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Injectable self-gelling composites for bone tissue engineering based on gellan gum hydrogel enriched with different bioactive glasses

Timothy E. Douglas1*, Wojciech Piwowarczyk2, Jana Liskova3, David Schaubroeck4*, Sander C. Leeuwenburgh5*, Gilles Brackman6*, Lieve Balcaen7*, Rainer Detsch8*, Heidi Declercq9, Katarzyna Cholewa-Kowalska10*, Agnieszka Dokupil2*, Vincent M. Cuijpers5*, Frank Vanhaecke7*, Ria Cornelissen9*, Tom Coenye6*, Aldo R. Boccaccini8* and Elżbieta Pamuła2*
  • 1 Ghent University, Department of Organic Chemistry, Belgium
  • 2 AGH University of Science and Technology, Department of Biomaterials, Poland
  • 3 Institute of Physiology, Academy of Sciences of the Czech Republic, Dept of Biomaterials and Tissue Engineering, Czechia
  • 4 Ghent University, Center for Microsystems Technology, Belgium
  • 5 Radboud University Medical Center Nijmegen, Department of Biomaterials, Netherlands
  • 6 Ghent University, Laboratory of Pharmaceutical Microbiology, Belgium
  • 7 Ghent University, Department of Analytical Chemistry, Belgium
  • 8 -Alexander Universität Erlangen-Nürnberg, Institute of Biomaterials (WW7), Germany
  • 9 Ghent University, Department of Basic Medical Science – Histology Group, Belgium
  • 10 AGH University of Science and Technology, Department of Glass Technology and Amorphous Coatings, Poland

Introduction: Gellan gum (GG) is an anionic, calcium-binding polysaccharide produced by bacteria which has been applied in hydrogel form as a tissue engineering (TE) scaffold[1]. Gelation of GG is induced by addition of divalent cations. GG’s high affinity for calcium ions means that it is highly mineralizable with calcium phosphate (CaP), which is desirable to improve the material’s bioactivity, mechanical properties and osteoblast adhesion and proliferation[2].

Here, GG hydrogels were enriched with bioactive glass particles to enhance i) mineralization with calcium phosphate (CaP); ii) antibacterial properties and iii) growth of bone-forming cells for future bone regeneration applications.

Methods: Three bioactive glasses were compared. One calcium-rich and one calcium-poor preparation (hereafter referred to as A2 and S2) were produced by a sol-gel technique[3]. One preparation of composition close to that of the commonly used 45S5 type (hereafter referred to as NBG) was produced by flame synthesis[4]. 

Pre-autoclaved 0.875% (w/v) GG solution and pre-autoclaved bioactive glass suspensions were mixed intensively and then immediately cast at room temperature. Discs of diameter 6 mm and height of 1 mm were cut out and served as samples. The final concentrations of GG and bioactive glass were 0.7% and 1% w/v, respectively. Samples were then incubated in simulated body fluid (SBF), or subjected to degradation studies, mechanical testing, Micro-CT analysis, antibacterial and cell biological testing.

Results: Incubation in SBF for 7, 14 and 21 days caused apatite formation in bioactive glass-containing but not in bioactive glass-free samples, as confirmed by FTIR, XRD, SEM, ICP-OES, and measurements of dry mass, i.e. mass attributable to polymer and mineral and not water. Dry mass increases were higher for samples containing S2 and A2 than for those containing NBG. Mechanical testing revealed an increase in compressive modulus in samples containing S2 and NBG but not A2. Samples containing A2 and S2 displayed higher antibacterial activity than samples containing NBG and bioactive glass-free samples against biofilms of meticillin-resistant Staphylococcus aureus (MRSA). Cell biological characterization using rat mesenchymal stem cells (rMSCs) revealed a stimulatory effect of NBG on rMSC differentiation.

Discussion: The greater stimulatory effect of S2 and A2 on mineralization and their higher antibacterial effect in comparison to NBG may possibly be due to higher solubility and release of calcium ions, as they are sol-gel derived, while NBG is melt-derived.

