title: Comparative study of mesenchymal stem cells osteogenic differentiation on low-temperature biomineralized nanocrystalline carbonated hydroxyapatite and sintered hydroxyapatite.
abstract: Hydroxyapatite with different characteristics in terms of morphology and chemistry were prepared via conventional sintering and low temperature biomimetic mineralization methods. The biomineralization route introduced nanocrystalline carbonate-substituted hydroxyapatite (n-CHA) with needle-like crystals ranging 20-30 nm whereas sintered HA (S-HA) comprised of polygonal grains ranging 2-5 μm. The response of fibroblastic cells was investigated using the extract of the samples whereas Wistar rat-derived mesenchymal stem cells (MSCs) were evaluated on top of each sample while maintaining in an osteogenic-free medium. The proliferation, activity, and morphology of adherent MSCs were determined at different culturing periods. The osteogenic differentiation of MSCs was also assayed by determining expression of runx2, osteonectin, osteopontin, and osteocalcin genes using real time-PCR analysis. The fibroblastic cells exhibited better proliferation rate at the presence of n-CHA compared to S-HA. Furthermore, the MSCs attached and spread well on both n-CHA and S-HA with better proliferation rate and alkaline phosphatase activity on n-CHA. Interestingly, the osteogenic differentiation of MSCs on n-CHA was confirmed by the expression of bone specific proteins whereas poor expression of these proteins was detected for the cells on S-HA. The results showed that the role of morphology, crystallinity, and chemistry of hydroxyapatite is crucial for osteogenesis differentiation of MSCs. The results predict osteoinductivity of n-CHA, because MSCs differentiation occurred at the absence of osteogenic medium. However, in vivo data are also required to support this suggestion.
authors.affiliation: Nanotechnology and Advanced Materials Department, Materials and Energy Research Center, Karaj, P.O. Box: 31787/316, Iran.
authors.forename: Saeed
authors.initials: S
authors.lastname: Hesaraki
authors.forename: Hamid
authors.initials: H
authors.lastname: Nazarian
authors.forename: Milad
authors.initials: M
authors.lastname: Pourbaghi-Masouleh
authors.forename: Shokoufeh
authors.initials: S
authors.lastname: Borhan
mesh_terms: D000818:Animals; D040761:Biomimetic Materials / Q000737:chemistry; D018786:Bone Substitutes / Q000737:chemistry*; D002113:Calcification, Physiologic; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics; D017886:Durapatite / Q000737:chemistry*; D015870:Gene Expression; D008422:Materials Testing; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D018625:Microscopy, Atomic Force; D008855:Microscopy, Electron, Scanning; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D015675:Osteocalcin / Q000235:genetics; D010012:Osteogenesis; D015676:Osteonectin / Q000235:genetics; D053495:Osteopontin / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D018786:Bone Substitutes; D050659:Core Binding Factor Alpha 1 Subunit; D015676:Osteonectin; C493717:Runx2 protein, rat; C506897:Spp1 protein, rat; D015675:Osteocalcin; D053495:Osteopontin; D017886:Durapatite
keywords: biocompatibility/hard tissue; biomimetic; calcium phosphate(s); hydroxyapatite; osteogenesis