The effect of inorganic additives to calcium phosphate on in vitro behavior of osteoblasts and osteoclasts

Liang Yang, Soledad Perez-Amodio, F. Barrère, Vincent Everts, Clemens van Blitterswijk, Pamela Habibovic

Research output: Contribution to journalArticleAcademicpeer-review

140 Citations (Scopus)


This study describes a medium-throughput system based on deposition of calcium phosphate films in multi-well tissue culture plates that can be used to study the effect of inorganic additives on the behavior of osteoblasts and osteoclasts in a standardized manner. All tested elements, copper, zinc, strontium, fluoride and carbonate were homogenously deposited into calcium phosphate films in varying concentrations by using a biomimetic approach. The additives affected morphology and composition of calcium phosphate films to different extent, depending on the concentration used. The effect on proliferation and differentiation of MC3T3-E1 osteoblasts depended on the compound and concentration tested. In general, copper and zinc ions showed an inhibitory effect on osteoblast proliferation, the effect of strontium was concentration dependent, whereas films containing fluoride and carbonate, respectively, augmented osteoblast proliferation. Copper and zinc had no effect or were mild inhibitory on osteoblast differentiation, while strontium, fluoride and carbonate ions demonstrated a clear decrease in differentiation in comparison to the control films without additives. Primary osteoclasts cultured on calcium phosphate films containing additives showed a significantly decreased resorptive activity as compared to the control, independent on the element incorporated. No cytotoxic effect of the elements in the concentrations tested was observed. The system presented in this study mimics bone mineral containing trace elements, making it useful for studying fundamental processes of bone formation and turnover. The present results can be used for modifying bone graft substitutes by addition of inorganic additives in order to affect their performance in bone repair and regeneration.
Original languageEnglish
Pages (from-to)2976-2989
Issue number11
Publication statusPublished - 2010


  • Osteoblast
  • Osteoclast
  • METIS-273560
  • Bone graft substitute
  • Inorganic additives
  • Medium-throughput study
  • Calcium phosphate
  • IR-80799

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