New strontium-based coatings show activity against pathogenic bacteria in spine infection.

Infections of implants and prostheses represent relevant complications associated with the implantation of biomedical devices in spine surgery. Indeed, due to the length of the surgical procedures and the need to implant invasive devices, infections have high incidence, interfere with osseointegration, and are becoming increasingly difficult to threat with common therapies due to the acquisition of antibiotic resistance genes by pathogenic bacteria. The application of metal-substituted tricalcium phosphate coatings onto the biomedical devices is a promising strategy to simultaneously prevent bacterial infections and promote osseointegration/osseoinduction. Strontium-substituted tricalcium phosphate (Sr-TCP) is known to be an encouraging formulation with osseoinductive properties, but its antimicrobial potential is still unexplored. To this end, novel Sr-TCP coatings were manufactured by Ionized Jet Deposition technology and characterized for their physiochemical and morphological properties, cytotoxicity, and bioactivity against Escherichia coli ATCC 8739 and Staphylococcus aureus ATCC 6538P human pathogenic strains. The coatings are nanostructured, as they are composed by aggregates with diameters from 90 nm up to 1 µm, and their morphology depends significantly on the deposition time. The Sr-TCP coatings did not exhibit any cytotoxic effects on human cell lines and provided an inhibitory effect on the planktonic growth of E. coli and S. aureus strains after 8 h of incubation. Furthermore, bacterial adhesion (after 4 h of exposure) and biofilm formation (after 24 h of cell growth) were significantly reduced when the strains were cultured on Sr-TCP compared to tricalcium phosphate only coatings. On Sr-TCP coatings, E. coli and S. aureus cells lost their organization in a biofilm-like structure and showed morphological alterations due to the toxic effect of the metal. These results demonstrate the stability and anti-adhesion/antibiofilm properties of IJD-manufactured Sr-TCP coatings, which represent potential candidates for future applications to prevent prostheses infections and to promote osteointegration/osteoinduction.

Strontium and Copper Co-Doped Multifunctional Calcium Phosphates: Biomimetic and Antibacterial Materials for Bone Implants.

ß-tricalcium phosphate (ß-TCP) is a promising material in regenerative traumatology for the creation of bone implants. Previously, it was established that doping the structure with certain cations can reduce the growth of bacterial activity. Recently, much attention has been paid to co-doped ß-TCP, that is explained by their ability, on the one hand, to reduce cytotoxicity for cells of the human organism, on the other hand, to achieve a successful antibacterial effect. Sr, Cu-co-doped solid solutions of the composition Ca9.5-xSrxCu(PO4)7 was obtained by the method of solid-phase reactions. The Rietveld method of structural refinement revealed the presence of Sr2+ ions in four crystal sites: M1, M2, M3, and M4. The M5 site is completely occupied by Cu2+. Isomorphic substitution of Ca2+ → (Sr2+and Cu2+) expands the concentration limits of the existence of the solid solution with the ß-TCP structure. No additional phases were formed up to x = 4.5 in Ca9.5-xSrxCu(PO4)7. Biocompatibility tests were performed on cell lines of human bone marrow mesenchymal stromal cells (hMSC), human fibroblasts (MRC-5) and osteoblasts (U-2OS). It was demonstrated that cytotoxicity exhibited a concentration dependence, along with an increase in osteogenesis and cell proliferation. Ca9.5-xSrxCu(PO4)7 powders showed significant inhibitory activity against pathogenic strains Escherichia coli and Staphylococcus aureus. Piezoelectric properties of Ca9.5-xSrxCu(PO4)7 were investigated. Possible ways to achieve high piezoelectric response are discussed. The combination of bioactive properties of Ca9.5-xSrxCu(PO4)7 renders them multifunctional materials suitable for bone substitutes.

Exploring Borate-Modified Calcium Phosphate Ceramics: Antimicrobial Potential and Cytocompatibility Assessment.

