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Krzysztof Marycz,1,2 Agnieszka Smieszek,1 Klaudia Marcinkowska,1 Mateusz Sikora,1 Eliza Turlej,1 Paulina Sobierajska,3 Adrian Patej,3 Alina Bienko,4 Rafal J Wiglusz3 1The Department of Experimental Biology, The Faculty of Biology and Animal Science, University of Environmental and Life Sciences, Wroclaw, Poland; 2International Institute of Translational Medicine, Malin, Poland; 3Institute of Low Temperature and Structure Research, PAS, Wroclaw, Poland; 4Faculty of Chemistry, University of Wroclaw, Wroclaw, PolandCorrespondence: Krzysztof MaryczThe Department of Experimental Biology, The Faculty of Biology and Animal Science, University of Environmental and Life Sciences Wroclaw, Norwida 27B St, Wroclaw, 50– 375, PolandTel +48 71 320 5201Email krzysztof.marycz@upwr.edu.plPurpose: Osteoporosis results in a severe decrease in the life quality of many people worldwide. The latest data shows that the number of osteoporotic fractures is becoming an increasing international health service problem. Therefore, a new kind of controllable treatment methods for osteoporotic fractures is extensively desired. For that reason, we have manufactured and evaluated nanohydroxyapatite (nHAp)-based composite co-doped with iron oxide (IO) nanoparticles. The biomaterial was used as a matrix for the controlled delivery of miR-21-5p and miR-124-3p, which have a proven impact on bone cell metabolism.Methods: The nanocomposite Ca5(PO4)3OH/Fe3O4 (later called nHAp/IO) was obtained by the wet chemistry method and functionalised with microRNAs (nHAp/IO@miR-21/124). Its physicochemical characterization was performed using XRPD, FT-IR, SEM-EDS and HRTEM and SAED methods. The modulatory effect of the composite was tested in vitro using murine pre-osteoblasts MC3T3-E1 and pre-osteoclasts 4B12. Moreover, the anti-inflammatory effects of biomaterial were analysed using a model of LPS-treated murine macrophages RAW 264.7. We have analysed the cells’ viability, mitochondria membrane potential and oxidative stress under magnetic field (MF+) and without (MF-). Moreover, the results were supplemented with RT-qPCR and Western blot assays to evaluate the expression profile for master regulators of bone metabolism.Results: The results indicated pro-osteogenic effects of nHAp/IO@miR-21/124 composite enhanced by exposure to MF. The enhanced osteogenesis guided by nHAp/IO@miR-21/124 presence was associated with increased metabolism of progenitor cells and activation of osteogenic markers (Runx-2, Opn, Coll-1). Simultaneously, nanocomposite decreased metabolism and differentiation of pre-osteoclastic 4B12 cells accompanied by reduced expression of CaII and Ctsk. Obtained composite regulated viability of bone progenitor cells and showed immunomodulatory properties inhibiting the expression of inflammatory markers, ie, TNF-α, iNOs or IL-1β, in LPS-stimulated RAW 264.7 cells.Conclusion: We have described for the first time a new concept of osteoporosis treatment based on nHAp/IO@miR-21/124 application. Obtained results indicated that fabricated nanocomposite might impact proper regeneration of osteoporotic bone, restoring the balance between osteoblasts and osteoclast.Keywords: nanocomposites, hydroxyapatite, osteoblasts, osteoclasts, osteoporosis

Purpose: The presented study aimed to investigate the effects of Fe 3O 4 nanoparticles and static magnetic field on osteoblast and osteoclasts’ metabolic activity. Methods: Magnetic nanoparticles were prepared by a wet chemical co-precipitation process and analyzed using X-ray powder diffraction, high-resolution transmission electron microscope (HRTEM), dynamic light scattering (DLS), laser Doppler velocimetry, Raman and the Mossbauer spectroscopy. In vitro experiments were performed using MC3T3, 4B12 and RAW 264.7 cell lines. Cells were cultured in the presence of nanoparticles and with or without exposure to the magnetic field. Proteins were investigated with Western blotting and immunofluorescence and Western blot. Gene expression was analyzed with a quantitative real-time polymerase chain reaction. Results: Obtained particles were in the nano-range (average size around 50 nm) and had a spherical-like morphology. The typical hydrodynamic size was in the range 178– 202 nm and Zeta potential equaled – 9.51 mV. Mossbauer spectrum corresponds to the Fe+3 ions in tetrahedral (A) and Fe+3 and Fe+2 ions in octahedral (B) sites of Fe 3O 4. In vitro study revealed cytocompatibility and anti-inflammatory effects of fabricated nanoparticles. Furthermore, it was shown that nanoparticles combined with magnetic field exposure enhance osteogenic differentiation of MC3T3 cells by upregulation of RUNX-2 activity. Under the same experimental condition, nanoparticles and magnetic field decreased osteoclastogenesis of 4B12 by the induction of apoptosis through the mitochondrial-dependent pathway. Conclusion: Fe 3O 4 nanoparticles together with magnetic field can be applied for the fabrication of novel biomaterials for the treatment of bone disorders related to bone loss in which a balance between bone-forming and resorbing cells is disturbe

