Crystalline Calcium Phosphate Research Articles

Gelatin Methacryloyl (GelMA) - 45S5 Bioactive Glass (BG) Composites for Bone Tissue Engineering: 3D Extrusion Printability and Cytocompatibility Assessment Using Human Osteoblasts.

3D extrusion printing has been widely investigated for low-volume production of complex-shaped scaffolds for tissue regeneration. Gelatin methacryloyl (GelMA) is used as a baseline material for the synthesis of biomaterial inks, often with organic/inorganic fillers, to obtain a balance between good printability and biophysical properties. The present study demonstrates how 45S5 bioactive glass (BG) addition and GelMA concentrations can be tailored to develop GelMA composite scaffolds with good printability and buildability. The experimental results suggest that 45S5 BG addition consistently decreases the compression stiffness, irrespective of GelMA concentration, albeit within 20% of the baseline scaffold (without 45S5 BG). The optimal addition of 2 wt % 45S5 BG in 7.5 wt % GelMA was demonstrated to provide the best combination of printability and buildability in the 3D extrusion printing route. The degradation decreases and the swelling kinetics increases with 45S5 BG addition, irrespective of GelMA concentration. Importantly, the dissolution in simulated body fluid over 3 weeks clearly promoted the nucleation and growth of crystalline calcium phosphate particles, indicating the potential of GelMA-45S5 BG to promote biomineralization. The cytocompatibility assessment using human osteoblasts could demonstrate uncompromised cell proliferation or osteogenic marker expression over 21 days in culture for 3D printable 7.5 wt % GelMA -2 wt % 45S5 BG scaffolds when compared to 7.5 wt % GelMA. The results thus encourage further investigations of the GelMA/45S5 BG composite system for bone tissue engineering applications.

Polymer-Based Hydroxyapatite-Silver Composite Resin with Enhanced Antibacterial Activity for Dental Applications.

The primary objective of this investigation was to synthesize a resin incorporating nanoparticles of hydroxyapatite and silver (HA-NpsAg) to enhance biocompatibility and antimicrobial efficacy, thereby facilitating potential implementation within the dental industry. These enhancements aim to ensure reliable, durable, functional, and aesthetically pleasing restorations while concurrently reducing susceptibility to bacterial colonization within the oral cavity. Hydroxyapatite powders were prepared using the sol-gel method and doped with silver nanoparticles obtained by chemical reduction. The crystalline amorphous calcium phosphate powder had a particle size of 279 nm, and the silver nanoparticles had an average diameter of 26.5 nm. Resin spheres containing HA-NpsAg (RHN) were then synthesized at two concentrations (0.5% and 1%) by dissolving the initial monomer mixture in tetrahydrofuran. Subsequent antimicrobial evaluations were conducted via agar diffusion and turbidimetry, employing three strains of Gram-negative bacteria (E. coli, K. oxytoca, and P. aeruginosa) and three strains of Gram-positive bacteria (S. mutans, S. aureus, and B. subtilis). The findings revealed that P. aeruginosa exhibited maximum susceptibility to RHN powder at a concentration of 0.5%, while RHN powder at 1% concentration demonstrated maximal inhibition against S. aureus and S. mutans. Overall, our study highlights the successful synthesis of a dental resin with hydroxyapatite and silver nanoparticles, exhibiting bactericidal properties at low silver concentrations. These findings hold promise for enhancing dental materials with improved antimicrobial efficacy and clinical performance.

Dynamic Nuclear Polarization Enhanced Multiple-Quantum Spin Counting of Molecular Assemblies in Vitrified Solutions.

Crystallization pathways are essential to various industrial, geological, and biological processes. In nonclassical nucleation theory, prenucleation clusters (PNCs) form, aggregate, and crystallize to produce higher order assemblies. Microscopy and X-ray techniques have limited utility for PNC analysis due to the small size (0.5-3 nm) and time stability constraints. We present a new approach for analyzing PNC formation based on 31P nuclear magnetic resonance (NMR) spin counting of vitrified molecular assemblies. The use of glassing agents ensures that vitrification generates amorphous aqueous samples and offers conditions for performing dynamic nuclear polarization (DNP)-amplified NMR spectroscopy. We demonstrate that molecular adenosine triphosphate along with crystalline, amorphous, and clustered calcium phosphate materials formed via a nonclassical growth pathway can be differentiated from one another by the number of dipolar coupled 31P spins. We also present an innovative approach for examining spin counting data, demonstrating that a knowledge-based fitting of integer multiples of cosine wave functions, instead of the traditional Fourier transform, provides a more physically meaningful retrieval of the existing frequencies. This is the first report of multiquantum spin counting of assemblies formed in solution as captured under vitrified DNP conditions, which can be useful for future analysis of PNCs and other aqueous molecular clusters.

