Effect of silicon doping on bone formation within alumina porous domains

The effect of doping a porous bioinert bioceramic with silicon (Si) on tissue ingrowth, differentiation, and osteogenesis was studied using a rat intramedullary model. Alumina tubes (1.3-mm outer diameter, 0.6-mm inner diameter, 15-mm length) doped with Si at nominal concentrations of 0.5 and 5.0 mol % were implanted into femoral medullary canals of female rats for 16 weeks. Tissue formation within the tubes was determined by histology and histomorphometry. Addition of 0.5 mol % Si to alumina stimulated cellular activity at the bone-ceramic interface and impaired osteogenic maturation within the tubes. In contrast, osteogenesis was enhanced in the 5.0 mol % Si-doped alumina tubes. It is considered that effect of Si is related to surface chemistry rather than microstructure. This work demonstrates that doping a bioinert ceramic with small amounts of Si can significantly alter tissue ingrowth, differentiation, and osteogenesis within a porous implant.     

Bone formation process in porous calcium carbonate and hydroxyapatite.

This study determined the bone formation in porous calcium carbonate (CC) and porous hydroxyapatite (HA) in ectopic sites. The bone formation stimulus was derived from bone marrow cells. CC and HA in the shape of disks were implanted with or without rat marrow cells into subcutaneous sites of syngeneic rats. The CC and HA had identical microstructure: pore size was 190-230 microns, porosity was 50-60% and they were fully interconnected. Bone did not form in any implants without marrow cells (disks themselves), whereas bone consistently formed in the pores of all implants with marrow cells after 4 weeks. The bone formation of both CC and HA occurred initially on surface of the pore regions and progressed toward the center of the pore. Scanning electron microscopy and electron-probe microanalysis revealed a continuum of calcium at the interfaces of both bone/CC and bone/HA implants. These results indicate that the bone formation in calcium carbonate derived from marine corals is comparable to the bioactive hydroxyapatite.     

Bone formation analysis: effect of quantification procedures on the study outcome

Quantification of the amount of newly formed bone is an essential part of bone regeneration studies. Histomorphometry, based on histological sections of plastic-embedded specimens, is the most frequently applied technique in this assessment. Before performing image analysis, a specific region of interest (ROI) has to be determined. Based on the histological procedure, different areas within the ROI can be discriminated and assigned to relevant tissue structures. However, in literature not much attention is paid to the effect of the histological procedures on the final outcome of the histomorphometrical measurements on bone regeneration. In this study, the histomorphometrical bone formation of the intramedullary cavity of the guinea pig tibia, filled with calcium phosphate cement, was quantified in plastic-embedded and paraffin-embedded specimens and in specimens analyzed with scanning electron microscopy in the backscattering mode (SEM-BS). The data showed that the histological procedure significantly affected the measured bone amount. Therefore, it is recommended that scaffold characteristics are carefully considered in selecting a proper technique for the analysis of bone formation in bone tissue engineering studies. The results of this study identified high-resolution SEM-BS and elastic van Gieson staining of decalcified histological sections as recommendable techniques for evaluating bone formation.     

Alginate-controlled formation of nanoscale calcium carbonate and hydroxyapatite mineral phase within hydrogel networks

A one-step method was used to make nanostructured composites from alginate and calcium carbonate or calcium phosphate. Nanometer-scale mineral phase was successfully formed within the gel network of alginate gel beads, and the composites were characterized. It was found that calcite was the dominating polymorph in the calcium carbonate mineralized beads, while stoichiometric hydroxyapatite was formed in the calcium phosphate mineralized beads. A combination of electron microscopy, Fourier-transform infrared spectroscopy, thermogravimetric analysis and powder X-ray diffraction showed that alginate played an active role in controlling mineral size, morphology and polymorphy. For the calcium phosphate mineralized beads, alginate was shown to modulate stoichiometric hydroxyapatite with low crystallinity at room temperature, which may have important applications in tissue engineering. The results presented in this work demonstrate important aspects of alginate-controlled crystallization, which contributes to the understanding of composite material design.     

