Design, clinical applications and post-surgical assessment of bioresorbable 3D-printed craniofacial composite implants.

This study details the design, fabrication, clinical trials' evaluation, and analysis after the clinical application of 3D-printed bone reconstruction implants made of nHAp@PLDLLA [nanohydroxyapatite@poly(L-lactide-co-D,L-lactide)] biomaterial. The 3D-printed formulations have been tested as bone reconstruction Cranioimplants in 3 different medical cases, including frontal lobe, mandibular bone, and cleft palate reconstructions. Replacing one of the implants after 6 months provided a unique opportunity to evaluate the post-surgical implant obtained from a human patient. This allowed us to quantify physicochemical changes and develop a spatial map of osseointegration and material degradation kinetics as a function of specific locations. To the best of our knowledge, hydrolytic degradation and variability in the physicochemical and mechanical properties of the biomimetic, 3D-printed implants have not been quantified in the literature after permanent placement in the human body. Such analysis has revealed the constantly changing properties of the implant, which should be considered to optimize the design of patient-specific bone substitutes. Moreover, it has been proven that the obtained composition can produce biomimetic, bioresorbable and bone-forming alloplastic substitutes tailored to each patient, allowing for shorter surgery times and faster patient recovery than currently available methods.

New silicate-substituted hydroxyapatite materials doped with silver ions as potential antifungal agents.

BACKGROUND: Hydroxyapatites (HAp) are widely used as medical preparations for e.g., bone replacement or teeth implants. Incorporation of various substrates into HAp structures could enhance its biological properties, like biocompatibility or antimicrobial effects. Silver ions possess high antibacterial and antifungal activity and its application as HAp dopant might increase its clinical value. RESULTS: New silicate-substituted hydroxyapatites (HAp) doped with silver ions were synthesized via hydrothermal methods. The crystal structure of HAp was investigated by using the X-ray powder diffraction. Antifungal activity of silver ion-doped HAp (with 0.7 mol%, 1 mol% and 2 mol% of dopants) was tested against the yeast-like reference and clinical strains of Candida albicans, C. glabrata, C. tropicalis, Rhodotorula rubra, R. mucilaginosa, Cryptococcus neoformans and C. gattii. Spectrophotometric method was used to evaluate antifungal effect of HAp in SD medium. It was shown that already the lowest dopant (0.7 mol% of Ag+ ions) significantly reduced fungal growth at the concentration of 100 µg/mL. Increase in the dopant content and the concentration of HAp did not cause further growth inhibition. Moreover, there were some differences at the tolerance level to Ag+ ion-doped HAp among tested strains, suggesting strain-specific activity. CONCLUSIONS: Preformed studies confirm antimicrobial potential of hydroxyapatite doped with silver. New Ag+ ion-HAp material could be, after further studies, considered as medical agent with antifungal properties which lower the risk of a surgical-related infections.

The Study of Nanosized Silicate-Substituted Hydroxyapatites Co-Doped with Sr2+ and Zn2+ Ions Related to Their Influence on Biological Activities.

Nanosized silicate-substituted hydroxyapatites, characterized by the general formula Ca9.8-x-nSrnZnx(PO4)6-y(SiO4)y(OH)2 (where: n = 0.2 [mol%]; x = 0.5-3.5 [mol%]; y = 4-5 [mol%]), co-doped with Zn2+ and Sr2+ ions, were synthesized with the help of a microwave-assisted hydrothermal technique. The structural properties were determined using XRD (X-ray powder diffraction) and Fourier-transformed infrared spectroscopy (FT-IR). The morphology, size and shape of biomaterials were detected using scanning electron microscopy techniques (SEM). The reference strains of Klebsiella pneumoniae, Escherichia coli and Pseudomonas aeruginosa were used to assess bacterial survivability and the impact on biofilm formation in the presence of nanosilicate-substituted strontium-hydroxyapatites. Safety evaluation was also performed using the standard cytotoxicity test (MTT) and hemolysis assay. Moreover, the mutagenic potential of the materials was assessed (Ames test). The obtained results suggest the dose-dependent antibacterial activity of nanomaterials, especially observed for samples doped with 3.5 mol% Zn2+ ions. Moreover, the modification with five SiO4 groups enhanced the antibacterial effect; however, a rise in the toxicity was observed as well. No harmful activity was detected in the hemolysis assay as well as in the mutagenic assay (Ames test).