Addressing periprosthetic infections, which present significant healing challenges that often require revision surgeries, necessitates the development of novel antibacterial materials and implants. Current research focuses on creating materials that hinder bacterial adhesion, colonization, and proliferation in surrounding tissues. Boron (B)-containing compounds are known for their antibacterial properties and potential in bone metabolism for regenerative medicine. In this study, we synthesized B-containing tricalcium phosphate (0.3B-TCP) with 1.1 wt.% B content via precipitation from aqueous solutions and sintering at 1100 °C. X-ray diffraction confirmed the ceramic's primary crystalline phase as ß-TCP, with B evenly distributed according to energy-dispersive spectroscopy data. Electron paramagnetic resonance (EPR) data verified stable paramagnetic borate anions, indicating successful BO33- substitution for phosphate groups. The microstructural properties of 0.3B-TCP ceramic were assessed before and after soaking in a saline solution. Its bending strength was approximately 30 MPa, and its porosity was about 33%. 0.3B-TCP ceramic demonstrated significant antimicrobial efficacy against various bacterial strains and a fungus. Cytotoxicity evaluation using equine adipose tissue-derived mesenchymal stem cells and osteogenic differentiation assessment were conducted. The combination of antibacterial efficacy and good cytocompatibility suggests 0.3B-TCP ceramic as a promising bone substitute material.

Polyvinylpyrrolidone-Alginate Film Barriers for Abdominal Surgery: Anti-Adhesion Effect in Murine Model.

Surgical operations on the peritoneum are often associated with the formation of adhesions, which can interfere with the normal functioning of the internal organs. The effectiveness of existing barrier materials is relatively low. In this work, the effectiveness of soluble alginate-polyvinylpyrrolidone (PVP-Alg) and non-soluble Ca ion cross-linked (PVP-Alg-Ca) films in preventing these adhesions was evaluated. Experiments in vivo were performed on mice via mechanical injury to the adjacent peritoneum wall and the caecum, followed by the application of PVP-Alg or PVP-Alg-Ca films to the injured area. After 7 days, samples from the peritoneal wall and caecum were analyzed using histology and quantitative polymerase chain reaction (qPCR). It was shown that the expression of genes responsible for adhesion formation in the caecum in the PVP-Alg group was comparable to that in the control group, while in the PVP-Alg-Ca group, it increased by 5-10 times. These results were consistent with the histology: in the PVP-Alg group, the adhesions did not form, while in the PVP-Alg-Ca group, the adhesions corresponded to five points on the adhesion scale. Therefore, the formation of intraperitoneal adhesions can be effectively prevented by non-crosslinked, biodegradable PVP-Alg films, whereas cross-linked, not biodegradable PVP-Alg-Ca films cause inflammation and adhesion formation.

Polyvinylpyrrolidone-Alginate-Carbonate Hydroxyapatite Porous Composites for Dental Applications.

An alternative approach for the currently used replacement therapy in dentistry is to apply materials that restore tooth tissue. Among them, composites, based on biopolymers with calcium phosphates, and cells can be applied. In the present work, a composite based on polyvinylpyrrolidone (PVP) and alginate (Alg) with carbonate hydroxyapatite (CHA) was prepared and characterized. The composite was investigated by X-ray diffraction, infrared spectroscopy, electron paramagnetic resonance (EPR) and scanning electron microscopy methods, and the microstructure, porosity, and swelling properties of the material were described. In vitro studies included the MTT test using mouse fibroblasts, and adhesion and survivability tests with human dental pulp stem cells (DPSC). The mineral component of the composite corresponded to CHA with an admixture of amorphous calcium phosphate. The presence of a bond between the polymer matrix and CHA particles was shown by EPR. The structure of the material was represented by micro- (30-190 µm) and nano-pores (average 8.71 ± 4.15 nm). The swelling measurements attested that CHA addition increased the polymer matrix hydrophilicity by 200%. In vitro studies demonstrated the biocompatibility of PVP-Alg-CHA (95 ± 5% cell viability), and DPSC located inside the pores. It was concluded that the PVP-Alg-CHA porous composite is promising for dentistry applications.

Antibacterial Composite Material Based on Polyhydroxybutyrate and Zn-Doped Brushite Cement.

A composite material based on electrospinning printed polyhydroxybutyrate fibers impregnated with brushite cement containing Zn substitution was developed for bone implant applications. Powder X-ray Diffraction (PXRD), Fourier Transform Infrared Spectroscopy and Scanning Electron Microscopy were applied for materials characterization. Soaking the composite in Ringer's solution led to the transformation of brushite into apatite phase, accompanied by the morphology changes of the material. The bending strength of the composite material was measured to be 3.1 ± 0.5 MPa. NCTC mouse fibroblast cells were used to demonstrate by means of the MTT test that the developed material was not cytotoxic. The behavior of the human dental pulp stem cells on the surface of the composite material investigated by the direct contact method was similar to the control. It was found that the developed Zn containing composite material possessed antibacterial properties, as testified by microbiology investigations against bacteria strains of Escherichia coli and Staphylococcus aureus. Thus, the developed composite material is promising for the treatment of damaged tissues with bacterial infection complications.

