title: The effects of Ca2SiO4-Ca3(PO4)2 ceramics on adult human mesenchymal stem cell viability, adhesion, proliferation, differentiation and function.
abstract: Bioceramic samples with osteogenic properties, suitable for use in the regeneration of hard tissue, were synthesized. The materials consisting of α-tricalcium phosphate (αTCP) and also αTCP doped with either 1.5 wt.% or 3.0 wt.% of dicalcium silicate (C2S) in the system Dicalcium Silicate-Tricalcium Phosphate (C2S-TCP) were obtained by solid state reaction. All materials were composed of a single phase, αTCP in the case of a pure material, or solid solution of C2S in αTCP (αTCPss) for the doped αTCP. Viability, proliferation and in vitro osteoinductive capacity were investigated by seeding, adult mesenchymal stem cells of human origin (ahMSCs) which were CD73(+), CD90(+), CD105(+), CD34(-) and CD45(-) onto the 3 substrates for 30 days. Results show a non-cytotoxic effect after applying an indirect apoptosis test (Annexin V/7-AAD staining), so ahMSCs adhered, spread, proliferated and produced extracellular matrix (Heparan-sulfate proteoglycan (HS) and osteopontin (OP)) on all the ceramics studied. Finally, the cells lost the cluster differentiation marker expression CD73, CD90 y CD105 characteristic of ahMSCs and they showed an osteoblastic phenotype (Alkaline phosphatase activity (ALP), Osteocalcin production (OC), Collagen type I expression (Col-I), and production of mineralization nodules on the extracellular matrix). These observations were more evident in the αTCP ceramic doped with 1.5 wt.% C2S, indicating osteoblastic differentiation as a result of the increased concentration of solid solution of C2S in αTCP (αTCPss). Overall, these results suggest that the ceramics studied are cytocompatible and they are able to induce osteoblastic differentiation of undifferentiated ahMSCs.
authors.affiliation: Instituto de Bioingeniería, Universidad Miguel Hernández, Avda. Universidad s/n, 03202 Elche, Alicante, Spain. piedad@umh.es
authors.forename: Piedad N
authors.initials: PN
authors.lastname: De Aza
authors.forename: David
authors.initials: D
authors.lastname: García-Bernal
authors.forename: Francesca
authors.initials: F
authors.lastname: Cragnolini
authors.forename: Pablo
authors.initials: P
authors.lastname: Velasquez
authors.forename: Luis
authors.initials: L
authors.lastname: Meseguer-Olmo
mesh_terms: D000328:Adult; D053687:Adult Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology / Q000648:ultrastructure; D000469:Alkaline Phosphatase / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects; D002118:Calcium / Q000032:analysis; D017610:Calcium Compounds / Q000494:pharmacology*; D002130:Calcium Phosphates / Q000494:pharmacology*; D002448:Cell Adhesion / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002516:Ceramics / Q000494:pharmacology*; D055598:Chemical Phenomena / Q000187:drug effects; D003470:Culture Media / Q000737:chemistry; D006801:Humans; D008422:Materials Testing; D055595:Mechanical Phenomena / Q000187:drug effects; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology / Q000648:ultrastructure; D015675:Osteocalcin / Q000378:metabolism; D010758:Phosphorus / Q000032:analysis; D017640:Silicates / Q000494:pharmacology*; D012825:Silicon / Q000032:analysis
chemical_list: D017610:Calcium Compounds; D002130:Calcium Phosphates; D003470:Culture Media; D017640:Silicates; D015675:Osteocalcin; D010758:Phosphorus; D000469:Alkaline Phosphatase; C018392:tricalcium phosphate; C031293:calcium silicate; D002118:Calcium; D012825:Silicon
keywords: Adult human mesenchymal stem cells; Biomedical applications; Electron microscopy; Powder-solid state reaction; Silicate; Tricalcium phosphate