BackgroundPrevalence of osteoporosis is rapidly growing and so searching for novel therapeutics. Yet, there is no drug on the market available to modulate osteoclasts and osteoblasts activity simultaneously. Thus in presented research we decided to fabricate nanocomposite able to: (i) enhance osteogenic differentiation of osteoblast, (i) reduce osteoclasts activity and (iii) reduce pro-inflammatory microenvironment. As a consequence we expect that fabricated material will be able to inhibit bone loss during osteoporosis.ResultsThe α-Fe2O3/γ-Fe2O3nanocomposite (IOs) was prepared using the modified sol–gel method. The structural properties, size, morphology and Zeta-potential of the particles were studied by means of XRPD (X-ray powder diffraction), SEM (Scanning Electron Microscopy), PALS and DLS techniques. The identification of both phases was checked by the use of Raman spectroscopy and Mössbauer measurement. Moreover, the magnetic properties of the obtained IOs nanoparticles were determined. Then biological properties of material were investigated with osteoblast (MC3T3), osteoclasts (4B12) and macrophages (RAW 264.7) in the presence or absence of magnetic field, using confocal microscope, RT-qPCR, western blot and cell analyser. Here we have found that fabricated IOs: (i) do not elicit immune response; (ii) reduce inflammation; (iii) enhance osteogenic differentiation of osteoblasts; (iv) modulates integrin expression and (v) triggers apoptosis of osteoclasts.ConclusionFabricated by our group α-Fe2O3/γ-Fe2O3nanocomposite may become an justified and effective therapeutic intervention during osteoporosis treatment.

Due to its increased prevalence, osteoporosis (OP) represents a great challenge to health care systems and brings an economic burden. To overcome these issues, treatment plans that suit the need of patients should be developed. One of the approaches focuses on the fabrication of personalized biomaterials, which can restore the balance and homeostasis of disease-affected bone. In the presented study, we fabricated nanometer crystalline hydroxyapatite (nHAp) and iron oxide (IO) nanoparticles stabilized with APTES and investigated whether they can modulate bone cell metabolism and be useful in the fabrication of personalized materials for OP patients. Using a wide range of molecular techniques, we have shown that obtained nHAp@APTES promotes viability and RUNX-2 expression in osteoblasts, as well as reducing activity of critical proinflammatory cytokines while inhibiting osteoclast activity. Materials with APTES modified with nHAp incorporated with IO nanoparticles can be applied to support the healing of osteoporotic bone fractures as they enhance metabolic activity of osteoblasts and diminish osteoclasts’ metabolism and inflammation.

The main aim of our research was to investigate antiadhesive and antibiofilm properties of nanocrystalline apatites doped and co-doped with noble metal ions (Ag+, Au+, and Pd2+) against selected drug-resistant strains of Enterococcus faecalis and Staphylococcus aureus. The materials with the structure of apatite (hydroxyapatite, nHAp; hydroxy-chlor-apatites, OH-Cl-Ap) containing 1 mol% and 2 mol% of dopants and co-dopants were successfully obtained by the wet chemistry method. The majority of them contained an additional phase of metallic nanoparticles, in particular, AuNPs and PdNPs, which was confirmed by the XRPD, FTIR, UV–Vis, and SEM–EDS techniques. Extensive microbiological tests of the nanoapatites were carried out determining their MIC, MBC value, and FICI. The antiadhesive and antibiofilm properties of the tested nanoapatites were determined in detail with the use of fluorescence microscopy and computer image analysis. The results showed that almost all tested nanoapatites strongly inhibit adhesion and biofilm production of the tested bacterial strains. Biomaterials have not shown any significant cytotoxic effect on fibroblasts and even increased their survival when co-incubated with bacterial biofilms. Performed analyses confirmed that the nanoapatites doped and co-doped with noble metal ions are safe and excellent antiadhesive and antibiofilm biomaterials with potential use in the future in medical sectors.

The main aim of our research was to investigate antiadhesive and antibiofilm properties of nanocrystalline apatites doped and co-doped with noble metal ions (Ag