1H NMR and EPR Spectroscopies Investigation of Alginate Cross-Linking by Divalent Ions.

Alginate is a natural polymer widely applied in materials science, medicine, and biotechnology. Its ability to bind metal ions in order to form insoluble gels has been comprehensively used to create capsules for cell technology, drug delivery, biomedical materials, etc. To modify and predict the properties of cross-linked alginate, knowledge about the mechanism of alginate binding with metal ions and the properties of its gels is necessary. This article presents the results obtained by proton Nuclear Magnetic Resonance Spectroscopy for alginate containing calcium and strontium (alkaline earth metal diamagnetic) ions and by Electron Paramagnetic Resonance Spectroscopy for alginate with copper (Cu) and manganese (Mn) (transition metal paramagnetic) ions. It was found that in the case of calcium (Ca) and Mn ions, their concentration does not affect their distribution in the alginate structure and the cross-linking density. In the case of strontium (Sr) and Cu ions, their number affects the number of binding sites and, accordingly, the cross-linking density. Thus, the cross-linking of alginate depends mainly on the characteristics of specific cations, while the nature of the bond (ionic or coordination type) is less important.

Antimicrobial and Cell-Friendly Properties of Cobalt and Nickel-Doped Tricalcium Phosphate Ceramics.

ß-Tricalcium phosphate (ß-TCP) is widely used as bone implant material. It has been observed that doping the ß-TCP structure with certain cations can help in combating bacteria and pathogenic microorganisms. Previous literature investigations have focused on tricalcium phosphate structures with silver, copper, zinc, and iron cations. However, there are limited studies available on the biological properties of ß-TCP containing nickel and cobalt ions. In this work, Ca10.5-xNix(PO4)7 and Ca10.5-xCox(PO4)7 solid solutions with the ß-Ca3(PO4)2 structure were synthesized by a high-temperature solid-state reaction. Structural studies revealed the ß-TCP structure becomes saturated at 9.5 mol/% for Co2+ or Ni2+ ions. Beyond this saturation point, Ni2+ and Co2+ ions form impurity phases after complete occupying of the octahedral M5 site. The incorporation of these ions into the ß-TCP crystal structure delays the phase transition to the α-TCP phase and stabilizes the structure as the temperature increases. Biocompatibility tests conducted on adipose tissue-derived mesenchymal stem cells (aMSC) using the (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) (MTT) assay showed that all prepared samples did not exhibit cytotoxic effects. Furthermore, there was no inhibition of cell differentiation into the osteogenic lineage. Antibacterial properties were studied on the C. albicans fungus and on E. coli, E. faecalis, S. aureus, and P. aeruginosa bacteria strains. The Ni- and Co-doped ß-TCP series exhibited varying degrees of bacterial growth inhibition depending on the doping ion concentration and the specific bacteria strain or fungus. The combination of antibacterial activity and cell-friendly properties makes these phosphates promising candidates for anti-infection bone substitute materials.

Strontium Substituted ß-Tricalcium Phosphate Ceramics: Physiochemical Properties and Cytocompatibility.

Sr2+-substituted ß-tricalcium phosphate (ß-TCP) powders were synthesized using the mechano-chemical activation method with subsequent pressing and sintering to obtain ceramics. The concentration of Sr2+ in the samples was 0 (non-substituted TCP, as a reference), 3.33 (0.1SrTCP), and 16.67 (0.5SrTCP) mol.% with the expected Ca3(PO4)2, Ca2.9Sr0.1(PO4)2, and Ca2.5Sr0.5(PO4)2 formulas, respectively. The chemical compositions were confirmed by the energy-dispersive X-ray spectrometry (EDX) and the inductively coupled plasma optical emission spectroscopy (ICP-OES) methods. The study of the phase composition of the synthesized powders and ceramics by the powder X-ray diffraction (PXRD) method revealed that ß-TCP is the main phase in all compounds except 0.1SrTCP, in which the apatite (Ap)-type phase was predominant. TCP and 0.5SrTCP ceramics were soaked in the standard saline solution for 21 days, and the phase analysis revealed the partial dissolution of the initial ß-TCP phase with the formation of the Ap-type phase and changes in the microstructure of the ceramics. The Sr2+ ion release from the ceramic was measured by the ICP-OES. The human osteosarcoma MG-63 cell line was used for viability, adhesion, spreading, and cytocompatibility studies. The results show that the introduction of Sr2+ ions into the ß-TCP improved cell adhesion, proliferation, and cytocompatibility of the prepared samples. The obtained results provide a base for the application of the Sr2+-substituted ceramics in model experiments in vivo.

