RESUMO
Microcracks are present in bone and can result from fatigue damage due to repeated, cyclically applied stresses. From a mechanical point, microcracks can dissipate strain energy at the advancing tip of a crack to improve overall bone toughness. Physiologically, microcracks are thought to trigger bone remodeling. Here, we examine the effect of microcracks specifically on osteoblasts, which are bone-forming cells, by comparing cell responses on microcracked versus non-microcracked hydroxyapatite (HA) specimens. Osteoblast attachment was found to be greater on microcracked HA specimens (p<0.05). More importantly, we identified the preferential alignment of osteoblasts in the direction of the microcracks on HA. Cells also displayed a preferential attachment that was 75 to 90 µm away from the microcrack indent. After 21 days of culture, osteoblast maturation was notably enhanced on the HA with microcracks, as indicated by increased alkaline phosphatase activity and gene expression. Furthermore, examination of bone deposition by confocal laser scanning microscopy indicated preferential mineralization at microcrack indentation sites. Dissolution studies indicate that the microcracks increase calcium release, which could contribute to osteoblast responses. Our findings suggest that microcracks signal osteoblast attachment and bone formation/healing.
Assuntos
Osteoblastos/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Células 3T3-L1 , Animais , Remodelação Óssea/efeitos dos fármacos , Osso e Ossos/efeitos dos fármacos , Osso e Ossos/fisiologia , Diferenciação Celular/efeitos dos fármacos , Durapatita/farmacologia , Camundongos , Osteoblastos/citologia , Tamanho da Partícula , Manejo de Espécimes , Propriedades de Superfície , Alicerces Teciduais/química , Difração de Raios XRESUMO
This study analyzes data from 206 CaP specimens (68 HA, 70 BCP, and 68 beta-TCP) fractured via biaxial flexure testing. Specimens were divided into four groups: (a) Group I, dry; (b) Group II, wet (day 0, immersion time approximately 5-10 s); (c) Group III, after immersion in media for 21 days (day 21); and (d) Group IV, after culturing osteoblasts (OBs) on the surface for 21 days (day 21 with cells). X-ray diffraction verified the presence of minor second phases in HA and beta-TCP while BCP was a biphasic mixture of HA and beta-TCP with minor phases present. The statistical significance (p < 0.05) of differences in the measured biaxial flexure fracture strength, S, between groups was assessed via one-way ANOVA with Tukey's test. Also, a two-parameter Weibull analysis assessed the mechanical reliability of each group. Osteoblasts increase the biaxial flexure fracture strength in a statistically significant way compared to both the HA discs in Groups II and III. Scanning electron microscope examination revealed grain boundary grooving on the sintered surfaces and with thermal expansion anisotropy, likely leads to the observed rapid strength decline upon exposure to media found in Groups II, III and IV.
Assuntos
Fosfatos de Cálcio , Teste de Materiais , Osteoblastos/citologia , Engenharia Tecidual , Animais , Linhagem Celular , Cerâmica , Durapatita , Mecânica , Camundongos , Microscopia EletrônicaRESUMO
Hydroxyapatite (HAp) is the major mineral constituent of bone, and as such, the dielectric properties of HAp are of interest because electromagnetic fields have been shown to accelerate healing in bone fractures. In addition, an interest in the dielectric properties of HAp stems from the suggestion that electrically insulating HAp coatings might be used on implantable devices. In this study, the dielectric constant of polycrystalline hexagonal HAp was measured at nine different frequencies, from 45 kHz to 7.3 MHz for relative porosities ranging from 0.05 to 0.42. At a fixed frequency, the decrease in k as a function of increasing porosity is described well by an exponential function of porosity such that k = k(0)exp(-bP), where k(0) is the dielectric constant at zero porosity and b is a constant. In addition, the entire data set of 108 data points (representing the 12 specimens of differing porosity measured at each of the nine frequencies) was fit to a candidate function formed from the product k(0)exp(-bP) and a simple expression relating frequency to the dielectric constant. The candidate function fit the data relatively well, with a coefficient of determination of 0.91.