Investigation of Different Types of Micro- and Nanostructured Materials for Bone Grafting Application.

The insufficient volume of dental ridges is one of the most severe problems regarding an oral cavity. An inadequate amount can cause problems during various types of dental treatment. Its complexity originates from the etiopathogenesis of this problem. In this study, the representatives of auto-, allo-, and xenografts are compared. The physic-chemical differences between each of them were evaluated using XRD (X-ray Powder Diffraction), a SEM (Scanning Electron Microscopy), FT-IR (Fourier transformed infrared spectroscopy), and TGA (thermogravimetric analysis). Based on the SEM images, it was observed that the origin of the material has an influence on collagen fiber compact level and porosity. Following a comparison of FT-IR spectra and XRD, the crystal and chemical structures were described. Based on TGA, different water concentrations of the investigated materials, their high thermal stability, and concentration of inorganic phase, hydroxyapatite was determined. The presented study is important because it delivers information about chemical structure and its impact on bone regeneration. This knowledge should be taken into consideration by dental clinicians, because different types of bone grafts can accommodate the achievement of various goals.

Emission Enhancement and Energy Transfers in YV0.5P0.5O4 Nanoparticles Codoped with Eu3+ and Bi3+ Ions.

In this study, solid-state solutions of yttrium orthovanadate-phosphate with varying concentrations of codopants (Eu3+, Bi3+) have been obtained via coprecipitation. An ionic radii mismatch between V5+ and P5+ substituents is manifested in broad XRD lines. The sharpening of the XRD lines is observed with increasing bismuth ions concentration in the Eu3+ codoped YV0.5P0.5O4 matrix. The difference in the number of the Stark components for the 5D0 → 7FJ transitions indicates changes in the lattice and a number of possible Eu3+ sites. A thorough, systematic spectroscopic analysis of YV0.5P0.5O4: x mol % Eu3+, y mol % Bi3+ was conducted at room temperature and 5 K. Metal-to-metal energy transfers occurring between Eu3+, V5+, and Bi3+ optically active ions have been investigated. Additionally, efficiency of the Bi3+-Eu3+ energy transfer (ET) was calculated.

Investigation of topography effect on antibacterial properties and biocompatibility of nanohydroxyapatites activated with zinc and copper ions: In vitro study of colloids, hydrogel scaffolds and pellets.

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-ß-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.

Synergistic Effect of Toceranib and Nanohydroxyapatite as a Drug Delivery Platform-Physicochemical Properties and In Vitro Studies on Mastocytoma Cells.

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.

In Vitro Fracture Resistance of Endodontically Treated Premolar Teeth Restored with Prefabricated and Custom-Made Fibre-Reinforced Composite Posts.

(1) Background: The study aimed to compare and analyse the differences between the features of prefabricated fibre-reinforced composite (FRC) posts and custom-made FRC posts in the form of a tape and confirm the necessity of using FRC posts in teeth treated endodontically in comparison to direct reconstruction with a composite material. (2) Methods: Sixty premolars after endodontic treatment were used. The teeth were divided into four groups (n-15). Group 1: teeth with embedded prefabricated posts (Mirafit White); group 2: teeth with embedded prefabricated posts (Rebilda); group 3 teeth with embedded custom-made posts in the form of a tape (EverStick); group 4: teeth without a post restored with composite material. The compressive strength of the teeth was tested using the Instron-5944 testing machine until the sample broke. The crystal structure of the investigated posts was detected with the X-ray diffractometer (3) Results: During the experiment, the maximum values of forces at which the damage of the restored premolar teeth after endodontic treatment occurred were obtained. The best results were obtained for teeth rebuilt with Rebilda Posts (1119 N), while teeth with cemented Mirafit White posts were the weakest (968 N). Teeth without an embedded FRC post, rebuilt only with light-cured composite material, obtained the lowest value-859 N. (4) Conclusions: The use of FRC posts increases the resistance to damage of an endodontically treated tooth when compared to direct restoration with light-cured composite material.

The Influence of a Knitted Hydrophilic Prosthesis of Blood Vessels on the Activation of Coagulation System-In Vitro Study.