Influence of Synthesis Conditions on Gadolinium-Substituted Tricalcium Phosphate Ceramics and Its Physicochemical, Biological, and Antibacterial Properties.

Gadolinium-containing calcium phosphates are promising contrast agents for various bioimaging modalities. Gadolinium-substituted tricalcium phosphate (TCP) powders with 0.51 wt% of gadolinium (0.01Gd-TCP) and 5.06 wt% of (0.1Gd-TCP) were synthesized by two methods: precipitation from aqueous solutions of salts (1) (Gd-TCP-pc) and mechano-chemical activation (2) (Gd-TCP-ma). The phase composition of the product depends on the synthesis method. The product of synthesis (1) was composed of ß-TCP (main phase, 96%), apatite/chlorapatite (2%), and calcium pyrophosphate (2%), after heat treatment at 900 °C. The product of synthesis (2) was represented by ß-TCP (main phase, 73%), apatite/chlorapatite (20%), and calcium pyrophosphate (7%), after heat treatment at 900 °C. The substitution of Ca2+ ions by Gd3+ in both ß-TCP (main phase) and apatite (admixture) phases was proved by the electron paramagnetic resonance technique. The thermal stability and specific surface area of the Gd-TCP powders synthesized by two methods were significantly different. The method of synthesis also influenced the size and morphology of the prepared Gd-TCP powders. In the case of synthesis route (1), powders with particle sizes of tens of nanometers were obtained, while in the case of synthesis (2), the particle size was hundreds of nanometers, as revealed by transmission electron microscopy. The Gd-TCP ceramics microstructure investigated by scanning electron microscopy was different depending on the synthesis route. In the case of (1), ceramics with grains of 1-50 µm, pore sizes of 1-10 µm, and a bending strength of about 30 MPa were obtained; in the case of (2), the ceramics grain size was 0.4-1.4 µm, the pore size was 2 µm, and a bending strength of about 39 MPa was prepared. The antimicrobial activity of powders was tested for four bacteria (S. aureus, E. coli, S. typhimurium, and E. faecalis) and one fungus (C. albicans), and there was roughly 30% of inhibition of the micro-organism's growth. The metabolic activity of the NCTC L929 cell and viability of the human dental pulp stem cell study demonstrated the absence of toxic effects for all the prepared ceramic materials doped with Gd ions, with no difference for the synthesis route.

Composite Polyvinylpyrrolidone-Sodium Alginate-Hydroxyapatite Hydrogel Films for Bone Repair and Wound Dressings Applications.

Today, the synthesis of biocompatible and bioresorbable composite materials such as "polymer matrix-mineral constituent," which stimulate the natural growth of living tissues and the restoration of damaged parts of the body, is one of the challenging problems in regenerative medicine. In this study, composite films of bioresorbable polymers of polyvinylpyrrolidone (PVP) and sodium alginate (SA) with hydroxyapatite (HA) were obtained. HA was introduced by two different methods. In one of them, it was synthesized in situ in a solution of polymer mixture, and in another one, it was added ex situ. Phase composition, microstructure, swelling properties and biocompatibility of films were investigated. The crosslinked composite PVP-SA-HA films exhibit hydrogel swelling characteristics, increasing three times in mass after immersion in a saline solution. It was found that composite PVP-SA-HA hydrogel films containing HA synthesized in situ exhibited acute cytotoxicity, associated with the presence of HA synthesis reaction byproducts-ammonia and ammonium nitrate. On the other hand, the films with HA added ex situ promoted the viability of dental pulp stem cells compared to the films containing only a polymer PVP-SA blend. The developed composite hydrogel films are recommended for such applications, such as membranes in osteoplastic surgery and wound dressing.