Equilibrium interactions of biomimetic DNA aptamers produce intrafibrillar calcium phosphate mineralization of collagen

Native and biomimetic DNA structures have been demonstrated to impact materials synthesis under a variety of conditions but have only just begun to be explored in this role compared to other biopolymers such as peptides, proteins, polysaccharides, and glycopolymers. One selected DNA aptamer has been explored in calcium phosphate and calcium carbonate mineralization, demonstrating sequence-dependent control of kinetics, morphology, and crystallinity. This aptamer is here applied to a biologically-relevant bone model system that uses collagen hydrogels. In the presence of the aptamer, intrafibrillar collagen mineralization is observed compared to negative controls and a positive control using well-studied poly-aspartic acid. The mechanism of interaction is explored through affinity measurements, kinetics of calcium uptake, and kinetics of aptamer uptake into the forming mineral. There is a marked difference observed between the selected aptamer containing a G-quadruplex secondary structure compared to a control sequence with no G-quadruplex. It is hypothesized that the equilibrium interaction of the aptamer with calcium-phosphate precursors and with the collagen itself leads to slow kinetic mineral formation and a morphology appropriate to bone. This points to new uses for DNA aptamers in biologically-relevant mineralization systems and the possibility of future biomedical applications. Statement of significanceCollagen is the protein structural component that mineralizes with calcium phosphate to form durable bone. Crystalline calcium phosphate must be infused throughout the collagen fiber structure to produce a strong material. This process is assisted by soluble proteins that interact with both calcium phosphate precursors and the collagen protein and has been proposed to follow a polymer-induce liquid precursor (PILP) model. Further understanding of this model and control of the process through synthetic, biomimetic molecules could have significant advantages in biomedical, restorative procedures. For the first time, synthetic DNA aptamers with specific secondary structures are here shown to influence and direct collagen mineralization. The mechanism of this process has been studied to demonstrate an important equilibrium between the DNA aptamer, calcium phosphate precursors, and collagen.

Polyhydroxybutyrate-based osteoinductive mineralized electrospun structures that mimic components and tissue interfaces of the osteon for bone tissue engineering.

Scaffolds for bone tissue engineering should enable regeneration of bone tissues with its native hierarchically organized ECM and multiple tissue interfaces. To achieve this, inspired by the structure and properties of bone osteon, we fabricated polyhydroxybutyrate-based mineralized electrospun fibrous scaffolds. After studying multiple polyhydroxybutyrate (PHB)-based fibers, we chose 7%PHB/1%Gelatin fibers (PG) to fabricate mineralized fibers that mimic mineralized collagen fibers in bone. The mineralized PG (mPG) surface had a rough, hydrophilic layer of low crystalline calcium phosphate which was biocompatible to BMSCs, induced their proliferation and was osteoinductive. Subsequently, by modulating the electrospinning process, we fabricated mPG-based novel higher order fibrous scaffolds that mimic the macroscale geometries of osteons of bone ECM. Inspired by the aligned collagen fibers in bone lamellae, we fabricated mPG scaffolds with aligned fibers that could direct anisotropic elongation of mBMSCs. Further, we fabricated electrospun mPG-based osteoinductive tubular constructs which can mimic cylindrical bone components like osteons or lamellae or be used as long bone analogues based on their dimensions. Finally, to regenerate tissue interfaces in bone, we introduced a novel bi-layered scaffold-based approach. An electrospun bi-layered tubular construct that had PG in the outer layer and 7%PHB/0.5%Polypyrrole fibers (PPy) in the inner layer was fabricated. The bi-layered tubular construct underwent preferential surface mineralization only on its outer layer. This outer mineralized layer supported osteogenesis while the inner PPy layer could support neural cell growth. Thus, the bi-layered tubular construct may be used to regenerate haversian canal in the osteons which hosts nerve fibers. Overall, the study introduced novel techniques to fabricate biomimetic structures that can regenerate components of bone osteon and its multiple tissue interfaces. The study lays foundation for the fabrication of a modular scaffold that can regenerate bone with its hierarchical structure and complex tissue interfaces.