Health status of persons occupationally exposed to chromium, nickel, manganese and polycyclic aromatic hydrocarbons

Occupational environment monitoring and biological-medical monitoring of persons professionally exposed to welding fumes have been performed. Chromium, manganese and polycyclic aromatic hydrocarbons in welding fumes represents an important health risk. Pollutant concentrations found in metal welding fumes represented only fractions of those acceptable ones. Polycyclic aromatic hydrocarbons have been reached the concentration found in a busy road crossing in Hradec Králové (compared with these as in Czech Republic no maximum acceptable levels for PAHs having been declared). Family, personal and occupational history have been taken. Health state including total haematological count, biochemical and cytogenetical changes of 19 stainless steel welders were checked-up. The level of mercapturates in urine were examined as well. The data were statistically compared with those of non exposed (control group). No changes witnessing the above mentioned risk factors influence on the haematological, biochemical and cytogenetical findings were ever proved. In conclusion, our results did not confirm an increased professional risk in this group of welders.     

Bone formation in calvarial defects of Sprague-Dawley rats by transplantation of calcium phosphate glass

The purpose of this study was to investigate the bone-regenerative effect of calcium phosphate glass in vivo. We prepared amorphous calcium phosphate glass powder having a mean particle size of 400 microm in the system CaO-CaF2-P2O5-MgO-ZnO. Calvarial critical-sized defects (8 mm) were created in 60 male Sprague-Dawley rats. The animals were divided into an experimental group and control group of 30 animals each. Each defect was filled with a constant weight of 0.5 g calcium phosphate glass powder mixed with saline. As a control, the defect was left empty. The rats were sacrificed 2, 4, or 8 weeks postsurgery, and the results evaluated using radiodensitometric and histological studies; they were also examined histomorphometrically. When the calcium phosphate glass powders with 400-microm particles were grafted, the defects were nearly completely filled with new-formed bone in a clean healing condition after 8 weeks. It was observed that the prepared calcium phosphate glass enhanced new bone formation in the calvarial defect of Sprague-Dawley rats and could be expected to have potential for use as a hard tissue regeneration material.     

Bone tissue formation with human mesenchymal stem cells and biphasic calcium phosphate ceramics: The local implication of osteoclasts and macrophages

Human mesenchymal stem cells (hMSC) have immunomodulative properties and, associated with calcium phosphate (CaP) ceramics, induce bone tissue repair. However, the mechanisms of osteoinduction by hMSC with CaP are not clearly established, in particular the role of osteoclasts and macrophages. Biphasic calcium phosphate (BCP) particles were implanted with or without hMSC in the paratibial muscles of nude mice. hMSC increased osteoblastic gene expression at 1 week, the presence of macrophages at 2 and 4 weeks, osteoclastogenesis at 4 and 8 weeks, and osteogenesis at 4 and 8 weeks. hMSC disappeared from the implantation site after 2 weeks, indicating that hMSC were inducers rather than effectors of bone formation. Induced blockage of osteoclastogenesis by anti-Rankl treatment significantly impaired bone formation, revealing the pivotal role of osteoclasts in bone formation. In summary, hMSC positively influence the body foreign reaction by attracting circulating haematopoietic stem cells and inducing their differentiation into macrophages M1 and osteoclasts, thus favouring bone formation.     

The effect of different powder particle size on mechanical properties of sintered alumina, resin- and glass-infused alumina

In this study, the compaction and sintering behavior of fine alumina powders of different particle sizes and the effect of matrix particle size on biaxial strength and fracture toughness of infused matrices were investigated. Three different alumina powders, In-Ceram alumina, A16SG, and RC172 were selected, representing a range of particle size and shape. RC172 and A16SG were dry-pressed. In-Ceram alumina was slip-cast following manufacturer's recommendations. Dry-pressed ceramic blocks were sectioned into disks with a thickness of 1.5-mm. Uninfused disks were sintered at four temperatures between 1250 degrees C and 1400 degrees C. For glass or resin infused specimens, alumina disks were sintered at 1250 degrees C for 2 h and separated into two groups for glass infusion and resin (UDMA/TEGDMA) infusion. Disks were tested for biaxial flexural strength with a universal testing machine (Instron) at 0.5-mm/min crosshead speeds. One-way ANOVA and Duncan's multiple range tests revealed that alumina disks with different smaller particle sizes have significantly higher biaxial strength (p < 0.05). The strength of the alumina matrix was greatly increased by glass and resin infusion. The biaxial strength of resin-infused alumina increased as particle size decreased, whereas strength of glass-infused alumina was constant.     

