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Generation of composites for bone tissue-engineering applications consisting of gellan gum hydrogels mineralized with calcium and magnesium phosphate phases by enzymatic means

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Published
  • Timothy Edward Lim Douglas
  • Grzegorz Krawczyk
  • Elzbieta Pamula
  • Heidi Declercq
  • David Schaubroeck
  • Miroslaw Bucko
  • Lieve Balcaen
  • Pascal Van der Voort
  • Vitaliy Bliznuk
  • Natasja van den Vreken
  • Mamoni Dash
  • Rainer Detsch
  • Aldo Boccaccini
  • Frank Vanhaecke
  • Ria Cornelissen
  • Dubruel
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<mark>Journal publication date</mark>11/2016
<mark>Journal</mark>Journal of Tissue Engineering and Regenerative Medicine
Issue number11
Volume10
Number of pages17
Pages (from-to)938-954
Publication StatusPublished
Early online date21/02/14
<mark>Original language</mark>English

Abstract

Mineralization of hydrogels, desirable for bone regeneration applications, may be achieved enzymatically by incorporation of alkaline phosphatase (ALP). ALP‐loaded gellan gum (GG) hydrogels were mineralized by incubation in mineralization media containing calcium and/or magnesium glycerophosphate (CaGP, MgGP). Mineralization media with CaGP:MgGP concentrations 0.1:0, 0.075:0.025, 0.05:0.05, 0.025:0.075 and 0:0.1 (all values mol/dm3, denoted A, B, C, D and E, respectively) were compared. Mineral formation was confirmed by IR and Raman, SEM, ICP‐OES, XRD, TEM, SAED, TGA and increases in the the mass fraction of the hydrogel not consisting of water. Ca was incorporated into mineral to a greater extent than Mg in samples mineralized in media A–D. Mg content and amorphicity of mineral formed increased in the order A < B < C < D. Mineral formed in media A and B was calcium‐deficient hydroxyapatite (CDHA). Mineral formed in medium C was a combination of CDHA and an amorphous phase. Mineral formed in medium D was an amorphous phase. Mineral formed in medium E was a combination of crystalline and amorphous MgP. Young's moduli and storage moduli decreased in dependence of mineralization medium in the order A > B > C > D, but were significantly higher for samples mineralized in medium E. The attachment and vitality of osteoblastic MC3T3‐E1 cells were higher on samples mineralized in media B–E (containing Mg) than in those mineralized in medium A (not containing Mg). All samples underwent degradation and supported the adhesion of RAW 264.7 monocytic cells, and samples mineralized in media A and B supported osteoclast‐like cell formation.