Structure of biomimetic casein micelles: Critical tests of the hydrophobic colloid and multivalent-binding models using recombinant deuterated and phosphorylated β-casein

Milk contains high concentrations of amyloidogenic casein proteins and is supersaturated with respect to crystalline calcium phosphates such as apatite. Nevertheless, the mammary gland normally remains unmineralized and free of amyloid. Unlike κ-casein, β- and αS-caseins are highly effective mineral chaperones that prevent ectopic and pathological calcification of the mammary gland. Milk invariably contains a mixture of two to five different caseins that act on each other as molecular chaperones. Instead of forming amyloid fibrils, several thousand caseins and hundreds of nanoclusters of amorphous calcium phosphate combine to form fuzzy complexes called casein micelles. To understand the biological functions of the casein micelle its structure needs to be understood better than at present. The location in micelles of the highly amyloidogenic κ-casein is disputed. In traditional hydrophobic colloid models, it, alone, forms a stabilizing surface coat that also determines the average size of the micelles. In the recent multivalent-binding model, κ-casein is present throughout the micelle, in intimate contact with the other caseins. To discriminate between these models, a range of biomimetic micelles was prepared using a fixed concentration of the mineral chaperone β-casein and nanoclusters of calcium phosphate, with variable concentrations of κ-casein. A biomimetic micelle was also prepared using a highly deuterated and in vivo phosphorylated recombinant β-casein with calcium phosphate and unlabelled κ-casein. Neutron and X-ray scattering experiments revealed that κ-casein is distributed throughout the micelle, in quantitative agreement with the multivalent-binding model but contrary to the hydrophobic colloid models.

Migration of phosphorus in pig manure during pyrolysis process and slow-release mechanism of biochar in hydroponic application

Pyrolysis is an effective method for treating of livestock and poultry manure developed in recent years. It can completely decompose pathogens and antibiotics, stabilize heavy metals, and enrich phosphorus (P) in biochar. To elucidate the P migration mechanism under different pig manure pyrolysis temperatures, sequential fractionation, solution 31P nuclear magnetic resonance, X-ray photoelectron spectroscopy, X-ray diffraction, and K-edge X-ray absorption near-edge structure techniques were used to analyze the P species in pig manure biochar (PMB). The results indicated that most of the organic P in the pig manure was converted to inorganic P during pyrolysis. Moreover, the transformation to different P groups pathways was clarified. The phase transition from amorphous to crystalline calcium phosphate was promoted when the temperature was above 600 °C. The content of P extracted by hydrochloric acid, which was the long-term available P for plant uptake, increased significantly. PMB pyrolyzed at 600 °C can be used as a highly effective substitute for P source. It provides the necessary P species (e.g. water-soluble P.) and metal elements for the growth of water spinach plants, and which are slow-release comparing with the Hogland nutrient solution.

COPI Vesicle Disruption Inhibits Mineralization via mTORC1-Mediated Autophagy.

Bone mineralization is a sophisticated regulated process composed of crystalline calcium phosphate and collagen fibril. Autophagy, an evolutionarily conserved degradation system, whereby double-membrane vesicles deliver intracellular macromolecules and organelles to lysosomes for degradation, has recently been shown to play an essential role in mineralization. However, the formation of autophagosomes in mineralization remains to be determined. Here, we show that Coat Protein Complex I (COPI), responsible for Golgi-to-ER transport, plays a pivotal role in autophagosome formation in mineralization. COPI vesicles were increased after osteoinduction, and COPI vesicle disruption impaired osteogenesis. Mechanistically, COPI regulates autophagy activity via the mTOR complex 1 (mTORC1) pathway, a key regulator of autophagy. Inhibition of mTOR1 rescues the impaired osteogenesis by activating autophagy. Collectively, our study highlights the functional importance of COPI in mineralization and identifies COPI as a potential therapeutic target for treating bone-related diseases.

Titanium Dioxide Promotes the Growth and Aggregation of Calcium Phosphate and Monosodium Urate Mixed Crystals.