Conclusion: The addition of bioactive glass thus promotes GG mineralizability and, depending on bioactive glass type, antibacterial properties and rMSC differentiation.

T.E.L.D. acknowledges the Research Foundation Flanders (FWO) for financial support. This study was supported from National Science Centre, Poland (2012/05/B/ST8/00129)

References:
[1] Oliveira JT, et al.. J Biomed Mater Res A 2010;93(3):852-63
[2] Douglas T, et al., J Tissue Eng Regen Med 2014;8:906–918
[3] Laczka M, et al. J Biomed Mater Res 2000;52(4):601-12
[4] Brunner TJ, et al., Chem Commun (Camb) 2006(13):1384-6

Keywords: Hydrogel, Biomimetic, Scaffold, Intelligent gel

Conference: 10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016.

Presentation Type: Poster

Topic: Biomaterials in constructing tissue substitutes

Citation: Douglas TE, Piwowarczyk W, Liskova J, Schaubroeck D, Leeuwenburgh SC, Brackman G, Balcaen L, Detsch R, Declercq H, Cholewa-Kowalska K, Dokupil A, Cuijpers VM, Vanhaecke F, Cornelissen R, Coenye T, Boccaccini AR and Pamuła E (2016). Injectable self-gelling composites for bone tissue engineering based on gellan gum hydrogel enriched with different bioactive glasses. Front. Bioeng. Biotechnol. Conference Abstract: 10th World Biomaterials Congress. doi: 10.3389/conf.FBIOE.2016.01.01989

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Received: 27 Mar 2016; Published Online: 30 Mar 2016.

* Correspondence:
Dr. Timothy E Douglas, Ghent University, Department of Organic Chemistry, Ghent, Belgium, Timothy.Douglas@ugent.be
Dr. David Schaubroeck, Ghent University, Center for Microsystems Technology, Ghent, Belgium, David.Schaubroeck@elis.ugent.be
Dr. Sander C Leeuwenburgh, Radboud University Medical Center Nijmegen, Department of Biomaterials, Nijmegen, Netherlands, s.leeuwenburgh@dent.umcn.nl
Dr. Gilles Brackman, Ghent University, Laboratory of Pharmaceutical Microbiology, Ghent, Belgium, gilles.brackman@ugent.be
Dr. Lieve Balcaen, Ghent University, Department of Analytical Chemistry, Ghent, Belgium, lieve.balcaen@ugent.be
Dr. Rainer Detsch, -Alexander Universität Erlangen-Nürnberg, Institute of Biomaterials (WW7), Erlangen, Germany, rainer.detsch@ww.uni-erlangen.de
Dr. Katarzyna Cholewa-Kowalska, AGH University of Science and Technology, Department of Glass Technology and Amorphous Coatings, Kraków, Poland, cholewa@agh.edu.pl
Dr. Agnieszka Dokupil, AGH University of Science and Technology, Department of Biomaterials, Kraków, Poland, a.dokupil@gmail.com
Dr. Vincent M Cuijpers, Radboud University Medical Center Nijmegen, Department of Biomaterials, Nijmegen, Netherlands, Vincent.Cuijpers@radboudumc.nl
Dr. Frank Vanhaecke, Ghent University, Department of Analytical Chemistry, Ghent, Belgium, frank.vanhaecke@ugent.be
Dr. Ria Cornelissen, Ghent University, Department of Basic Medical Science – Histology Group, Ghent, Belgium, Ria.Cornelissen@UGent.be
Dr. Tom Coenye, Ghent University, Laboratory of Pharmaceutical Microbiology, Ghent, Belgium, Tom.Coenye@Ugent.be
Dr. Aldo R Boccaccini, -Alexander Universität Erlangen-Nürnberg, Institute of Biomaterials (WW7), Erlangen, Germany, aldo.boccaccini@ww.uni-erlangen.de
Dr. Elżbieta Pamuła, AGH University of Science and Technology, Department of Biomaterials, Kraków, Poland, Email1