A new combination of Toceranib (Toc; 5-[(5Z)-(5-Fluoro-2-oxo-1,2-dihydro-3H-indol-3-ylidene)methyl]-2,4-dimethyl-N-[2-(pyrrolidin-1-yl)ethyl]-1H-pyrrole-3-carboxamide) with nanohydroxyapatite (nHAp) was proposed as an antineoplastic drug delivery system. Its physicochemical properties were determined as crystallinity, grain size, morphology, zeta potential and hydrodynamic diameter as well as Toceranib release. The crystalline nanorods of nHAp were synthesised by the co-precipitation method, while the amorphous Toceranib was obtained by its conversion from the crystalline form during nHAp–Toc preparation. The surface interaction between both compounds was confirmed using Fourier-transform infrared spectroscopy (FT-IR), ultraviolet-visible spectroscopy (UV–Vis) and scanning electron microscopy with energy-dispersive X-ray spectroscopy (SEM-EDS). The nHAp–Toc showed a slower and prolonged release of Toceranib. The release behaviour was affected by hydrodynamic size, surface interaction and the medium used (pH). The effectiveness of the proposed platform was tested by comparing the cytotoxicity of the drug combined with nHAp against the drug itself. The compounds were tested on NI-1 mastocytoma cells using the Alamar blue colorimetric technique. The obtained results suggest that the proposed platform shows high efficiency (the calculated IC50 is 4.29 nM), while maintaining the specificity of the drug alone. Performed analyses confirmed that nanohydroxyapatite is a prospective drug carrier and, when Toceranib-loaded, may be an idea worth developing with further research into therapeutic application in the treatment of canine mast cell tumour.

Li+/Eu3+ dual-doped calcium apatite analogues were fabricated using a microwave stimulated hydrothermal technique. XRPD, FT-IR, micro-Raman spectroscopy, TEM and SAED measurements indicated that obtained apatites are single-phased, crystallize with a hexagonal structure, have similar morphology and nanometric size as well as show red luminescence. Lithium effectively modifies the local symmetry of optical active sites and, thus, affects the emission efficiency. Moreover, the hydrodynamic size and surface charge of the nanoparticles have been extensively studied. The protein adsorption (lysozyme, LSZ, bovine serum albumin, BSA) on the nanoparticle surface depended on the type of cationic dopant (Li+, Eu3+) and anionic group (OH−, Cl−, F−) of the apatite matrix. Interaction with LSZ resulted in a positive zeta potential, and the nanoparticles had the lowest hydrodynamic size in this protein medium. The cytotoxicity assessment was carried out on the human osteosarcoma cell line (U2OS), murine macrophages (J774.E), as well as human red blood cells (RBCs). The studied apatites were not cytotoxic to RBCs and J774.E cells, however, at higher concentrations of nanoparticles, cytotoxicity was observed against the U2OS cell line. No antimicrobial activity was detected against Gram-negative bacteria with one exception for P. aeruginosa treated with Li+-doped fluorapatite.

In the present work, nanohydroxyapatites (nHAp) doped with copper and/or zinc ions were investigated for the assessment of its antibacterial activity and biocompatibility. Three forms of material with diverse surfaces were tested: nanopowder in colloidal suspension, galactose hydrogel (3,6-Anhydro-α-l-Galacto-β-d-Galactan) scaffold and pellet. The structural and morphological properties of the obtained biomaterials were comprehensively determined by using: XRPD, FT-IR, SEM-EDS, AAS, XPS and EPR techniques. The antimicrobial active ions, mostly Cu2+, were successfully released from the apatite structure despite the material being suspended in the porous galactose hydrogel matrix. The colloidal solutions of nanohydroxyapatites on bacterial viability revealed moderate activity of Cu2+-doped materials against Escherichia coli strain and significant activity against Pseudomonas aeruginosa strain. The comparative study of bacterial attachment to the hydrogel and pellet surface indicated that hydrogels were more prone to be colonized by both tested strains. Moreover, an additive of the Cu2+ ion modified bacterial attachment and biofilms forming on nHAp:Cu2+ and nHAp:Cu2+-Zn2+ materials. In the case of hydrogels, the biofilms were scattered while these forming on other materials were more clumped. The cytotoxicity evaluation of tested biomaterials showed biocompatible properties of both nanomaterial colloidal solutions as well as galactose hydrogel eluates toward normal mouse osteoblast cell lines (7F2) and human chondrocytes (TC28A2) and osteosarcoma cell line (U2OS). The biocompatibility of tested materials was additionally confirmed by conducting a hemolysis assay which showed full hemocompatibility of nanopowder colloidal solutions and galactose-based materials. Furthermore, unaltered red blood cell morphology was visible after a short and long time of incubation with the obtained biomaterials by using confocal laser scanning microscopy (CLSM). The comparison research provided data of 7F2, TC28 and U2OS cell attachment to the galactose hydrogel surface.

