Citrate zinc hydroxyapatite nanorods with enhanced cytocompatibility and osteogenesis for bone regeneration.

The development of biomaterials that mimicking the hydroxyapatite nanoparticles existent in the immature bone tissue is crucial, especially to accelerate the bone remodeling and regeneration. In this work, it was developed for the first time, hydroxyapatite nanoparticles (NPs) incorporating citrate and zinc (cit-Zn-Hap) in their composition towards a one-step hydrothermal procedure. For comparison purposes, hydroxyapatite NPs incorporating only zinc (Zn-Hap) or citrate (cit-Hap), as well as hydroxyapatite without any of these elements (Hap) were synthesised. The physicochemical characterization was carried out reveling that, the presence of zinc on hydroxyapatite (cit-Zn-Hap), reduced the size of nanoparticles, changed the phosphate environment and decreased the surface charge when compared with cit-Hap nanoparticles. The osteogenic potential of cit-Zn-Hap NPs was analysed in human bone marrow-derived stromal cells (BMSCs), in the absence of osteoinductive factors. NPs were internalized by endocytosis appearing trapped in endosomes and lysosomes scattered through the cytoplasm. Exposure to these NPs resulted in a significant induction of ALP activity, extracellular matrix mineralization, and gene expression of early and later osteogenic transcription factors, as well as of osteoblastic markers. The osteoinductive effect might be regulated, at least in part, by the increased signalling through the canonical WNT pathway. Evaluation of the cell behaviour following exposure to Zn-Hap and cit-Hap strongly suggested a synergistic effect of citrate and Zn in cit-Zn-Hap NPs towards the induction of the osteogenic commitment and functionality of BMSCs. These findings will allow the design of new biomimetic hydroxyapatite nanoparticles with great potential for bone regeneration.

Surface and mechanical properties of a nanostructured citrate hydroxyapatite coating on pure titanium.

The presence of a biomimetic HAP coating on titanium surface, which reduces the structural stiffness, is essential to improve implants biocompatibility and osteointegration. In this study, new citrate-HAP (cHAP) coatings were produced by a simple hydrothermal method on pure titanium (Ti) surface, without requiring any additional pretreatment on this metal surface. The formed cHAP coatings consisting of nanorod-like hydroxyapatite particles, conferred nanoroughness and wettability able to endow improved biological responses. Indeed, the presence of citrate species in the precipitate medium seems to be responsible for controlling the morphology of the new coatings. The presence of citrate groups on the surface of cHAP coatings, identified by chemical composition analysis, due to their implication in bone metabolism can additionally bring an add-value for bone implant applications. From a mechanical point of view, the Finite Element algorithm showing that cHAP coatings tend to decrease the mechanical stress at pure Ti, further favors these new coatings applicability. Overall, the simple and expedite strategy used to developed new biomimetic coatings of citrate-HAP resulted in improved physicochemical, morphological and mechanical properties of Ti, which can endeavor improved implantable materials in bone healing surgical procedures.