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Introduction: Mineralization of hydrogel biomaterials is considered desirable to improve their suitability as materials for bone regeneration[1],[2]. Hydrogels have been most commonly mineralized with calcium phosphate (CaP), but hydrogel-CaCO3 composites have received less attention. Magnesium (Mg) has been added to CaP to stimulate cell adhesion and proliferation and bone regeneration in vivo, but its effect as a component of carbonate-based biomaterials remains uninvestigated. In this study, gellan gum (GG) hydrogels were mineralized enzymatically with (CaCO3), Mg-enriched CaCO3 and magnesium carbonate to generate composite biomaterials for bone regeneration. GG is an inexpensive, biotechnologically produced anionic polysaccharide, from which hydrogels for cartilage regeneration have been formed by crosslinking with divalent ions[3].Methods: GG hydrogels were loaded with the enzyme urease by incubation in 5% (w/v) urease solution and mineralized for 5 days in five different media denoted as UA, UB, UC, UD and UE, which contained urea (0.17 M) and different concentrations of CaCl2 and MgCl2 (270:0, 202.5:67.5, 135:135, 67.5:202.5 and 0:250, respectively (mmol dm-3)). Discs were autoclaved and subjected to physiochemical, mechanical and cell biological characterization.Results: FTIR, SEM, TGA and XRD analysis revealed that increasing magnesium concentration decreased mineral crystallinity. At low magnesium concentrations calcite was formed, while at higher concentrations magnesian calcite was formed. Hydromagnesite formed at high magnesium concentration in the absence of calcium. Amount of mineral formed and compressive strength decreased with increasing magnesium concentration in the mineralization medium. ICP analysis revealed that Ca:Mg elemental ratio in the mineral formed was higher than in the respective mineralization media. Mineralization of hydrogels promoted adhesion and growth of osteoblast-like cells, which were supported best on mineralized hydrogels containing no or little magnesium. Hydrogels mineralized with hydromagnesite displayed higher cytotoxicity.Discussion: Enzymatic mineralization of GG hydrogels with CaCO3 in the form of calcite successfully reinforced hydrogels and promoted osteoblast-like cell adhesion and growth, but Mg enrichment had no positive effect. This is in contrast with other studies reporting that incorporation of Mg into GG mineralized with CaP promotes cell adhesion and proliferation[4]. Conclusion: Sample groups UA and UB seem to be the most promising due to the superior amount of mineral formed and cell adhesion and proliferation.

Item does not contain fulltext Hydrogels of biocompatible calcium-crosslinkable polysaccharide gellan gum (GG) were enriched with bioglass particles to enhance (i) mineralization with calcium phosphate (CaP); (ii) antibacterial properties and (iii) growth of bone-forming cells for future bone regeneration applications. Three bioglasses were compared, namely one calcium-rich and one calcium-poor preparation both produced by a sol-gel technique (hereafter referred to as A2 and S2, respectively) and one preparation of composition close to that of the commonly used 45S5 type (hereafter referred to as NBG). Incubation in SBF for 7 d, 14 d and 21 d caused apatite formation in bioglass-containing but not in bioglass-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. Mechanical testing revealed an increase in compressive modulus in samples containing S2 and NBG but not A2. Antibacterial testing using biofilm-forming meticillin-resistant staphylococcus aureus (MRSA) showed markedly higher antibacterial activity of samples containing A2 and S2 than samples containing NBG and bioglass-free samples. Cell biological characterization using rat mesenchymal stem cells (rMSCs) revealed a stimulatory effect of NBG on rMSC differentiation. The addition of bioglass thus promotes GG mineralizability and, depending on bioglass type, antibacterial properties and rMSC differentiation.

Hydrogels offer several advantages as biomaterials for bone regeneration, including ease of incorporation of soluble substances such as mineralization-promoting enzymes and antibacterial agents. Mineralization with calcium phosphate (CaP) increases bioactivity, while antibacterial activity reduces the risk of infection. Here, gellan gum (GG) hydrogels were enriched with alkaline phosphatase (ALP) and/or Seanol(®), a seaweed extract rich in phlorotannins (brown algae-derived polyphenols), to induce mineralization with CaP and increase antibacterial activity, respectively. The sample groups were unmineralized hydrogels, denoted as GG, GG/ALP, GG/Seanol and GG/Seanol/ALP, and hydrogels incubated in mineralization medium (0.1 M calcium glycerophosphate), denoted as GG/ALP_min, GG/Seanol_min and GG/Seanol/ALP_min. Seanol(®) enhanced mineralization with CaP and also increased compressive modulus. Seanol(®) and ALP interacted in a non-covalent manner. Release of Seanol(®) occurred in a burst phase and was impeded by ALP-mediated mineralization. Groups GG/Seanol and GG/ALP/Seanol exhibited antibacterial activity against methicillin-resistant Staphylococcus aureus. GG/Seanol/ALP_min, but not GG/Seanol_min, retained some antibacterial activity. Eluates taken from groups GG/ALP_min, GG/Seanol_min and GG/ALP/Seanol_min displayed comparable cytotoxicity towards MG-63 osteoblast-like cells. These results suggest that enrichment of hydrogel biomaterials with phlorotannin-rich extracts is a promising strategy to increase mineralizability and antibacterial activity.