The administration of three-in-one parenteral nutrition (PN) admixtures to pediatric patients requires special consideration, specifically concerning quality and physicochemical stability. The introduction of a new parenteral amino acid solution into the market prompted us to evaluate Aminoplasmal Paed-based PN admixtures’ stability. The study aimed to determine the physicochemical parameters of the chosen variations of PN admixtures and search for a correlation between its composition and those parameters. One hundred and sixty-eight variations of PN admixtures intended for patients weighing from 10 to 25 kg and aged from 1 to 12 years and differing in the quantitative composition of electrolytes were selected for the study. The samples were prepared using each of the four intravenous lipid emulsions dedicated to pediatric patients: Intralipid 20%, Clinoleic 20%, Lipidem 20%, and Smoflipid 20%. The stability of the PN admixtures was assessed by visual inspection and determination of pH, osmolality, zeta potential, and hydrodynamic mean droplet diameter (MDD) immediately upon preparation and after seven days of storage at the temperature of 5 ± 1 °C with light protection. Pearson’s correlation was used to quantify the relationships between selected ingredients of the PN admixtures and the physicochemical parameters. The PN admixtures were characterized by pH ranging from 5.91 to 7.04, osmolality ranging from 1238 to 1678 mOsm/kg, and zeta potential ranging from −41.3 to −2.16 mV. The changes in pH and osmolality after seven days of storage did not exceed 0.2 and 4.4%, respectively. The homogeneity of the PN admixtures was confirmed by determining the polydispersity index, which ranged from 0.06 to 0.2. The MDD of the studied formulas ranged from 235 to 395 nm and from 233 to 365 nm immediately upon preparation and after the storage period, respectively. Correlations between selected components of the PN admixtures and some physicochemical parameters were found. All Aminoplasmal Paed 10%-based PN admixtures were characterized by appropriate physicochemical quality to be administered via the central veins, both immediately upon preparation and after seven days of storage at the temperature of 5 ± 1 °C with light protection. The applied electrolyte concentrations ranges and types of lipid emulsions in the selected macronutrient quantitative compositions allowed the PN admixtures to remain stable for seven days within the specified limits.

Lithium (Li+) ion due to its excellent bioactivity is one of the most well-studied element in bone-tissue engineering. In this study, we fabricated nanohydroxyapatite (nHAp) doped with Li+ ions (5 mol% Li+:nHAp) and co-doped with lanthanide ions. We investigated the effects of nHAp, 5 mol% Li+:nHAp or Li+ alone, on osteogenic differentiation of human Adipose Tissue-derived Stem Cells (hASCs), their proliferation, mitochondrial dynamics and apoptosis. Moreover, we monitored cell proliferation after treatment with samarium (III) (Sm3+) and europium (III) (Eu3+) ions co-doped 5 mol% Li+:nHAp as well as their luminescent property. The hASCs treated with 5 mol% Li+:nHAp and Li+ ions proliferated more rapidly and differentiated effectively than control cells without undergoing apoptosis. Both, 5 mol% Li+:nHAp and Li+ ions improved osteogenic differentiation of hASCs. Moreover they decreased expression of glycogen synthase kinase 3β (GSK3β) while increased β-catenin mRNA level. In addition, Li+, nHAp and 5 mol% Li+:nHAp improved mitochondrial dynamics and enhanced expression of neural differentiation marker genes. Collectively, the study indicates on pro-osteogenic and anti-apoptotic properties of nHAp doped with Li+ and Li+ alone. Moreover, unique properties of 5 mol% Li+:nHAp and 5 mol% Li+:nHAp co-doped with rare earth ions, such as Sm3+ and Eu3+ have shed a promising light on their potential application in theranostics.

In response to the need of new materials characterized by the spectroscopic properties in the near-infrared window in biological tissues (700–1100 nm), a new type of fluorapatites co-doped with Yb3+ and Tb3+ ions has been synthesized. The nanostructured fluorapatites were prepared using the so-called co-precipitation method and heat-treated in a broad temperature range (600–1100 °C). The structural and morphological properties of the obtained materials were determined by using XRPD (X-ray powder diffraction), TEM (transmission electron microscopy) and SEM (scanning electron microscopy) techniques. The crystallites size was verified and calculated by applying the Rietveld refinement. The primary size of the particles was estimated by TEM analysis and was found to be in the range from 30 nm × 15 nm for the sample annealed at 700 °C to 120 nm × 90 nm for the sample annealed at 1000 °C. Moreover, the formation of defects involving the doping process and their contribution to the charge compensation mechanism has also been discussed. Furthermore, the spectroscopic properties of the obtained materials have been investigated in detail with emission spectra, power dependence relations and luminescence kinetics. The co-doped fluorapatites demonstrated not only Stokes, but also anti-Stokes green luminescence (up-conversion (UC)) depending on the laser power. It was also found that there was weak emission from 5D3 energy level under UV irradiation, not detected under NIR excitation. The cooperative energy transfer (CET) was suggested as the main process responsible for the UC luminescence.

In the present study, nanocrystalline Ca–Sr hydroxyapatites structurally modified with Li+ ions as well as co-doped with Eu3+ ions were prepared by the hydrothermal method assisted by heat treatment. The structural and morphological properties of the studied materials were determined by using XRPD (X-ray powder diffraction), ATR-FTIR (attenuated total reflectance-FTIR spectroscopy) and SEM (scanning electron microscopy) techniques. According to Rietveld analysis, the crystallite sizes were calculated to be in the range of 48–67 nm and were close to the value obtained by SEM. Based on X-ray diffraction and IR absorption, the structural changes after the substitution of divalent calcium ions by other cations in the apatite crystal lattice were identified. The changes appeared as a shift of the (0 0 2) plane position as well as a shift of the PO43− and OH− bands. In this case, a possible charge compensation mechanism was investigated based on the Kroger–Vink notation. The investigation of the luminescence properties (emission, emission excitation and emission kinetics) of the studied materials as a function of Li+ ion concentrations was carried out. The luminescence behavior of Eu3+ ions in the samples was analyzed by the Judd–Ofelt (J–O) theory, and the J–O parameters were estimated and discussed.