The increased utilization of titanium dioxide (TiO2) nanoparticles (TNPs) in various industrial and consumer products has raised concerns regarding its harmful effect due to its accumulation within the different systems of the human body. Here, we focused on the influence of TNPs on the growth and aggregation of two crucial crystalline substances, calcium phosphate (CaP) and monosodium urate (MSU), particularly its implications in gout disease. In this study, we adopted microscopic techniques and generated kinetic models to examine the interactions between TNPs, CaP and MSU, and crystallization, under controlled laboratory conditions. Our findings reveal that TNPs not only facilitate the growth of these crystals but also promote their co-aggregations. Crystal dissolution kinetics also exhibit that an increase in TNPs concentration corresponds to a reduction in the dissolution rate of CaP and MSU crystals in presence of the dissoluting agent hydroxycitrate (Hcit). These observations suggest that TNPs can stabilize CaP+MSU mixed crystals, which underscores the significance of TNPs' exposure in the pathogenesis of gout disease.

Investigation of Calcium and Magnesium Phosphate Crystals in Stones Treated with Diammonium Hydrogen Phosphate Conservation Product: Potential of Micro-Raman Spectroscopy

This study is aimed at investigating crystals (calcium and magnesium phosphates) formed due the interaction of an inorganic conservation treatment (diammonium hydrogen phosphate—DAP) with carbonatic (calcitic and dolomitic) stones through micro-Raman Spectroscopy. The addressed questions concern (i) the identification of magnesium and calcium phosphate minerals crystallized within dolomitic stone samples with a different degree of conservation state and treated with two different DAP solution molarities and (ii) the distinction of complex calcium phosphate mixtures (hydroxyapatite—HAP and octa calcium phosphate—OCP) crystallized within a calcarenite stone treated with DAP. A statistically relevant number of Raman spectra have been acquired in sample cross sections and curve fitting analysis has been performed for the in-depth interpretation of data. The outcomes indicate that Raman Spectroscopy is an effective alternative method for the identification of poorly crystalline calcium phosphates (not easily detectable with X-ray diffraction), even when scarcely present in mixture with magnesium phosphates. Evidence of the Raman analytical capability and high potential to distinguish HAP and OCP in calcitic stones are also presented and discussed.

IMPROVEMENT IN OSTEOCONDUCTIVITY OF MINISCREW IMPLANTS BY THE BIOMIMETIC CALCIUM PHOSPHATE COATINGS IN THE PRESENCE AND ABSENCE OF BOVINE SERUM ALBUMIN

Miniscrew implants (MSIs) are widely used to provide absolute anchorage for the orthodontic treatment. However, the application of MSIs is limited by the relatively high failure rate (22.86%). In this study, we wished to investigate the effects of amorphous and crystalline biomimetic calcium phosphate coating on the surfaces of MSIs with or without the incorporated BSA for the osteointegration process with an aim to facilitate the early loading of MSIs.Amorphous and crystalline coatings were prepared on titanium mini-pin implants. Characterizations of coatings were examined by Scanning electron microscopy (SEM), Confocal laser-scanning dual-channel-fluorescence microscopy (CLSM) and Fourier-transform infrared spectroscopy (FTIR). The loading and release kinetics of bovine serum albumin (BSA) were evaluated by Enzyme linked immunosorbent assay (ELISA). Activity of alkaline phosphate (ALP) was measured by using the primary osteoblasts. In vivo, a model of metaphyseal tibial implantation in rats was used (n=6 rats per group). We had 6 different groups: no coating no BSA, no coating but with surface adsorption of BSA and incorporation of BSA in the biomimetic coating in the amorphous and crystalline coatings. Time points were 3 days, 1, 2 and 4 weeks. Histological and histomorphometric analysis were performed and the bone to implant contact (BIC) of each group was compared.In vitro, the incorporation of BSA changed the crystalline coating from sharp plates into curly plates, and the crystalline coating showed slow-release profile. The incorporation of BSA in crystalline coating significantly decreased the activity of ALP in vitro. In vivo study, the earliest significant increase of BIC appeared in crystalline coating group at one week.The crystalline coating can serve as a carrier and slow release system for the bioactive agent and accelerate osteoconductivity at early stage in vivo. The presence of BSA is not favorable for the early establishment of osteointegration.