The replacement of affected blood vessels of the polymer material can cause imbalances in the blood haemostatic system. Changes in blood after the implantation of vascular grafts depend not only on the chemical composition but also on the degree of surface wettability. The Dallon® H unsealed hydrophilic knitted vascular prosthesis double velour was assessed at work and compare with hydrophobic vascular prosthesis Dallon®. Spectrophotometric studies were performed in the infrared and differential scanning calorimetry, which confirmed the effectiveness of the process of modifying vascular prostheses. Determination of the parameters of coagulation time of blood after contact in vitro with Dallon® H vascular prosthesis was also carried out. Prolongation of activated thromboplastin time, decreased activity of factor XII, IX and VIII, were observed. The prolonged thrombin and fibrinogen were reduced in the initial period of the experiment. The activity of plasminogen and antithrombin III and protein C were at the level of control value. The observed changes in the values of determined parameters blood coagulation do not exceed the range of referential values for those indexes. The observed changes are the result of considerable blood absorptiveness by the prosthesis of blood vessels and their sealing.

New Approach to Antifungal Activity of Fluconazole Incorporated into the Porous 6-Anhydro-α-l-Galacto-ß-d-Galactan Structures Modified with Nanohydroxyapatite for Chronic-Wound Treatments-In Vitro Evaluation.

New fluconazole-loaded, 6-Anhydro-α-l-Galacto-ß-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.

Quenching of the Eu3+ Luminescence by Cu2+ Ions in the Nanosized Hydroxyapatite Designed for Future Bio-Detection.

The hydroxyapatite nanopowders of the Eu3+-doped, Cu2+-doped, and Eu3+/Cu2+-co-doped Ca10(PO4)6(OH)2 were prepared by a microwave-assisted hydrothermal method. The structural and morphological properties of the products were investigated by X-ray powder diffraction (XRD), transmission electron microscopy techniques (TEM), and infrared spectroscopy (FT-IR). The average crystal size and the unit cell parameters were calculated by a Rietveld refinement tool. The absorption, emission excitation, emission, and luminescence decay time were recorded and studied in detail. The 5D0 → 7F2 transition is the most intense transition. The Eu3+ ions occupied two independent crystallographic sites in these materials exhibited in emission spectra: one Ca(1) site with C3 symmetry and one Ca(2) sites with Cs symmetry. The Eu3+ emission is strongly quenched by Cu2+ ions, and the luminescence decay time is much shorter in the case of Eu3+/Cu2+ co-doped materials than in Eu3+-doped materials. The luminescence quenching mechanism as well as the schematic energy level diagram showing the Eu3+ emission quenching mechanism using Cu2+ ions are proposed. The electron paramagnetic resonance (EPR) technique revealed the existence of at least two different coordination environments for copper(II) ion.

Investigation of Physicochemical Properties of the Structurally Modified Nanosized Silicate-Substituted Hydroxyapatite Co-Doped with Eu3+ and Sr2+ Ions.

In this paper, a series of structurally modified silicate-substituted apatite co-doped with Sr2+ and Eu3+ ions were synthesized by a microwave-assisted hydrothermal method. The concentration of Sr2+ ions was set at 2 mol% and Eu3+ ions were established in the range of 0.5-2 mol% in a molar ratio of calcium ion amount. The XRD (X-ray powder diffraction) technique and infrared (FT-IR) spectroscopy were used to characterize the obtained materials. The Kröger-Vink notation was used to explain the possible charge compensation mechanism. Moreover, the study of the spectroscopic properties (emission, emission excitation and emission kinetics) of the obtained materials as a function of optically active ions and annealing temperature was carried out. The luminescence behavior of Eu3+ ions in the apatite matrix was verified by the Judd-Ofelt (J-O) theory and discussed in detail. The temperature-dependent emission spectra were recorded for the representative materials. Furthermore, the International Commission on Illumination (CIE) chromaticity coordinates and correlated color temperature were determined by the obtained results.

Preparation of a New Biocomposite Designed for Cartilage Grafting with Antibiofilm Activity.

