In vitro enhancement of SAOS-2 cell calcified matrix deposition onto radio frequency magnetron sputtered bioglass-coated titanium scaffolds.

In bone tissue engineering, bioglass coating of titanium (Ti) scaffolds has drawn attention as a method to improve osteointegration and implant fixation. In this in vitro study, bioactive glass layers with an approximate thickness of 1 microm were deposited at 200 degrees C onto a three-dimensional Ti-6Al-4V scaffold using a radio frequency (r.f.) magnetron sputtering system. After incubation with SAOS-2 human osteoblasts, in comparison with the uncoated scaffolds, the bioglass-coated scaffolds showed a twofold increase in cell proliferation (p < 0.05) up to 68.4 x 10(6), and enhanced the deposition of extracellular matrix components such as decorin, fibronectin, osteocalcin, osteonectin, osteopontin, and type-I and -III collagens (p < 0.05). Calcium deposition was twofold greater on the bioglass-coated scaffolds (p < 0.05). The immunofluorescence related to the preceding bone matrix proteins and calcium showed their colocalization to the cell-rich areas. Alkaline phosphatase activity increased twofold (p < 0.001) and its protein content was threefold higher with respect to the uncoated sample. Quantitative reverse transcriptase-polymerase chain reaction analysis revealed upregulated transcription specific for type-I collagen and osteopontin (p < 0.001). All together, these results demonstrate that the bioglass coating of the three-dimensional Ti scaffolds by the r.f. magnetron sputtering technique determines an in vitro increase of the bone matrix elaboration and may potentially have a clinical benefit.

The response of bone to nanocrystalline hydroxyapatite-coated Ti13Nb11Zr alloy in an animal model.

An in vivo study was carried out on uncoated and hydroxyapatite (HA)-coated nanostructured Ti13Nb11Zr alloy in comparison with high-grade Ti6Al4V, to investigate the effect of the different surfaces on osteointegration rate. A highly effective method to obtain a fast biomimetic deposition of a thin layer of nanocrystalline HA was applied to coat both substrates. Cylindrical pins were implanted in rabbit cortical bone and evaluated at 4 and 12 weeks by histomorphometry and microhardness tests. The results confirmed the ability of the slightly supersaturated Ca/P solution to induce a fast deposition of nanocrystalline HA on Ti alloys' surfaces. HA-coated Ti13Nb11Zr had the highest osteointegration rate at 4 and 12 weeks. Both HA-coated surfaces showed an affinity index significantly higher than those of native surfaces at 4 weeks (Ti13Nb11Zr+HA: 37%; Ti6Al4V+HA: 26%). Microhardness test showed a significantly higher bone mineralization index of HA-coated Ti13Nb11Zr in comparison with that of HA-coated Ti6Al4V surface. The study suggests that the HA coating on both alloys enhances bone response around implants and that there is a synergic effect of Ti-Nb-Zr alloy with the HA coating on bone remodeling and maturation.

Amidic alginate hydrogel for nucleus pulposus replacement.

Degeneration of intervertebral discs is the most common cause of back pain. The first phase of this degenerative process involves the nucleus pulposus (NP). A rapid recovery of this structure can prevent further degradation of the annulus fibrosus. A new amidic derivative of alginate (AAA) was developed to obtain a polysaccharide possessing some of the physical-chemical properties of Hyal (i.e. viscosity) without losing the rigidity of the native alginate structure. The modified polysaccharide was crosslinked using 1.3 diaminopropane as crosslinking agent. The hydrogel obtained was characterized in terms of water uptake and rheological behavior. In particular, the viscoelastic behavior of the hydrogel was determined in shear stress under dynamic conditions and compared with the behavior of nondegenerated human lumbar NP. We then assessed the effect of the AAA hydrogel on NHC (Normal Human Chondrocyte) cell viability and on the production of important extracellular matrix factors, such as glycosaminoglycans and Type II collagen. In conclusion, the results achieved in this study demonstrated that the amidic alginate-based scaffold is a promising material to be utilized in the replacement of NP.

In vitro differentiation of human mesenchymal stem cells on Ti6Al4V surfaces.

Long-term stability of arthroplasty prosthesis depends on the integration between the bone tissue and the implanted biomaterials, which requires the contribution of osteoblastic precursors and their continuous differentiation into the osteoblastic phenotype. Classically, these interactions are tested in vitro using mesenchymal stem cells (MSCs) isolated and ex vivo expanded from bone marrow aspirates. Human adipose tissue-derived stromal cells (AMSCs) may be a more convenient source of MSCs, according to their abundance and accessibility, but no data are available on their in vitro interactions with hard biomaterials. The aim of this work is to compare the osteogenic potential of human AMSCs and bone marrow-derived MSCs (BMMSCs) and to evaluate their response to Ti6Al4V alloy in terms of adhesion, proliferation and differentiation features, using the human osteosarcoma cell line SaOS-2 for comparison. The overall results showed that AMSCs have the same ability to produce bone matrix as BMMSCs and that Ti6Al4V surfaces exhibit an osteoinductive action on AMSCs, promoting their differentiation into functional osteoblasts and increasing bone formation. In conclusion, adipose tissue is a promising autologous source of osteoblastic cells with important clinical implications for bone tissue engineering.

Nanocrystalline hydroxyapatite coatings on titanium: a new fast biomimetic method.

We obtained a fast biomimetic deposition of hydroxyapatite (HA) coatings on Ti6Al4V substrates using a slightly supersaturated Ca/P solution, with an ionic composition simpler than that of simulated body fluid (SBF). At variance with other fast deposition methods, which produce amorphous calcium phosphate coatings, the new proposed composition allows one to obtain nanocrystalline HA. Soaking in supersaturated Ca/P solution results in the deposition of a uniform coating in a few hours, whereas SBF, or even 1.5SBF, requires 14 days to deposit a homogeneous coating on the same substrates. The coating consists of HA globular aggregates, which exhibit a finer lamellar structure than those deposited from SBF. The extent of deposition increases on increasing the immersion time. Transmission electron microscope (TEM) images recorded on the material detached from the coating show that the deposition is constituted of thin nanocrystals. Electron diffraction (ED) patterns recorded from most of the crystals exhibit the presence of rings, which can be indexed as reflections characteristic of HA. Furthermore, several HA single-crystal spot ED images were obtained from individual crystals.