Enabling efficient phosphorus recovery from cow manure: Liberation of phosphorus through acidification and recovery of phosphorus as calcium phosphate granules

Cow manure (CM) is one of the largest secondary sources of phosphorus, while mined phosphorus rock is becoming scarce and diminishing in quality. Phosphorus recovery from CM is essential for increasing the phosphorus use efficiency and, therefore, securing future food production. Calcium phosphate (CaP) granulation can enable efficient phosphorus recovery during anaerobic digestion. However, in CM, the absence of phosphorus in the ionic form and the high presence of inorganic carbon and magnesium inhibits CaP granulation. In this study, CaP granulation was achieved for the first time during CM treatment, using an up-flow anaerobic sludge blanket (UASB) reactor preceded by an acidifying continuously stirred tank reactor (CSTR). The UASB reactor and CSTR ran for 190 days at organic loading rates of 1 and 22.5 gCOD Lreactor-1 d-1, respectively. The acidification of CM at pH 5 dissolved struvite and calcium carbonate, causing > 94% of phosphorus, calcium and magnesium to be present as ions. In the UASB reactor fed with the CSTR effluent, 83 ± 3% of phosphorus was removed and mainly present as CaP granules > 2.5 mm in diameter. The CaP granules had a phosphorus content of 8.4 ± 1.3%P w/w with amorphous and crystalline calcium phosphate being the dominant phases. The formation of large and concentrated CaP granules causes accumulation at the bottom of the reactor and allows for simple size separation leading to efficient phosphorus recovery The recovery of CaP granules improves CM valorization and can reduce the transport of manure by up to 87%. The acidification prior to digestion thus promotes a more efficient use of phosphorus from CM in agriculture and can replace phosphorus from primary phosphorus sources.

Brushite mineralised Scots pine (Pinus sylvestris L.) sapwood - revealing mineral crystallization within a wood matrix by in situ XRD.

Dicalcium phosphate dihydrate (CaHPO4·2H2O, DCPD, brushite) crystals were synthesised within Scots pine sapwood via a wet-chemistry route from aqueous solutions of Ca(CH3COO)2 and NH4H2PO4 salts. SEM/EDS analysis was used to assess the saturation of the wood cell lumina and cell wall as well as morphological features and elemental composition of the co-precipitated mineral. Brushite mineral crystallization and crystallite growth within the wood matrix was studied by in situ XRD. The chemical composition of the mineral before and after the dissolution was evaluated using FTIR spectroscopy. The overall impact of brushite on the thermal behaviour of wood was studied by TGA/DSC and TGA/DTA/MS analysis under oxidative and pyrolytic conditions. Bending and compression strength perpendicular and parallel to the fibre directions as well as bending strengths in longitudinal and transverse directions of the mineralised wood were also evaluated. Results indicate the viability of the wet-chemistry processing route for wood reinforcement with crystalline calcium phosphate (CaP)-based minerals, and imply a potential in producing hybrid bio-based materials that could be attractive in the construction sector as an environmentally friendly building material.

Molecular and biochemical approach for understanding the transition of amorphous to crystalline calcium phosphate deposits in human teeth.

Calcium phosphate (CaP) deposition during bone mineralization starts with the aggregation of Posner's clusters Ca9(PO4)6 into amorphous Ca-phosphate (ACP), which then transforms into crystalline CaP and finally maturates to hydroxyapatite (HA). Using dentin/enamel of human teeth as a model system, we show that the physiological inorganic polymer polyphosphate (polyP), a phosphate donor in mineralization, prevents the transition from amorphous to crystalline CaP at concentrations>15wt%. Stabilization of the amorphous phase of CaP by polyP is reversed by hydrolysis of the polymer by alkaline phosphatase (ALP), an enzyme that releases phosphate for mineralization. It is still present in calcified enamel and dentin, as shown here by immunostaining and enzyme activity measurements. The phase transfer into crystalline CaP can be prevented by the ALP inhibitor levamisole. Besides TEM and SEM, the modulating effects of polyP and ALP on the kinetics of the phase transition from amorphous to crystalline CaP are demonstrated and confirmed by XRD and FTIR analyses. Molecular modeling studies show that the polyP chains, due to their dimensions, are able to penetrate into the channels between the Posner molecules, preventing cluster association to ACP and impairing HA crystal formation.