Hydroxyapatite has been used in medicine for many years as a biomaterial or a cover for other biomaterials in orthopedics and dentistry. This study characterized the physicochemical properties (structure, particle size and morphology, surface properties) of Li+- and Li+/Eu3+-doped nanohydroxyapatite obtained using the wet chemistry method. The potential regenerative properties against neurite damage in cultures of neuron-like cells (SH-SY5Y and PC12 after differentiation) were also studied. The effect of nanohydroxyapatite (nHAp) on the induction of repair processes in cell cultures was assessed in tests of metabolic activity, the level of free oxygen radicals and nitric oxide, and the average length of neurites. The study showed that nanohydroxyapatite influences the increase in mitochondrial activity, which is correlated with the increase in the length of neurites. It has been shown that the doping of nanohydroxyapatite with Eu3+ ions enhances the antioxidant properties of the tested nanohydroxyapatite. These basic studies indicate its potential application in the treatment of neurite damage. These studies should be continued in primary neuronal cultures and then with in vivo models.

New fluconazole-loaded, 6-anhydro-α-L-galacto-β-D-galactan hydrogels incorporated with nanohydroxyapatite were prepared and their physicochemical features (XRD, X-ray Diffraction, SEM-EDS, Scanning Electron Microscopy-Energy Dispersive X-ray Spectroscopy, ATR-FTIR, Attenuated Total Reflectance-Fourier Transform Infrared Spectroscopy), fluconazole release profiles and enzymatic degradation were determined. Antifungal activity of pure fluconazole was tested using Candida species (C. albicans, C. tropicalis, C. glabarata), Cryptococcus species (C. neoformans, C. gatti) and Rhodotorula species (R. mucilaginosa, R. rubra) reference strains and clinical isolates. Standard microdilution method was applied, and fluconazole concentrations of 2–250 µg/mL were tested. Moreover, biofilm production ability of tested isolates was tested on the polystyrene surface at 28 and 37 ± 0.5 °C and measured after crystal violet staining. Strains with the highest biofilm production ability were chosen for further analysis. Confocal microscopy photographs were taken after live/dead staining of fungal suspensions incubated with tested hydrogels (with and without fluconazole). Performed analyses confirmed that polymeric hydrogels are excellent drug carriers and, when fluconazole-loaded, they may be applied as the prevention of chronic wounds fungal infection.

Krzysztof Marycz,1,2 Agnieszka Śmieszek,1 Katarzyna Kornicka-Garbowska,1,2 Ariadna Pielok,1 Maciej Janeczek,3 Anna Lipińska,3 Anna Nikodem,4 Jarosław Filipiak,4 Paulina Sobierajska,5 Jean-Marie Nedelec,6 Rafał J Wiglusz2,5 1Department of Experimental Biology, Wroclaw University of Environmental and Life Sciences, Wroclaw, 50-375, Poland; 2International Institute of Translational Medicine, Malin, 55-124, Poland; 3Department of Biostructure and Animal Physiology, Wroclaw University of and Life Sciences, Wroclaw, 51-631, Poland; 4Department of Mechanics, Materials and Biomedical Engineering, Wroclaw University of Science and Technology, Wybrzeze Wyspianskiego 27, 50-370 Wrocław, Poland; 5Institute of Low Temperature and Structure Research, Polish Academy of Sciences, Okolna 2, 50-422 Wroclaw, Poland; 6Universite Clermont Auvergne, Clermont Auvergne INP, CNRS, ICCF, Clermont-Ferrand, FranceCorrespondence: Krzysztof MaryczThe Department of Experimental Biology, The Faculty of Biology and Animal Science, University of Environmental and Life Sciences Wroclaw, Norwida 27B St, Wroclaw, 50– 375, PolandTel +48 71 320 5201Email krzysztof.marycz@upwr.edu.plPurpose: Healing of osteoporotic defects is challenging and requires innovative approaches to elicit molecular mechanisms promoting osteoblasts-osteoclasts coupling and bone homeostasis.Methods: Cytocompatibility and biocompatibility of previously characterised nanocomposites, i.e Ca5(PO4)3OH/Fe3O4 (later called nHAp/IO) functionalised with microRNAs (nHAp/IO@miR-21/124) was tested. In vitro studies were performed using a direct co-culture system of MC3T3-E1 pre-osteoblast and 4B12 pre-osteoclasts. The analysis included determination of nanocomposite influence on cultures morphology (confocal imaging), viability and metabolic activity (Alamar Blue assay). Pro-osteogenic signals were identified at mRNA, miRNA and protein level with RT-qPCR, Western blotting and immunocytochemistry. Biocompatibility of biomaterials was tested using bilateral cranial defect performed on a senescence-accelerated mouse model, ie SAM/P6 and Balb/c. The effect of biomaterial on the process of bone healing was monitored using microcomputed tomography.Results: The nanocomposites promoted survival and metabolism of bone cells, as well as enhanced functional differentiation of pre-osteoblasts MC3T3-E1 in co-cultures with pre-osteoclasts. Differentiation of MC3T3-E1 driven by nHAp/IO@miR-21/124 nanocomposite was manifested by improved extracellular matrix differentiation and up-regulation of pro-osteogenic transcripts, ie late osteogenesis markers. The nanocomposite triggered bone healing in a cranial defect model in SAM/P6 mice and was replaced by functional bone in Balb/c mice.Conclusion: This study demonstrates that the novel nanocomposite nHAp/IO can serve as a platform for therapeutic miRNA delivery. Obtained nanocomposite elicit pro-osteogenic signals, decreasing osteoclasts differentiation, simultaneously improving osteoblasts metabolism and their transition toward pre-osteocytes and bone mineralisation. The proposed scaffold can be an effective interface for in situ regeneration of osteoporotic bone, especially in elderly patients.Keywords: senile osteoporosis, osteoblasts, osteoclasts, nanohydroxyapatite, iron oxide nanoparticles, small non-coding RNA

