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The Effect of Boron-Containing Nano-Hydroxyapatite on Bone Cells

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Abstract

Metabolic diseases or injuries damage bone structure and self-renewal capacity. Trace elements and hydroxyapatite crystals are important in the development of biomaterials to support the renewal of bone extracellular matrix. In this study, it was assumed that the boron-loaded nanometer-sized hydroxyapatite composite supports the construction of extracellular matrix by controlled boron release in order to prevent its toxic effect. In this context, boron release from nanometer-sized hydroxyapatite was calculated by ICP-MS as in large proportion within 1 h and continuing release was provided at a constant low dose. The effect of the boron-containing nanometer-sized hydroxyapatite composite on the proliferation of SaOS-2 osteoblasts and human bone marrow-derived mesenchymal stem cells was evaluated by WST-1 and compared with the effects of nano-hydroxyapatite and boric acid. Boron increased proliferation of mesenchymal stem cells at high doses and exhibited different effects on osteoblastic cell proliferation. Boron-containing nano-hydroxyapatite composites increased osteogenic differentiation of mesenchymal stem cells by increasing alkaline phosphatase activity, when compared to nano-hydroxyapatite composite and boric acid. The molecular mechanism of effective dose of boron-containing hydroxyapatite has been assessed by transcriptomic analysis and shown to affect genes involved in Wnt, TGF-β, and response to stress signaling pathways when compared to nano-hydroxyapatite composite and boric acid. Finally, a safe osteoconductive dose range of boron-containing nano-hydroxyapatite composites for local repair of bone injuries and the molecular effect profile in the effective dose should be determined by further studies to validation of the regenerative therapeutic effect window.

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Acknowledgments

Statistical analyses were done by Osman Dağ, MSc, research assistant at the Biostatistics Department Faculty of Medicine in Hacettepe University.

Funding

This study was supported by Hacettepe University Scientific Research Project Coordination Unit with project number TBB-2017-13-312.

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Authors and Affiliations

  1. Graduate School of Science and Engineering, Department of Bioengineering, Hacettepe University, Ankara, Turkey

    Merve Gizer

  2. Faculty of Medicine, Department of Medical Biology, Atilim University, Ankara, Turkey

    Sevil Köse

  3. Department of Medical Genetics, Hacettepe University Faculty of Medicine, Ankara, Turkey

    Beren Karaosmanoglu & Ekim Z. Taskiran

  4. Department of Analytical Chemistry, Gazi University Faculty of Pharmacy, Ankara, Turkey

    Aysel Berkkan

  5. Department of Metallurgical and Materials Engineering, Middle East Technical University Faculty of Engineering, Ankara, Turkey

    Muharrem Timuçin

  6. Department of Sports Medicine, Hacettepe University Faculty of Medicine, Ankara, Turkey

    Feza Korkusuz

  7. Department of Histology and Embryology, Hacettepe University Faculty of Medicine, 06100 Sihhiye, Ankara, Turkey

    Petek Korkusuz

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  1. Merve Gizer
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Sup. 1

The mean-standard deviation graphics of the SaOS-2 cells and hBMCs proliferation by days. Optical density (OD) values of SaOS-2 cells proliferation a on day 1, b on day 3, c on day 5 (n = 14 for B-nHAp, nHAp composites and BA treated cells and n = 16 for untreated cells at each time point) and hBMSCs proliferation d on day 1, e on day 3 and f on day 5 are shown (n = 21 for B-nHAp, nHAp composites and BA treated cells and n = 24 for untreated cells at each time point). (a, b, c) p < 0.05, only the statistically significance of the relevant data is shown. (a) refers to the statistically significant difference comparing to untreated control, (b) and (c) refer to the statistically significant difference comparing to equivalent dose of nHAp and BA within the composite respectively. hBMSCs: Human bone marrow mesenchymal stem cells, B-nHAp: Boron containing nano-hydroxyapatite, nHAp: Nano-hydroxyapatite, BA: Boric acid. (PNG 336 kb)

High resolution image (TIF 528 kb)

Sup. 2

The mean-standard deviation graphics of the hBMSCs ALP activity by days. Optical density (OD) values of the ALP activity assay a on day 7, b on day 14 and c 21 are shown (n = 6 for all groups at each time point). (a, b, c) p < 0.05, Only the statistically significance of the relevant data is shown. (a) refers to the statistically significant difference comparing to untreated control, (b) and (c) refer to the statistically significant difference comparing to equivalent dose of nHAp and BA within the composite respectively. hBMSCs: Human bone marrow mesenchymal stem cells, ALP: Alkaline phosphatase, B-nHAp: Boron containing nano-hydroxyapatite, nHAp: Nano-hydroxyapatite, BA: Boric acid. (PNG 685 kb)

High resolution image (TIF 4741 kb)

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Gizer, M., Köse, S., Karaosmanoglu, B. et al. The Effect of Boron-Containing Nano-Hydroxyapatite on Bone Cells. Biol Trace Elem Res 193, 364–376 (2020). https://doi.org/10.1007/s12011-019-01710-w

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