A new multiphase calcium phosphate graft material improves bone healing-An in vitro and in vivo analysis.

This study aims to evaluate the potential of a novel biomaterial synthesized from amorphous calcium phosphate (ACP), octacalcium phosphate (OCP), and hydroxyapatite (HA) to repair critical-sized defects (CSD) in rabbit calvaria. In vitro analyses of cell viability, cell proliferation, formation of mineral nodules, and cell differentiation using qPCR were performed for comparing experimental calcium phosphate (ECP), deproteinized bovine bone (DBB), and beta-tricalcium phosphate (ß-TCP). Bilateral CSDs were created in 45 rabbit calvaria. Six groups were evaluated: ECP, ECP + fibrin sealant (ECP + S), coagulum, autogenous bone, DBB, and ß-TCP. Euthanasia was performed at 2, 4, and 8 weeks, followed by micro-computed tomography and histological and immunohistochemical analyses. Results from in vitro analyses revealed similar biocompatibility for all tested materials and a tendency for higher gene expression of some bone markers in the ECP group than in ß-TCP and DBB groups at 7 days. In contrast to that in DBB and ß-TCP groups, ECP displayed growing bone volume over total volume percentage (BV/TV%) with time in vivo. Histological analysis revealed a greater number of giant cells and reduced size of grafted particles in ECP during all periods of analysis. RUNX-2 expression was statistically lower in ECP than DBB at 2 and 4 weeks. Despite no statistical significance, ECP presented the highest absolute values for ALP-expression at 2, 4, and 8 weeks compared with other groups. Together, our findings indicate that a combination of the ACP, OCP, and HA phases into ECP is beneficial and promising for bone regeneration.

Sol-gel based calcium phosphates coatings deposited on binary Ti-Mo alloys modified by laser beam irradiation for biomaterial/clinical applications.

Ti-15Mo alloy samples were irradiated by pulsed Yb: YAG laser beam under air and atmospheric pressure. Calcium phosphate coatings were deposited on the irradiated surfaces by the sol-gel method. The sol was prepared from the precursors Ca (NO3)2.4H2 O and H3 PO4. The modified surfaces were submitted to heat treatment conditions at 350 and 600 °C. The results showed that the two conditions established have a sufficient energy to promote ablation on the laser beam irradiated surfaces. Likewise, it has been demonstrated the processes of fusion and fast solidification from the laser beam irradiation, under ambient atmosphere, inducing the formation of stoichiometric TiO2 and non-stoichiometric titanium oxides, including Ti3O5, TiO, Ti3O and Ti6O with different oxide percentages depending on the fluency used. Besides that, laser modification has allowed a clean and reproducible process, providing no traces of contamination, an important feature for clinical applications. The physico-chemical and morphological properties indicated the formation of a mixture of phases: calcium pyrophosphate, hydroxyapatite and ß-TCP for the procedure (PA: calcination temperature), whereas HA (hydroxyapatite) and ß-TCP (tricalcium phosphate) were obtained by the procedure (PB: calcination temperature). Therefore, it was possible to obtain a Ti-15Mo alloy surface consisted on calcium phosphate ceramics of biological interest using the procedure (PB). Thus, the laser beam irradiation associated to bioactive coatings of calcium phosphates of biological interest have shown to be promising and economically feasible for use in dental and orthopedic implants.

Skeletal stem cell and bone implant interactions are enhanced by LASER titanium modification.

PURPOSE: To evaluate the osteo-regenerative potential of Titanium (Ti) modified by Light Amplification by Stimulated Emission of Radiation (LASER) beam (Yb-YAG) upon culture with human Skeletal Stem Cells (hSSCs(1)). METHODS: Human skeletal cell populations were isolated from the bone marrow of haematologically normal patients undergoing primary total hip replacement following appropriate consent. STRO-1(+) hSSC(1) function was examined for 10 days across four groups using Ti discs: i) machined Ti surface group in basal media (Mb(2)), ii) machined Ti surface group in osteogenic media (Mo(3)), iii) LASER-modified Ti group in basal media (Lb(4)) and, iv) LASER-modified Ti group in osteogenic media (Lo(5)). Molecular analysis and qRT-PCR as well as functional analysis including biochemistry (DNA, Alkaline Phosphatase (ALP(6)) specific activity), live/dead immunostaining (Cell Tracker Green (CTG(7))/Ethidium Homodimer-1 (EH-1(8))), and fluorescence staining (for vinculin and phalloidin) were undertaken. Inverted, confocal and Scanning Electron Microscopy (SEM) approaches were used to characterise cell adherence, proliferation, and phenotype. RESULTS: Enhanced cell spreading and morphological rearrangement, including focal adhesions were observed following culture of hSSCs(1) on LASER surfaces in both basal and osteogenic conditions. Biochemical analysis demonstrated enhanced ALP(6) specific activity on the hSSCs(1)-seeded on LASER-modified surface in basal culture media. Molecular analysis demonstrated enhanced ALP(6) and osteopontin expression on titanium LASER treated surfaces in basal conditions. SEM, inverted microscopy and confocal laser scanning microscopy confirmed extensive proliferation and migration of human bone marrow stromal cells on all surfaces evaluated. CONCLUSIONS: LASER-modified Ti surfaces modify the behaviour of hSSCs.(1) In particular, SSC(1) adhesion, osteogenic gene expression, cell morphology and cytoskeleton structure were affected. The current studies show Ti LASER modification can enhance the osseointegration between Ti and skeletal cells, with important implications for orthopaedic application.

Avaliação de implantes modificados por feixe de laser Nd: YAG e implantes de superfície usinada empregando-se MEV e torque reverso / Evaluation of implants modified by laser beam Nd: YAG and machined surface implants employing SEM and reverse torque test

Implantes de Ti-cp foram irradiados utilizando-se feixe de laser Nd:YAG para serem comparados com implantes de superfície usinada. As amostras foram inseridas cirurgicamente na tíbia de coelhos. Após o período de oito semanas de cicatrização os animais foram sacrificados e os implantes retirados empregando-se teste de torque reverso. A análise por MEV-EDS da superfície irradiada com laser apresentou maior formação óssea em relação à superfície usinada. Ao teste de torque reverso o implante irradiado laser apresentou o valor de torque médio de 40 Ncm e o implante usinado 24 Ncm. Os resultados obtidos indicaram que a superfície irradiada por feixe de laser apresentou maior interação e adesão de tecido ósseo.

Ti-cp implants were irradiated using laser beam Nd:YAG to be compared with machined surface implants. The samples were inserted into rabbit tibia. After an eight-week healing period, the animals were sacrificed and the implants removed under reverse torque test. SEM-EDS analysis of the laser-irradiated surface showed higher bone formation than at the machined surface. The reverse torque test of the laser-irradiated implants showed medium torque values of 40 Ncm and machined implants of 24 Ncm. The results showed that the use of laser treatment imparts higher bone tissue interaction and adhesion.