The fabrication and biochemical evaluation of alumina reinforced calcium phosphate porous implants

Alumina reinforced calcium phosphate porous implants were manufactured to improve the mechanical strength while maintaining the bioactivity of calcium phosphate ceramics. The alumina porous bodies, which provided the mechanical strength, were fabricated by a polyurethane sponge method and multiple coating techniques resulted in the porous bodies with a 90-75% porosity and a compressive strength of up to approximately 6MPa. The coating of hydroxyapatite (HAp) or tricalcium phosphate (beta-TCP) was performed by dipping the alumina porous bodies into calcium phosphate ceramic slurries and sintering the specimens. The fairly strong bonding between the HAp or TCP coating layer and the alumina substrate was obtained by repeating the coating and sintering processes. The biochemical evaluations of the porous implants were conducted by in vitro and in vivo tests. For in vitro test, the implants were immersed in Ringer's solution and the release of Ca and P ions were detected and compared with those of calcium phosphate powders. For in vivo test, the porous bodies were implanted into mixed breed dogs and bone mineral density measurements and histological studies were conducted. The alumina reinforced HAp porous implants had a higher strength than the HAp porous implants and exhibited a similar bioactivity and osteoconduction property to the HAp porous implants.     

Women with fibromyalgia have lower levels of calcium, magnesium, iron and manganese in hair mineral analysis

Little is known about hair mineral status in fibromyalgia patients. This study evaluated the characteristics of hair minerals in female patients with fibromyalgia compared with a healthy reference group. Forty-four female patients diagnosed with fibromyalgia according to the American College of Rheumatology criteria were enrolled as the case group. Age and body mass index-matched data were obtained from 122 control subjects enrolled during visit for a regular health check-up. Hair minerals were analyzed and compared between the two groups. The mean age was 43.7 yr. General characteristics were not different between the two groups. Fibromyalgia patients showed a significantly lower level of calcium (775 μg/g vs 1,093 μg/g), magnesium (52 μg/g vs 72 μg/g), iron (5.9 μg/g vs 7.1 μg/g), copper (28.3 μg/g vs 40.2 μg/g) and manganese (140 ng/g vs 190 ng/g). Calcium, magnesium, iron, and manganese were loaded in the same factor using factor analysis; the mean of this factor was significantly lower in fibromyalgia group in multivariate analysis with adjustment for potential confounders. In conclusion, the concentrations of calcium, magnesium, iron, and manganese in the hair of female patients with fibromyalgia are lower than of controls, even after adjustment of potential confounders.     

Stimulatory effect of zinc-releasing calcium phosphate implant on bone formation in rabbit femora

Although hydroxyapatite (HAP) and tricalcium phosphate (TCP) are currently used as bone graft substitutes or coatings on metallic prostheses because of their excellent biocompatibility and osteoconductivity, they do not stimulate bone formation or inhibit bone resorption. Zinc, an essential trace element in many animals, has a direct specific proliferative effect on osteoblastic cells and has a potent and selective inhibitory effect on osteoclastic bone resorption in vitro. Therefore, zinc-containing beta-tricalcium phosphate (ZnTCP) ceramics and composite ceramics of ZnTCP and HAP (ZnTCP/HAP) were implanted in the femora of New Zealand White rabbits for 4 weeks to promote bone formation. The implants were sintered ceramics with zinc contents of 0 (control), 0.063, 0.316 and 0.633 wt %. Histological and histomorphometrical investigation of the undecalcified sections revealed an increase by 51% (p =.0509) in the area of newly formed bone around the ZnTCP/HAP implants of 0. 316 Zn wt % compared with the control. Plasma zinc concentration was unchanged. An increased bone resorption on the endosteal surface was observed when ZnTCP and ZnTCP/HAP of 0.633 Zn wt % were implanted. To promote bone formation, the optimum zinc content of the calcium phosphate ceramics was therefore 0.316 wt %.     

Bone formation by rat bone marrow cells cultured on titanium fiber mesh: effect of in vitro culture time