New polymer-inorganic composites with antibiofilm features based on the granulated poly(tetrafluoroethylene) (PTFE) and apatite materials were obtained using a standard hydraulic press. The study was performed in hydroxy- and fluorapatites doped with different amounts of silver ions and followed by heat treatment at 600 °C. The structural, morphological, and physicochemical properties were determined by X-ray powder diffraction (XRD), Fourier transform infrared (FT-IR) spectroscopy, scanning electron microscopy-energy-dispersive spectrometry (SEM-EDS), and transition electron microscopy (TEM). The antibacterial properties of the obtained materials were evaluated against Gram-negative pathogens such as Pseudomonas aeruginosa, Klebsiella pneumoniae, and Escherichia coli as well as against Gram-positive bacteria Staphylococcus epidermidis. The cytotoxicity assessment was carried out on the red blood cells (RBC) as a cell model for in vitro study. Moreover, the biofilm formation on the biocomposite surface was studied using confocal laser scanning microscopy (CLSM).

The Influence of Ozonated Olive Oil-Loaded and Copper-Doped Nanohydroxyapatites on Planktonic Forms of Microorganisms.

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-Visible spectrophotometry (UV-Vis), electron microscopy, and statistical methods, by determining the value of Colony-Forming Units (CFU/mL) and Minimal Inhibitory Concentration (MIC).

Three dimensional (3D) printed polylactic acid with nano-hydroxyapatite doped with europium(III) ions (nHAp/PLLA@Eu3+) composite for osteochondral defect regeneration and theranostics.

In the current research previously developed composites composed from poly (l-lactide) (PLLA) and nano-hydroxyapatite (10 wt% nHAp/PLLA) were functionalized with different concentrations of europium (III) (Eu3+). The aim of this study was to determine whether Eu3+ ions doped within the 10 wt% nHAp/PLLA scaffolds will improve the bioactivity of composites. Therefore, first set of experiments was designed to evaluate the effect of Eu3+ ions on morphology, viability, proliferation and metabolism of progenitor cells isolated from adipose tissue (hASC). Three different concentration were tested i.e. 1 mol%, 3 mol% and 5%mol. We identified the 10 wt% nHAp/PLLA@3 mol% Eu3+ scaffolds as the most cytocompatible. Further, we investigated the influence of the composites doped with 3 mol% Eu3+ ions on differentiation of hASC toward bone and cartilage forming cells. Our results showed that 10 wt% nHAp/PLLA@3 mol% Eu3+ scaffolds promotes osteogenesis and chondrogenesis of hASCs what was associated with improved synthesis and secretion of extracellular matrix proteins specific for bone and articular cartilage tissue. We also proved that obtained biomaterials have bio-imaging function and their integration with bone can be monitored using micro computed tomography (µCT).

Theranostic Applications of Nanostructured Silicate-Substituted Hydroxyapatite Codoped with Eu3+ and Bi3+ Ions-A Novel Strategy for Bone Regeneration.

In this paper, nanocrystalline silicate-substituted hydroxyapatites (nSi-HAps) codoped with Eu3+ were functionalized with Bi3+ ions. Biomaterials were synthesized using a microwave-assisted hydrothermal method and heat-treated at 700 °C. The concentration of Eu3+ ions was established at 1 mol %, and the concentration of Bi3+ was in the range of 0.5-2 mol %. The physicochemical properties of the obtained biomaterials were determined using previously established methods, including X-ray powder diffraction, scanning electron microscopy techniques, and IR spectroscopy. Particle sizes obtained in this study were in the range of 22-65 nm, which was established by the Rietveld method. The luminescence properties of the Eu3+ ion-doped silicate-substituted apatite were recorded depending on the bismuth(III) concentration. The cytocompatibility of obtained biomaterials was tested using the model of mouse pre-osteoblasts cell line, that is, MC3T3-E1. We showed that the obtained biomaterials exerted anti-apoptotic effect, reducing the number of early and late apoptotic cells and decreasing caspase activity and reactive oxygen species accumulation. The transcripts levels of genes associated with apoptosis confirmed the anti-apoptotic effect of the biomaterials. Increased metabolic activity of MC3T3-E1 in cultures with biomaterials functionalized with Bi3+ ions has been observed. Moreover, the determined profile of osteogenic markers indicates that the obtained matrices, that is, Eu3+:nSi-HAp functionalized with Bi3+ ions, exert pro-osteogenic properties. The biological features of Eu3+:nSi-HAp modified with Bi3+ ions are highly desired in terms of functional tissue restoration and further efficient osteointegration.

The Comprehensive Approach to Preparation and Investigation of the Eu3+ Doped Hydroxyapatite/poly(L-lactide) Nanocomposites: Promising Materials for Theranostics Application.