Antibacterial and cell-friendly copper-substituted tricalcium phosphate ceramics for biomedical implant applications.

The development of new materials with antibacterial properties and the scope to decrease or eliminate the excessive antibiotic use is an urgent priority due to the growing antibiotic resistance-related mortalities. New bone substitute materials with intrinsic antibacterial characteristics are highly requested for various clinical applications. In this study, the choice of copper ions as substitutes for calcium in tricalcium phosphate (TCP) has been justified by their pronounced broad-spectrum antibacterial properties. Copper-substituted TCP (Cu-TCP) ceramics with the copper content of 1.4 and 0.1 wt% were synthesized by mechano-chemical activation. X-ray diffraction (XRD) analyses established that both pure and copper-containing compounds adopted the structure of whitlockite (ß-TCP). XRD and electron paramagnetic resonance (EPR) spectroscopy revealed the partial isovalent substitution of calcium ions with copper ions in the ß-TCP lattice. With the use of infrared and EPR spectroscopies, it was detected that carbonate ions got incorporated into the ß-TCP structure during the synthesis procedure. By releasing the tension in the M(5)O6 octahedron consequential to the lower CaO bond length than the corresponding sum of ionic radii, the substitution of calcium with smaller copper ions stabilizes the structure of ß-TCP. As concluded form the thermal analyses, the introduction of Cu prevented the polymorphic transformation of ß- to α-TCP. At the same time, the introduction of Cu to the ß-TCP structure enhanced the crystal growth and porosity of the ceramics, which had a positive effect on the cytocompatibility of the material. The MTT colorimetric assay showed that the metabolic activity of the mouse fibroblast NCTC L929 cell line during 24 h of incubation with 3-day extracts from Cu-TCP (1.4 wt%) and ß-TCP pellets in the cell culture medium was similar to the negative control, indicating the absence of any inhibitory effects on cells. The seeding and the growth of human dental pulp stem cells on the surface of Cu-TCP (1.4 wt%) and ß-TCP ceramics also showed the absence of any signs of cytotoxicity. Finally, microbiological assays demonstrated the antibacterial activity of Cu-TCP ceramics against Escherichia coli and Salmonella enteritidis, whereas ß-TCP did not exhibit such an activity. Overall, the addition of Cu ions to ß-TCP improves its antibacterial properties without diminishing the biocompatibility of the material, thus making it more attractive than pure ß-TCP for clinical applications such as synthetic bone grafts and orthopaedic implant coatings.

Improved cytocompatibility and antibacterial properties of zinc-substituted brushite bone cement based on ß-tricalcium phosphate.

For bone replacement materials, osteoconductive, osteoinductive, and osteogenic properties are desired. The bacterial resistance and the need for new antibacterial strategies stand among the most challenging tasks of the modern medicine. In this work, brushite cements based on powders of Zinc (Zn) (1.4 wt%) substituted tricalcium phosphate (ß-TCP) and non-substituted ß-TCP were prepared and investigated. Their initial and final phase composition, time of setting, morphology, pH evolution, and compressive strength are reported. After soaking for 60 days in physiological solution, the cements transformed into a mixture of brushite and hydroxyapatite. Antibacterial activity of the cements against Enterococcus faecium, Escherichia coli, and Pseudomonas aeruginosa bacteria strains was attested. The absence of cytotoxicity of cements was proved for murine fibroblast NCTC L929 cells. Moreover, the cell viability on the ß-TCP cement containing Zn2+ ions was 10% higher compared to the ß-TCP cement without zinc. The developed cements are perspective for applications in orthopedics and traumatology.

Ionized jet deposition of antimicrobial and stem cell friendly silver-substituted tricalcium phosphate nanocoatings on titanium alloy.