The research has been carried out with a focus on the assessment of the antimicrobial efficacy of pure nanohydroxyapatite, Cu2+-doped nanohydroxyapatite, ozonated olive oil-loaded nanohydroxyapatite, and Cu2+-doped nanohydroxyapatite, respectively. Their potential antimicrobial activity was investigated against Streptococcus mutans, Lactobacillus rhamnosus, and Candida albicans. Among all tested materials, the highest efficacy was observed in terms of ozonated olive oil. The studies were performed using an Ultraviolet&ndash, Visible spectrophotometry (UV-Vis), electron microscopy, and statistical methods, by determining the value of Colony-Forming Units (CFU/mL) and Minimal Inhibitory Concentration (MIC).

In the present study, a nanoapatite-mediated delivery system for imatinib has been proposed. Nanohydroxyapatite (nHAp) was obtained by co-precipitation method, and its physicochemical properties in combination with imatinib (IM) were studied by means of XRPD (X-ray Powder Diffraction), SEM-EDS (Scanning Electron Microscopy-Energy Dispersive X-ray Spectroscopy), FT-IR (Fourier-Transform Infrared Spectroscopy), absorption spectroscopy as well as DLS (Dynamic Light Scattering) techniques. The obtained hydroxyapatite was defined as nanosized rod-shaped particles with high crystallinity. The amorphous imatinib was obtained by conversion of its crystalline form. The beneficial effects of amorphous pharmaceutical agents have been manifested in the higher dissolution rate in body fluids improving their bioavailability. Imatinib-to-hydroxyapatite interactions on the surface were confirmed by SEM images as well as absorption and FT-IR spectroscopy. The cytotoxicity of the system was tested on NI-1, L929, and D17 cell lines. The effectiveness of imatinib was not affected by nHAp modification. The calculated IC50 values for drug-modified nHAp were similar to those for the drug itself. However, higher cytotoxicity was observed at higher concentrations of imatinib, in comparison with the drug alone.

The presented study aimed to investigate the effects of FeMagnetic nanoparticles were prepared by a wet chemical co-precipitation process and analyzed using X-ray powder diffraction, high-resolution transmission electron microscope (HRTEM), dynamic light scattering (DLS), laser Doppler velocimetry, Raman and the Mössbauer spectroscopy. In vitro experiments were performed using MC3T3, 4B12 and RAW 264.7 cell lines. Cells were cultured in the presence of nanoparticles and with or without exposure to the magnetic field. Proteins were investigated with Western blotting and immunofluorescence and Western blot. Gene expression was analyzed with a quantitative real-time polymerase chain reaction.Obtained particles were in the nano-range (average size around 50 nm) and had a spherical-like morphology. The typical hydrodynamic size was in the range 178-202 nm and Zeta potential equaled -9.51 mV. Mössbauer spectrum corresponds to the FeFe

Calcium hydroxyapatite porous nanoceramics (ncHAP) were prepared in the form of granules utilizing raw, high purity HAP nanopowder and methylcellulose (MC) in ratio of 60/40 wt% with the final moulding at high temperature (800–1000°C). The ncHAP's were studied by means of XRD, TEM, SEM techniques for structural and morphological characterizations. BET method was used in order to estimate the surface area, pore diameter and volume as well as the average particle size. It was found that particle size of raw material and nanoceramic prior heating (with MC) was 33 nm whereas ncHAP's size has significantly increased with sintering temperature from 136 nm up to 955 nm. X-ray diffraction clearly showed that all materials were crystallized and under given process parameters the HAP underwent dehydratation event as the content of β-TCP (tricalcium phosphate) has increased along with temperature up to 24%. The presence of β-TCP in porous nanoceramic material, might be advantageous since both phosphates have different solubility in biological media thus it is possible to affect the resorption speed. The antimicrobial activity of raw materials and nanoceramics against Pseudomonas aeruginosa as a representative of the Gram-negative bacteria has been determined in vitro and in vivo on Galleria mellonella larvae model. The results confirm that definitely nanobased biomaterials of calcium hydroxyapatite are very promising compound with high potential in biomedical applications.

