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In Situ Hydroxyapatite Content Affects the Cell Differentiation on Porous Chitosan/Hydroxyapatite Scaffolds

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Abstract

Highly porous chitosan/hydroxyapatite composite structures with different weight ratios (100/0; 90/10; 80/20; 70/30; 60/40; 50/50; 40/60) have been prepared by precipitation method and freeze-gelation technique using calcite, urea phosphate and chitosan as starting materials. The composition of prepared composite scaffolds was characterized by X-ray diffraction analysis and Fourier transformed infrared spectroscopy, while morphology of scaffolds was imaged by scanning electron microscopy. Mercury intrusion porosimetry measurements of prepared scaffolds have shown different porosity and microstructure regarding to the HA content, along with SEM observations of scaffolds after being immersed in physiological medium. The results of swelling capacity and compressive strength measured in Dulbecco’s phosphate buffer saline (DPBS) have shown higher values for composite scaffolds with lower in situ HA content. Viability, proliferation and differentiation of MC3T3-E1 cells seeded on different scaffolds have been evaluated by live dead assay and confocal scan microscopy. Our results suggest that the increase of HA content enhance osteoblast differentiation confirming osteogenic properties of highly porous CS/HA scaffolds for tissue engineering applications in bone repair.

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Acknowledgment

The financial support of the Croatian Science Foundation (project: “Development of Biocompatible Hydroxyapatite Based Materials for Bone Tissue Engineering Applications”) and L’Oréal-UNESCO Foundation ‘For Women in Science’ is gratefully acknowledged. The financial support from the Spanish Ministry of Economy and Competitiveness and the Feder funds through the MAT2013-46467-C4-1-R project is acknowledged by the Spanish co-authors. CIBER-BBN is an initiative funded by the VI National R&D&I Plan 2008-2011, Iniciativa Ingenio 2010, Consolider Program. CIBER Actions are financed by the Instituto de Salud Carlos III with assistance from the European Regional Development Fund. The authors want to acknowledge Pilar Gómez Tena and Sergio Mestre Beltrán from Instituto de Tecnología Cerámica, Castellon, Spain, for theirs assistance with porosity measurements.

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Authors and Affiliations

  1. Faculty of Chemical Engineering and Technology, University of Zagreb, Marulićev trg 19, p.p.177, 10001, Zagreb, Croatia

    Anamarija Rogina, Marica Ivanković & Hrvoje Ivanković

  2. Center for Biomaterials and Tissue Engineering (CBIT), Universitat Politècnica de València, Camino de Vera s/n, 46022, Valencia, Spain

    Patricia Rico & Gloria Gallego Ferrer

  3. Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Zaragoza, Spain

    Patricia Rico & Gloria Gallego Ferrer

Authors
  1. Anamarija Rogina
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  2. Patricia Rico
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  3. Gloria Gallego Ferrer
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  4. Marica Ivanković
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  5. Hrvoje Ivanković
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Correspondence to Anamarija Rogina.

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Associate Editor Jane Grande-Allen oversaw the review of this article.

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Rogina, A., Rico, P., Gallego Ferrer, G. et al. In Situ Hydroxyapatite Content Affects the Cell Differentiation on Porous Chitosan/Hydroxyapatite Scaffolds. Ann Biomed Eng 44, 1107–1119 (2016). https://doi.org/10.1007/s10439-015-1418-0

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  • DOI: https://doi.org/10.1007/s10439-015-1418-0

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