Orthopedic infections pose severe societal and economic burden and interfere with the capability of the implanted devices to integrate in the host bone, thus significantly increasing implants failure rate. To address infection and promote integration, here nanostructured antibacterial and bioactive thin films are proposed, obtained, for the first time, by Ionized Jet Deposition (IJD) of silver-substituted tricalcium phosphate (Ag-TCP) targets on titanium. Coatings morphology, composition and mechanical properties are characterized and proof-of-concept of biocompatibility is shown. Antimicrobial efficacy is investigated against four Gram positive and Gram negative bacterial strains and against C. albicans fungus, by investigating the modifications in planktonic bacterial growth in the absence and presence of silver. Then, for all bacterial strains, the capability of the film to inhibit bacterial adhesion is also tested. Results indicate that IJD permits a fine control over films composition and morphology and deposition of films with suitable mechanical properties. Biological studies show a good efficacy against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Enterococcus faecalis and against fungus Candida albicans, with evidences of efficacy against planktonic growth and significant reduction of bacterial cell adhesion. No cytotoxic effects are evidenced for equine adipose tissue derived mesenchymal stem cells (ADMSCs), as no reductions are caused to cells viability and no interference is assessed in cells differentiation towards osteogenic lineage, in the presence of silver. Instead, thanks to nanostructuration and biomimetic composition, tricalcium phosphate (TCP) coatings favor cells viability, also when silver-substituted. These findings show that silver-substituted nanostructured coatings are promising for orthopedic implant applications.

In Vitro Properties of Manganese-Substituted Tricalcium Phosphate Coatings for Titanium Biomedical Implants Deposited by Arc Plasma.

Bioactive manganese (Mn)-doped ceramic coatings for intraosseous titanium (Ti) implants are developed. Arc plasma deposition procedure is used for coatings preparation. X-ray Diffraction, Scanning Electron Microscopy-Energy Dispersive X-ray Spectroscopy, and Electron Paramagnetic Resonance (EPR) methods are applied for coatings characterization. The coatings are homogeneous, composed of the main phase α-tricalcium phosphate (α-TCP) (about 67%) and the minor phase hydroxyapatite (about 33%), and the Mn content is 2.3 wt%. EPR spectroscopy demonstrates that the Mn ions are incorporated in the TCP structure and are present in the coating in Mn2+ and Mn3+ oxidation states, being aggregated in clusters. The wetting contact angle of the deposited coatings is suitable for cells' adhesion and proliferation. In vitro soaking in physiological solution for 90 days leads to a drastic change in phase composition; the transformation into calcium carbonate and octacalcium phosphate takes place, and no more Mn is present. The absence of antibacterial activity against Escherichia coli, Enterococcus faecalis, and Pseudomonas aeruginosa bacteria strains is observed. A study of the metabolic activity of mouse fibroblasts of the NCTC L929 cell line on the coatings using the MTT (dye compound 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) test demonstrates that there is no toxic effect on the cell culture. Moreover, the coating material supports the adhesion and proliferation of the cells. A good adhesion, spreading, and proliferative activity of the human tooth postnatal dental pulp stem cells (DPSC) is demonstrated. The developed coatings are promising for implant application in orthopedics and dentistry.

Tricalcium phosphate cement supplemented with boron nitride nanotubes with enhanced biological properties.

A self-setting bone cement containing ß-tricalcium phosphate (TCP) supplemented with boron nitride nanotubes (BNNTs, 1 wt%) was synthesized and analyzed in situ for its kinetics of hardening and selected physicochemical and biological properties. Moderately delayed due to the presence of BNNTs, the hardening reaction involved the transformation of the TCP precursor to the dicalcium phosphate (DCPD) product. In spite of the short-lived chemical transformations in the cement upon its hardening, the structural changes in it were extended. As a result, the compressive strength increased from day 1 to day 7 of the hardening reaction and the presence of BNNTs further increased it by ~25%. Fitting of the time-resolved energy-dispersive diffractometric data to the Johnson-Mehl-Avrami-Kolmogorov crystallization kinetics model conformed to the one-dimensional nucleation at a variable rate during the growth of elongated DCPD crystals from round TCP grains. For the first seven days of growth of human mesenchymal stem cells (hMSCs) on the cement, no difference in their proliferation was observed compared to the control. However, between the 7th and the 21st day, the cell proliferation decreased compared to the control because of the ongoing stem cell differentiation toward the osteoblast phenotype. This differentiation was accompanied by the higher expression of alkaline phosphatase, an early marker of hMSC differentiation into a pre-osteoblast phenotype. The TCP cement supplemented with BNNTs was able to thwart the production of reactive oxygen species (ROS) in hMSCs treated with H2O2/Fe2+ and bring the ROS levels down to the concentrations detected in the control cells, indicating the good capability of the material to protect the cells against the ROS-associated damage. Simultaneously, the cement increased the expression of mediators of inflammation in a co-culture of osteoblasts and macrophages, thus attesting to the direct reciprocity between the degrees of inflammation and stimulated new bone production.

