Document Detail

Influence of hydroxyapatite microstructure on human bone cell response.
MedLine Citation:
PMID:  16628709     Owner:  NLM     Status:  MEDLINE    
Microstructure of calcium phosphate ceramics has been shown to influence long-term in vitro cellular events like proliferation and differentiation, and to favor bone integration in vivo. As long-term cellular events are known to be dependent of early cell adhesion events, we decided to study the in vitro influence of the microstructure of a microporous hydroxyapatite (mHA) and a nonmicroporous hydroxyapatite (pHA) ceramic on serum protein adsorption and SaOs-2 human bone cells attachment after 30 min, 1, 4, and 24 h and cell growth after 96 h. Plastic coverslips were used as controls. Hydroxyapatite composition of mHA and pHA was confirmed by X-ray diffraction and Fourier transform infra-red spectroscopy. The surface energies of ceramics were calculated from contact-angle measurements in di-iodomethane, water or complete culture medium. The total surface energy was 44.8 mJ/m(2) for pHA and 48.7 mJ/m(2) for plastic. The contact-angle measurement was impossible on mHA likely because they displayed 12% of open microporosity, pHA ceramic exhibiting only closed pores (2.5%). Moreover, the roughness amplitude was largely higher on mHA (Sa = 4.35 microm) than on pHA (Sa = 0.065 microm) and plastic (Sa = 0.042 microm). Three different techniques were used to evaluate protein adsorption on the ceramics. SDS-PAGE of desorbed proteins demonstrated that more proteins desorbed from mHA (66.02 microg/m(2)) than from pHA (17.2 microg/m(2)) or plastic (0.08 microg/m(2)). A new method was used to evaluate in situ the quantity of adsorbed total proteins: the temperature-programmed desorption (TPD) analysis coupled with mass spectrometry. The TPD analysis confirmed that 10-fold more proteins adsorbed on mHA compared with those on pHA. A direct immunolabeling on ceramics revealed than more fibronectin and serum albumin adsorbed on microporous ceramic than on dense ceramic. The morphology of SaOs-2 cells was the same on all the substrates after 30 min. At later time points, cell morphology on mHA was radically different than on other surfaces, with the particularity of the cytoplasmic edge that appeared undistinguishable from the surface. Only the extremity of the cells and lamellipodia were visible. Cells seemed like "adsorbed" by the mHA surface, whereas on plastic and pHA surfaces the cells displayed classical aspects of polygonal spreading. The cells displayed on mHA the highest initial attachment potential after 30 min, 1, 4, 24 h but the lower proliferation potential after four days. This study confirms that a microporous ceramic surface can modulate the adsorption of proteins and further the adhesion and proliferation of human bone cells.
Myriam Rouahi; Olivier Gallet; Eric Champion; Joseph Dentzer; Pierre Hardouin; Karine Anselme
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of biomedical materials research. Part A     Volume:  78     ISSN:  1549-3296     ISO Abbreviation:  -     Publication Date:  2006 Aug 
Date Detail:
Created Date:  2006-06-29     Completed Date:  2006-09-12     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101234237     Medline TA:  J Biomed Mater Res A     Country:  United States    
Other Details:
Languages:  eng     Pagination:  222-35     Citation Subset:  IM    
Laboratoire de Recherche sur les Biomatériaux et Biotechnologies, LR2B, quai Robert Masset, Bassin Napoléon, Boulogne sur mer, France.
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MeSH Terms
Biocompatible Materials
Bone and Bones / cytology*,  ultrastructure
Fibronectins / analysis
Microscopy, Electron, Scanning
X-Ray Diffraction
Reg. No./Substance:
0/Biocompatible Materials; 0/Fibronectins; 1306-06-5/Durapatite

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