Spinal cord injuries (SCI) often require simultaneous regeneration of nerve tissue and bone. Hydroxyapatites are described as bioresorbable materials with proper biocompatibility and osteoconductivity, therefore its application for spinal surgery is considered. In this paper, we present repeatable method for developing nanocrystalline calcium hydroxyapatites structurally modified with Li+ ions (nHAP:Li+). Obtained biomaterials were profoundly characterized in terms of their physicochemical properties. Moreover, we have shown that nHAP:Li+ doped with europium (Eu3+) may serve as a theranostic agent, what additionally extend its potential usage for SCI treatment. The biocompatibility of nHAP:Li+ was determined using human olfactory ensheathing cells (hOECs) and adipose tissue-derived multipotent stromal cells (hASCs). Both population of cells are eagerly applied for cell-based therapies in SCI, mainly due to their paracrine activity. The extensive in vitro studies showed that nHAP:Li+ promotes the cells proliferation, viability and cell-cell interactions. Obtained results provides encouraging approach that may have potential application in regenerative medicine and that could fulfil the promise of personalized medicine – important in SCI treatment.

An assessment of biomaterial cytotoxicity is a prerequisite for evaluation of its clinical potential. A material is considered toxic while the cell viability decreases under 70% of the control. However, extracts of certain materials are likely to reduce the cell viability due to the intense ions adsorption from culture medium (e.g. highly bioactive ceramics of high surface area). Thus, the standard ISO 10993-5 procedure is inappropriate for cytotoxicity evaluation of ceramics of high specific surface area because biomaterial extract obtained in this method (ions-depleted medium) is not optimal for cell cultures per se. Therefore, a simple test was designed as an alternative to ISO 10993-5 standard for cytotoxicity evaluation of the biomaterials of high surface area and high ions absorption capacity. The method, presented in this paper, included the evaluation of ceramics extract prepared according to corrected procedure. The corrected extract was found not cytotoxic (cell viability above 70%), suggesting that modified method for cytotoxicity evaluation of ions-adsorbing ceramics is more appropriate than ISO 10993-5 standard. For such biomaterials, the term "false" cytotoxicity is more suitable. Moreover, it was noted that NRU assay and microscopic observations should be recommended for cytotoxicity evaluation of ceramics of high surface area.

Hexagonal nanocrystalline powders of the non-doped Ca10(PO4)6(OH)2 as well as activated with Ag+ and Eu3+ ions were synthesized by using different wet chemistry methods. Moreover, the obtained hydroxyapatite was loaded with Ag0, as well as nitroimidazole antimicrobials: metronidazole and tinidazole. The structural properties of the products were analyzed by X-ray diffraction (XRD), scanning (SEM) and transmission (TEM) electron microscopy as well as infrared (IR) and Raman spectroscopy. The photoluminescence properties of the Eu3+ and Ag+ co-doped Ca10(PO4)6(OH)2 were characterized via the PL emission, excitation spectra and the luminescence decay curve. The antimicrobial activity of the obtained materials against Prevotella bivia and Parabacteroides distasonis was studied. The cytotoxicity assessment was carried out on the human osteosarcoma cell line (U2OS) as well as human red blood cells (RBC). The choice of the in vitro model was based on the fact that U2OS is a cancer cell line derived from bone tissue which is rich in apatites that play a pivotal role in the extracellular matrix formation. RBCs are the most abundant blood cells and they are used as a cell model in the study of biocompatibility of new prepared biocompounds with potential medical applications. The obtained multifunctional materials do not exhibit the haemolytic activity, therefore, they could be used as a promising antimicrobial agent and for anaerobic bacteria.

International audience; The nano-biphasic calcium phosphate co-doped with 1 mol% Er3+ and 5 mol% Yb3+ ions was prepared using Pechini's technique. The preparation of the biphasic calcium phosphate showed the removal of fluorine ions from the matrix. The structural properties and morphology of the particles were studied by means of XRD (X-ray powder diffraction), TEM (Transmission Electron Microscopy), and SEM (Scanning Electron Microscopy) techniques. The spectroscopic properties of Ca10(PO4)6F2/Ca3(PO4)2:Er3+,Yb3+ nano-biphasic calcium phosphate were investigated in detail using emission spectra, power dependence and emission kinetics. This material demonstrated intense green up-conversion depending on laser power. Additionally, the luminescence lifetimes for Er3+ ions were calculated from the measured decays. Power dependence of the green and red emissions was found to result from a temperature increase of nano-biphasic calcium phosphate that opens potential applications in theranostics.

