• Home
  • Search Results
  • Elucidating the individual effects of calcium and phosphate ions on hMSCs by using composite materials.

Elucidating the individual effects of calcium and phosphate ions on hMSCs by using composite materials.

Acta biomaterialia (2015-02-14)
Charlène B S S Danoux, David C Bassett, Ziryan Othman, Ana I Rodrigues, Rui L Reis, Jake E Barralet, Clemens A van Blitterswijk, Pamela Habibovic
ABSTRACT

The biological performance of bone graft substitutes based on calcium phosphate bioceramics is dependent on a number of properties including chemical composition, porosity and surface micro- and nanoscale structure. However, in contemporary bioceramics these properties are interlinked, therefore making it difficult to investigate the individual effects of each property on cell behavior. In this study we have attempted to investigate the effects of calcium and inorganic phosphate ions independent from one another by preparing composite materials with polylactic acid (PLA) as a polymeric matrix and calcium carbonate or sodium phosphate salts as fillers. Clinically relevant bone marrow derived human mesenchymal stromal cells (hMSCs) were cultured on these composites and proliferation, osteogenic differentiation and ECM mineralization were investigated with time and were compared to plain PLA control particles. In parallel, cells were also cultured on conventional cell culture plates in media supplemented with calcium or inorganic phosphate to study the effect of these ions independent of the 3D environment created by the particles. Calcium was shown to increase proliferation of cells, whereas both calcium and phosphate positively affected alkaline phosphatase enzyme production. QPCR analysis revealed positive effects of calcium and of inorganic phosphate on the expression of osteogenic markers, in particular bone morphogenetic protein-2 and osteopontin. Higher levels of mineralization were also observed upon exposure to either ion. Effects were similar for cells cultured on composite materials and those cultured in supplemented media, although ion concentrations in the composite cultures were lower. The approach presented here may be a valuable tool for studying the individual effects of a variety of soluble compounds, including bioinorganics, without interference from other material properties.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
L-Ascorbic acid, 99%
Sigma-Aldrich
Dexamethasone, powder, BioReagent, suitable for cell culture, ≥97%
Sigma-Aldrich
L-Ascorbic acid, powder, suitable for cell culture, γ-irradiated
Supelco
Ascorbic Acid, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
L-Ascorbic acid, BioXtra, ≥99.0%, crystalline
Sigma-Aldrich
Dexamethasone, ≥98% (HPLC), powder
Sigma-Aldrich
L-Glutamine, meets USP testing specifications, suitable for cell culture, 99.0-101.0%, from non-animal source
Sigma-Aldrich
L-Ascorbic acid, suitable for cell culture, suitable for plant cell culture, ≥98%
Sigma-Aldrich
Calcium carbonate, ACS reagent, ≥99.0%, powder
Sigma-Aldrich
L-Ascorbic acid, ACS reagent, ≥99%
Sigma-Aldrich
L-Glutamine, ReagentPlus®, ≥99% (HPLC)
Sigma-Aldrich
Calcium carbonate, ACS reagent, chelometric standard, 99.95-100.05% dry basis
SAFC
L-Glutamine
Sigma-Aldrich
Calcium carbonate, ReagentPlus®
Sigma-Aldrich
L-Ascorbic acid, reagent grade, crystalline
Sigma-Aldrich
L-Ascorbic acid, puriss. p.a., ACS reagent, reag. ISO, Ph. Eur., 99.7-100.5% (oxidimetric)
Sigma-Aldrich
Calcium carbonate, powder, ≤50 μm particle size, 98%
Supelco
L-Ascorbic acid, analytical standard
Supelco
Dexamethasone, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Dexamethasone, powder, γ-irradiated, BioXtra, suitable for cell culture, ≥80% (HPLC)
USP
Ascorbic acid, United States Pharmacopeia (USP) Reference Standard
Supelco
L-Ascorbic acid, certified reference material, TraceCERT®
Sigma-Aldrich
L-Ascorbic acid, reagent grade
Ascorbic acid, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
L-Glutamine, BioUltra, ≥99.5% (NT)
Sigma-Aldrich
L-Ascorbic acid, FCC, FG
Sigma-Aldrich
L-Ascorbic acid, meets USP testing specifications
Sigma-Aldrich
Calcium carbonate, BioXtra, ≥99.0%
Sigma-Aldrich
L-Glutamine, γ-irradiated, BioXtra, suitable for cell culture
Sigma-Aldrich
L-Ascorbic acid, puriss. p.a., ≥99.0% (RT)