Comparative Study of the Structure, Properties, and Corrosion Behavior of Sr-Containing Biocoatings on Mg0.8Ca.

A comparative analysis of the structure, properties and the corrosion behavior of the micro-arc coatings based on Sr-substituted hydroxyapatite (Sr-HA) and Sr-substituted tricalcium phosphate (Sr-TCP) deposited on Mg0.8Ca alloy substrates was performed. The current density during the formation of the Sr-HA coatings was higher than that for the Sr-TCP coatings. As a result, the Sr-HA coatings were thicker and had a greater surface roughness Ra than the Sr-TCP coatings. In addition, pore sizes of the Sr-HA were almost two times larger. The ratio (Ca + Sr + Mg)/P were equal 1.64 and 1.47 for Sr-HA and Sr-TCP coatings, respectively. Thus, it can be assumed that the composition of Sr-HA and Sr-TCP coatings was predominantly presented by (Sr,Mg)-substituted hydroxyapatite and (Sr,Mg)-substituted tricalcium phosphate. However, the average content of Sr was approximately the same for both types of the coatings and was equal to 1.8 at.%. The Sr-HA coatings were less soluble and had higher corrosion resistance than the Sr-TCP coatings. Cytotoxic tests in vitro demonstrated a higher cell viability after cultivation with extracts of the Sr-HA coatings.

Modification of PMMA Cements for Cranioplasty with Bioactive Glass and Copper Doped Tricalcium Phosphate Particles.

Cranioplasty represents the surgical repair of bone defects or deformities in the cranium arising from traumatic skull bone fracture, cranial bone deformities, bone cancer, and infections. The actual gold standard in surgery procedures for cranioplasty involves the use of biocompatible materials, and repair or regeneration of large cranial defects is particularly challenging from both a functional and aesthetic point of view. PMMA-based bone cement are the most widely biomaterials adopted in the field, with at least four different surgical approaches. Modifications for improving biological and mechanical functions of PMMA-based bone cement have been suggested. To this aim, the inclusion of antibiotics to prevent infection has been shown to provide a reduction of mechanical properties in bending. Therefore, the development of novel antibacterial active agents to overcome issues related to mechanical properties and bacterial resistance to antibiotics is still encouraged. In this context, mechanical, biological, and antibacterial feature against P. aeruginosa and S. aureus bacterial strains of surgical PMMA cement modified with BG and recently developed Cu-TCP bioactive particles have been highlighted.

Sic Parvis Magna: Manganese-Substituted Tricalcium Phosphate and Its Biophysical Properties.

Succeeding in the substitution of pharmaceutical compounds with ions deliverable with the use of resorbable biomaterials could have far-reaching benefits for medicine and economy. Calcium phosphates are known as excellent accommodators of foreign ions. Manganese, the fifth most abundant metal on Earth was studied here as an ionic dopant in ß-tricalcium phosphate (ß-TCP) ceramics. ß-TCP containing different amounts of Mn2+ ions per MnxCa3-x(PO4)2 formula (x = 0, 0.001, 0.01, and 0.1) was investigated for a range of physicochemical and biological properties. The results suggested the role of Mn2+ as a structure booster, not breaker. Mn2+ ions increased the size of coherent X-ray scattering regions averaged across all crystallographic directions and also lowered the temperature of transformation of the hydroxyapatite precursor to ß-TCP. The particle size increased fivefold, from 20 to 100 nm, in the 650-750 °C region, indicating that the reaction of formation of ß-TCP was accompanied by a considerable degree of grain growth. The splitting of the antisymmetric stretching mode of the phosphate tetrahedron occurred proportionally to the Mn2+ content in the material, while electron paramagnetic resonance spectra suggested that Mn2+ might substitute for three out of five possible calcium ion positions in the unit cell of ß-TCP. The biological effects of Mn-free ß-TCP and Mn-doped ß-TCP were selective: moderately proliferative to mammalian cells, moderately inhibitory to bacteria, and insignificant to fungi. Unlike pure ß-TCP, ß-TCP doped with the highest concentration of Mn2+ ions significantly inhibited the growth of all bacterial species tested: Staphylococcus aureus, Salmonella typhi, Escherichia coli, Pseudomonas aeruginosa, and Enterococcus faecalis. The overall effect against the Gram-positive bacteria was more intense than against the Gram-negative microorganisms. Meanwhile, ß-TCP alone had an augmentative effect of the viability of adipose-derived mesenchymal stem cells (ADMSCs) and the addition of Mn2+ tended to reduce the extent of this augmentative effect, but without imparting any toxicity. For all Mn-doped ß-TCP concentrations except the highest, the cell viability after 72 h incubation was significantly higher than that of the negative control. Assays evaluating the effect of Mn2+-containing ß-TCP formulations on the differentiation of ADMSCs into three different lineages-osteogenic, adipogenic, and chondrogenic-demonstrated no inhibitory or adverse effects compared to pure ß-TCP and powder-free positive controls. Still, ß-TCP delivering the lowest amount of Mn2+ seemed most effective in sustaining the differentiation process toward all three phenotypes, indicating that the dose of Mn2+ in ß-TCP need not be excessive to be effective.