In response to the need for new materials for theranostics application, the structural and spectroscopic properties of composites designed for medical applications, received in the melt mixing process, were evaluated. A composite based on medical grade poly(L-lactide) (PLLA) and calcium hydroxyapatite (HAp) doped with Eu3+ ions was obtained by using a twin screw extruder. Pure calcium Hap, as well as the one doped with Eu3+ ions, was prepared using the precipitation method and then used as a filler. XRPD (X-ray Powder Diffraction) and IR (Infrared) spectroscopy were applied to investigate the structural properties of the obtained materials. DSC (Differential Scanning Calorimetry) was used to assess the Eu3+ ion content on phase transitions in PLLA. The tensile properties were also investigated. The excitation, emission spectra as well as decay time were measured to determine the spectroscopic properties. The simplified Judd-Ofelt (J-O) theory was applied and a detailed analysis in connection with the observed structural and spectroscopic measurements was made and described.

Study of Surface Structure Changes for Selected Ceramics Used in the CAD/CAM System on the Degree of Microbial Colonization, In Vitro Tests.

In the article has been presented an analysis of susceptibility of selected dental materials, made in the CAD/CAM technology. The morphology and structural properties of selected dental materials and their composites were determined by using XRPD (X-ray powder diffraction) techniques, as well as the IR (infrared) spectroscopy. Moreover, an adhesion as well as development of biofilm by oral microorganisms has been studied. It has been shown that a degree of the biofilm development on the tested dental materials depended on microorganism genus and species. Streptococcus mutans has demonstrated the best adhesion to the tested materials in comparison with Candida albicans and Lactobacillus rhamnosus. However, the sintered materials such as IPS e.max® and the polished IPS e.max® have showed the best "anti-adhesive properties" in relation to S. mutans and L. rhamnosus that have not formed the biofilm on the polished IPS e.max® sample. Furthermore, S. mutans have not formed the biofilm on both surfaces. On the contrary to S. mutans and L. rhamnosus, C. albicans has demonstrated the adhesive properties in relation to the above-mentioned surfaces. Moreover, in contrast to S. mutans and C. albicans, L. rhamnosus has not formed the biofilm on the polished IPS Empress material.

Nanocrystalline Hydroxyapatite Loaded with Resveratrol in Colloidal Suspension Improves Viability, Metabolic Activity and Mitochondrial Potential in Human Adipose-Derived Mesenchymal Stromal Stem Cells (hASCs).

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-α-l-galacto-ß-d-galactan (galactose hydrogel) together with resveratrol (trans-3,5,4'-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 °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.

New approach to modification of poly (l-lactic acid) with nano-hydroxyapatite improving functionality of human adipose-derived stromal cells (hASCs) through increased viability and enhanced mitochondrial activity.

The aim of this study was to determine the cytocompatibility of poly (l-lactide) (PLLA) scaffolds fabricated using co-rotating twin screw extrusion technique and functionalized with different concentrations of nano-hydroxyapatite (nHAp). The efforts were aimed on the designing bioactive scaffolds improving the viability and metabolic activity of human adipose-derived multipotent stromal cells (hASCs). The in vitro study was designed to determine the optimal nHAp concentration, based on analysis of hASCs morphology, adhesion rate, as well as metabolic and proliferative potential. Initially, the PLLA filled with three different concentrations of the nHAp were tested i.e. 5%, 10% and 15 wt%. The obtained results indicated that the 10 wt% nHAp in the PLLA (10% nHAp/PLLA) matrices improved the adhesion and proliferation of the hASCs, what was in good agreement with the results of tensile properties of the composites. Further, we performed profound studies regarding the cytotoxicity of 10% nHAp/PLLA. The analysis included the evaluation of the biomaterial influence on viability, apoptosis-related markers expression profile and mitochondrial function. The cytocompatibility of 10% nHAp/PLLA scaffolds toward the hASCs was confirmed. The hASCs propagated on 10% nHAp/PLLA were more viable then those propagated on the plain PLLA. The level of pro-apoptotic markers, i.e. caspase-3 and Bax in cultures on 10% nHAp/PLLA was significantly decreased. Obtained results correlated with higher mitochondrial membrane potential of hASCs in those cultures. The obtained composites may improve therapeutic potential of hASCs via directing their adhesion, enhancing proliferation and viability as well as increasing mitochondrial potential, thus may find potential application in tissue engineering.