Eu3+ doped fluorapatite nanopowders were synthesized by a microwave assisted hydrothermal technique and annealed in the 500–1000 °C temperature range for 3 h. The europium ion concentration was determined to be in the 0.5–10 mol% concentration range. The morphology and structural properties of Ca(10−3x)Eu2x(PO4)6F2 were determined by using TEM (transmission electron microscopy) and XRPD (X-ray powder diffraction) techniques, as well as the IR (infrared) and Raman spectroscopy. The average nanoparticle sizes were calculated by Rietveld refinement to be in the range from 30 nm for the as-prepared samples to above 100 nm for the materials thermally treated at 1000 °C. The particles developed into rectangular prism nanorods. The spectroscopic properties of the obtained materials were observed to be dependent on the dopant concentration and annealing temperature and were studied in detail. The Judd–Ofelt theory was applied and correlated with structural and optical properties to get detailed insight into the fluorapatite structures.

In response to the demand for new multifunctional materials characterized by high biocompatibility, hydrogel (HG) nanocomposites as a platform for bioactive compound delivery have been developed and fabricated. A specific crosslinking/copolymerization chemistry was used to construct hydrogels with a controlled network organization. The hydrogels were prepared using 3,6-anhydro-&alpha, l-galacto-&beta, d-galactan (galactose hydrogel) together with resveratrol (trans-3,5,4&prime, trihydroxystilbene) and calcium hydroxyapatite nanoparticles. The resveratrol was introduced in three different concentrations of 0.1, 0.5, and 1 mM. Nanosized calcium hydroxyapatite was synthesized by a microwave-assisted hydrothermal technique, annealed at 500 &deg, C for 3 h, and introduced at a concentration 10% (m/v). The morphology and structural properties of Ca10(PO4)6(OH)2 and its composite were determined by using XRPD (X-ray powder diffraction) techniques, as well as the absorption and IR (infrared) spectroscopy. The average nanoparticle size was 35 nm. The water affinity, morphology, organic compound release profile, and cytocompatibility of the obtained materials were studied in detail. The designed hydrogels were shown to be materials of biological relevance and of great pharmacological potential as carriers for bioactive compound delivery. Their cytocompatibility was tested using a model of human multipotent stromal cells isolated from adipose tissue (hASCs). The biomaterials increased the proliferative activity and viability of hASCs, as well as reduced markers of oxidative stress. In light of the obtained results, it has been thought that the designed materials meet the requirements of the tissue engineering triad, and may find application in regenerative medicine, especially for personalized therapies.

Spinal cord injuries (SCI) often require simultaneous regeneration of nerve tissue and bone. Hydroxyapatites are described as bioresorbable materials with proper biocompatibility and osteoconductivity, therefore its application for spinal surgery is considered. In this paper, we present repeatable method for developing nanocrystalline calcium hydroxyapatites structurally modified with Li

Osteoconductive drug delivery system composed of nanocrystalline calcium phosphates (Ca10(PO4)6(OH)2/β-Ca3(PO4)2) co-doped with Yb(3+)/Er(3+) ions loaded with Tetracycline antibiotic (TC) was developed. Their effect on human adipose derived mesenchymal stromal stem cells (hASCs) as a potential reconstructive biomaterial for bone tissue regeneration was studied. The XRD and TEM measurements were used in order to determine the crystal structure and morphology of the final products. The characteristics of nanocomposites with the TC and hASCs as potential regenerative materials as well as the antimicrobial activity of the nanoparticles against: Staphylococcus aureus ATCC 25923 as a model of the Gram-positive bacteria, Escherichia coli ATCC 8739 of the Gram-negative bacteria, were shown. These combinations can be a promising material for theranostic due to its regenerative, antimicrobial and fluorescent properties.

International audience; Eu3+ and Li+ ion co-doped fluorapatite (Ca10(PO4)6F2) nanocrystals were fabricated using a microwave stimulated hydrothermal technique followed by heat treatment at 500 °C. The concentration of Eu3+ ions was set to be 1 mol% and that of the Li+ ions was in the range of 0.5–5 mol% to investigate the site occupancy preference and charge compensation co-doping. The structural and morphological properties of the obtained samples were determined by using XRD (X-ray powder diffraction) and TEM (transmission electron microscopy) techniques as well as IR (infrared) and micro-Raman spectroscopy. The particle size was verified and calculated by the Rietveld method being in the range of 70–95 nm. The luminescence properties (the emission, excitation spectra and emission kinetics) of the Eu3+ ion-doped fluorapatite depending on the co-dopant (Li+ ions) were recorded. Significantly, fluorescence quenching by OH− groups was eliminated by F- ions and the luminescence intensity was enhanced by co-doping with Li+ ions. The simplified Judd–Ofelt (J–O) theory has been performed to explain a detailed analysis of the luminescence spectra.

Correction for ‘Effects of crystalline growth on structural and luminescence properties of Ca(10−3x)Eu2x(PO4)6F2 nanoparticles fabricated by using a microwave driven hydrothermal process’ by Katarzyna Zawisza et al., CrystEngComm, 2017, 19, 6936–6949.