Gold is for the mistress, silver for the maid: Enhanced mechanical properties, osteoinduction and antibacterial activity due to iron doping of tricalcium phosphate bone cements.

Self-hardening calcium phosphate cements present ideal bone tissue substitutes from the standpoints of bioactivity and biocompatibility, yet they suffer from (a) weak mechanical properties, (b) negligible osteoinduction without the use of exogenous growth factors, and (c) a lack of intrinsic antibacterial activity. Here we attempt to improve on these deficiencies by studying the properties of self-setting Fe-doped bone-integrative cements containing two different concentrations of the dopant: 0.49 and 1.09 wt% Fe. The hardening process, which involved the transformation of Fe-doped ß-tricalcium phosphate (Fe-TCP) to nanocrystalline brushite, was investigated in situ by continuously monitoring the cements using the Energy Dispersive X-Ray Diffraction technique. The setting time was 20 min and the hardening time 2 h, but it took 50 h for the cement to completely stabilize compositionally and mechanically. Still, compared to other similar systems, the phase transformation during hardening was relatively fast and it also followed a relatively simple reaction path, virtually free of complex intermediates and noisy background. Mössbauer spectrometry demonstrated that 57Fe atoms in Fe-TCP were located in two non-equivalent crystallographic sites and distributed over positions with a strong crystal distortion. The pronounced presence of ultrafine crystals in the final, brushite phase contributed to the reduction of the porosity and thereby to the enhancement of the mechanical properties. The compressive strength of the hardened TCP cements increased by more than twofold when Fe was added as a dopant, i.e., from 11.5 ±â€¯0.5 to 24.5 ±â€¯2.0 MPa. The amount of iron released from the cements in physiological media steadied after 10 days and was by an order of magnitude lower than the clinical threshold that triggers the toxic response. The cements exhibited osteoinductive activity, as observed from the elevated levels of expression of genes encoding for osteocalcin and Runx2 in both undifferentiated and differentiated MC3T3-E1 cells challenged with the cements. The osteoinductive effect was inversely proportional to the content of Fe ions in the cements, indicating that an excessive amount of iron can have a detrimental effect on the induction of bone growth by osteoblasts in contact with the cement. In contrast, the antibacterial activity of the cement in the agar assay increased against all four bacterial species analysed (E. coli, S. enteritidis, P. aeruginosa, S. aureus) in direct proportion with the concentration of Fe ions in it, indicating their key effect on the promotion of the antibacterial effect in this material. This effect was less pronounced in broth assays. Experiments involving co-incubation of cements with cells in an alternate magnetic radiofrequency field for 30 min demonstrated a good potential for the use of these magnetic cements in hyperthermia cancer therapies. Specifically, the population of human glioblastoma cells decreased six-fold at the 24 h time point following the end of the magnetic field treatment, while the population of the bone cancer cells dropped approximately twofold. The analysis of the MC3T3-E1 cell/cement interaction reiterated the effects of iron in the cement on the bone growth marker expression by showing signs of adverse effects on the cell morphology and proliferation only for the cement containing the higher concentration of Fe ions (1.09 wt%). Biological testing concluded that the effects of iron are beneficial from the perspective of a magnetic hyperthermia therapy and antibacterial prophylaxis, but its concentration in the material must be carefully optimized to avoid the adverse effects induced above a certain level of iron concentrations.