The objective of this study was to examine the effect of cell culture time on bone formation by rat bone marrow cells seeded in titanium fiber mesh. As a seeding technique, a high cell suspension was used (3 x 10(6) cells/mL). Therefore, 30 meshes were repeatedly rotated in a 10 mL tube (containing 30 x 10(6) cells) on a rotation plate (2 rpm) for 3 h. Osteogenic cells were cultured for 1, 4, and 8 days on titanium fiber mesh and finally implanted subcutaneously in rats. Meshes without cells were also implanted subcutaneously in rats. DNA and scanning electron microscopy (SEM) analyses and calcium measurements determined cellular proliferation and differentiation during the in vitro incubation period of the mesh implants. Four weeks after implant insertion, the animals were sacrificed. The implants, with their surrounding tissue, were retrieved and prepared for histologic evaluation and calcium measurements. DNA analysis of the in vitro experiment showed a lag phase from day 1 through day 4, but a 42% increase in DNA between days 4 and 8. SEM and calcium measurements indicated an increase in calcium from day 1 to day 4, yet only a small but significant increase from days 4 to 8. Histologic analysis demonstrated that bone was formed in all day 1 and day 4 implants, and that the bone-like tissue was present uniformly through the meshes. The bony tissue was morphologically characterized by osteocytes embedded in a mineralized matrix, with a layer of osteoid and osteoblasts at the surface. The day 8 implants showed only calcium phosphate deposition in the titanium fiber mesh. Calcium measurements of the implants revealed that calcification in day 1 implants was significantly higher (p < 0.05) compared to day 4 and day 8 implants. No significant difference in calcium content existed between day 4 and day 8 implants. On the basis of our results, we conclude that 1) bone formation was generated more effectively in osteogenic cells by a short culture time after seeding in titanium fiber mesh; 2) dynamic cell seeding is probably more effective than static cell seeding; and 3) selection of the right cells from the heterogenous bone marrow population remains a problem.     

Bone tissue growth enhancement by calcium phosphate coatings on porous titanium alloys: the effect of shielding metal dissolution product.

The possible mechanism of minimization of prosthesis-derived bone growth inhibitors by shielding of the metal and the reduction, if not elimination, of the associated metal dissolution was investigated. Titanium, aluminium and vanadium release rates were determined in vitro for Ti alloy specimens both with and without a calcium phosphate coating. Ti orderly oriented wire mesh (OOWM) porous coatings on Ti-6Al-4V substrates were used as the metal specimens. Half of the specimens were coated with a 75 microns calcium phosphate ceramic (CPC coating). Seven reference (OOWM) and seven coated (OOWM-CPC) specimens were immersed and placed along with seven control solutions for various periods in an incubator maintained at 37 degrees C and 5% CO2 - air atmosphere. Whereas the reference solutions showed a Ti release increasing as a function of time, the solutions that had the CPC-coated specimens contained no measurable amounts of titanium. The Al in solution around the CPC-coated specimens was significantly greater than the concentration around non-coated specimens. The Al, however, did not increase significantly with time, at least up to 4 wk immersion. The ceramic coating had a small beneficial effect on V concentration. In the absence of a significant adverse effect of Ti on local bone tissue formation, we focus on the Al data of our study. The possible adverse effect of this element is well documented. The calcium phosphate coating produced a significant increase of biological fixation, yet at the same time a greater Al release into solution, calling into question the significance of CPC coating in shielding adverse metal passive dissolution to explain enhanced bone growth [corrected].     

Bone formation in cardiac valves: a histopathological study of 128 cases

In a series of 1,177 patients with surgical valve resection, there were 128 (10.9%) patients with the histological finding of bone and/or cartilage formation (bony metaplasia; heterotopic ossification) in the excised valve (119 aortic, 9 mitral). The incidence was higher in men (12.8%) than in women (8.5%). The bony metaplasia was always associated with valvular calcification. The metaplastic changes were encountered in all types of calcific aortic valve disease (senile, bicuspid valve, post-rheumatic). The marrow of the heterotopic lamellar bone was mostly composed of adipose tissue. Frequently, it showed chronic inflammation with polyclonal plasmocytic infiltration. Complete haematopoiesis was present only rarely.     

Bone mass regulation by leptin: a hypothalamic control of bone formation

Bone mass is maintained constant between puberty and menopause by the balance between osteoblasts and osteoclasts activity. The existence of a hormonal control of osteoblast activity has been speculated for years by analogy to osteoclast biology. Through the search for such humoral signal(s) regulating bone formation, leptin has been identified as a powerful inhibitor of bone formation. Furthermore, by means of intracerebroventricular infusion of leptin, it has been shown that the effect of this adipocyte-derived hormone on bone is mediated via a brain relay, like all its other functions. Subsequent studies have led to the identification of hypothalamic neurons involved in leptin's antiosteogenic function. In addition, it has been shown that those neurons or neuronal pathways are distinct from neurons responsible for the regulation of energy metabolism. Finally, the peripheral mediator of leptin's antiosteogenic function has been identified as being the sympathetic nervous system. Catecholamine-deficient mice have a high bone mass and sympathomimetics administered to mice decreased bone formation and bone mass. Conversely, beta-blockers increased bone formation and bone mass and blunt the bone loss induced by ovariectomy.