 2 e 7 também foi observada nas maiores doses e principalmente por feixe de elétrons.

Conclusão: As reduções nas concentrações das proteínas totais e em proteínas osteoindutoras (BMP 2 e 7) foram relacionadas à dose de radiação, ou seja, aumentam com maiores doses, tipo de radiação ionizante e ao tipo de preservação dos ossos. As maiores reduções das concentrações foram observadas nos ossos irradiados por feixe de elétrons e na dose de 50 kGy. Porém esse tipo de radiação e essa alta dose não são práticas usuais para a esterilização dos tecidos ósseos.

Keywords: Bone and bones; Bone morphogenetic protein 2; Bone morphogenetic protein 7; Radiation, ionizing; Tissue banks.

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Figures

Fig. 1
Fig. 1
Effects of ionizing radiation on the proteins of demineralized bone tissue. Quantification of the total proteins extracted from organic bone matrix in relation to the irradiated and non-irradiated (control) groups. Representation of the results from assaying total proteins per group. (*) indicates a statistically significant difference in relation to the control (p < 0.05).
Fig. 2
Fig. 2
Effects of gamma and electron-beam radiation on the concentrations of concentrations of BMP-2 per gram of demineralized bone tissue in the groups studied. (*) indicates a statistically significant difference in relation to the control (p < 0.05).
Fig. 3
Fig. 3
Effects of gamma and electron-beam radiation on the concentrations of concentrations of BMP-7 per gram of demineralized bone tissue in the groups studied. (*) indicates a statistically significant difference in relation to the control (p < 0.05).
Fig. 4
Fig. 4
Relationship between the concentrations of BMP-2 and specific total proteins in each lyophilized tissue group: non-irradiated and irradiated with gamma and electron-beam radiation. (*) indicates a statistically significant difference in relation to the control (p < 0.05).
Fig. 5
Fig. 5
Relationship between the concentrations of BMP-2 and specific total proteins in each frozen tissue group: non-irradiated and irradiated with gamma and electron-beam radiation. (*) indicates a statistically significant difference in relation to the control (p < 0.05).
Fig. 6
Fig. 6
Relationship between the concentrations of BMP-7 and specific total proteins in each lyophilized tissue group: non-irradiated and irradiated with gamma and electron-beam radiation. (*) indicates a statistically significant difference in relation to the control (p < 0.05).
Fig. 7
Fig. 7
Relationship between the concentrations of BMP-7 and specific total proteins in each frozen tissue group: non-irradiated and irradiated with gamma and electron-beam radiation. (*) indicates a statistically significant difference in relation to the control (p < 0.05).
See this image and copyright information in PMC

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