Investigation of the influence of SrO addition on the bioactivity of CaO[sbnd]Na2O-P2O5-B2O3 glass system

In this work, the role of strontium oxide in the bioactive glasses calcium-sodium-borate was studied to evaluate the bioactivity of the system and the rate of the ion release in Simulated Body Fluid (SBF). Five samples were prepared, with concentration of strontium oxide (SrO) from 0 to 20 wt%, in order to understand how SrO change the glass structure and the ionic leaching process in SBF. Results of density and molar volume, XRD, FTIR, weight loss, pH and cytotoxicity assay were presented and discussed as a function of SrO. The measured pH values of SBF, presented lower values for samples with greater amounts of SrO, suggesting that the increase of SrO in the glasses can reduce the release of the ions in SBF. The fraction of tetracoordinate boron, N4, calculated by FTIR results, increases with the addition of SrO, which shows an increase in the glass connectivity and a decrease of the chemical reactivity. The FTIR spectra and X-ray diffraction results, after immersion in SBF, confirmed the materials bioactivity with the formation of carbonated hydroxyapatite or crystalline calcium phosphate. The samples did not show in vitro cytotoxicity against osteoblast cell line. Also, we highlight that 10SrO sample has induced proliferation of osteoblast-like cells in vitro to all concentrations of glass. These results show that SrO doped calcium-sodium-borate glasses are materials with high rate of bioactivity, low cytotoxicity and reduced dissolution rate of the glasses in an aqueous medium, besides induce cells proliferation and bone tissue growth and their biocompatibility with living tissues.

Calcium phosphate bioceramics: From cell behavior to chemical-physical properties

Calcium phosphate ceramics, including hydroxyapatite (HA), have been used as bone substitutes for more than 40 years. Their chemical composition, close to that of the bone mineral, confers them good biological and physical properties. However, they are not sufficient to meet all the needs in bone regenerative medicine, such as in the context of critical bone lesions. Therefore, it is essential to improve their biological performances in order to extend their application domains. In this aim, three approaches are mainly followed on the assumption that the biological response can be tuned by modifications of the chemical physical properties of the ceramic: 1) Incorporation of specific chemical species into the calcium phosphate crystalline lattice of chemical elements to stimulate bone repair. 2) Modulation of the bioceramic architecture to optimize the cellular responses at the interface. 3) Functionalization of the bioceramic surface with bioactive molecules. These approaches are supposed to act on separate parameters but, as they are implemented during different steps of the ceramic processing route, they cannot be considered as exclusive. They will ineluctably induces changes of several other physical chemical properties of the final ceramic that may also affect the biological response. Using examples of recent works from our laboratory, the present paper aims to describe how biology can be affected by the bioceramics modifications according to each one of these approaches. It shows that linking biological and chemical physical data in a rational way makes it possible to identify pertinent parameters and related processing levers to target a desired biological response and then more precisely tune the biological performance of ceramic biomaterials. This highlights the importance of integrating the biological evaluation into the heart of the processes used to manufacture optimized biomaterials.

Mineralization Profile of Annexin A6-Harbouring Proteoliposomes: Shedding Light on the Role of Annexin A6 on Matrix Vesicle-Mediated Mineralization.

The biochemical machinery involved in matrix vesicles-mediated bone mineralization involves a specific set of lipids, enzymes, and proteins. Annexins, among their many functions, have been described as responsible for the formation and stabilization of the matrix vesicles′ nucleational core. However, the specific role of each member of the annexin family, especially in the presence of type-I collagen, remains to be clarified. To address this issue, in vitro mineralization was carried out using AnxA6 (in solution or associated to the proteoliposomes) in the presence or in the absence of type-I collagen, incubated with either amorphous calcium phosphate (ACP) or a phosphatidylserine-calcium phosphate complex (PS–CPLX) as nucleators. Proteoliposomes were composed of 1,2-dipalmitoylphosphatidylcholine (DPPC), 1,2-dipalmitoylphosphatidylcholine: 1,2-dipalmitoylphosphatidylserine (DPPC:DPPS), and DPPC:Cholesterol:DPPS to mimic the outer and the inner leaflet of the matrix vesicles membrane as well as to investigate the effect of the membrane fluidity. Kinetic parameters of mineralization were calculated from time-dependent turbidity curves of free Annexin A6 (AnxA6) and AnxA6-containing proteoliposomes dispersed in synthetic cartilage lymph. The chemical composition of the minerals formed was investigated by Fourier transform infrared spectroscopy (FTIR). Free AnxA6 and AnxA6-proteoliposomes in the presence of ACP were not able to propagate mineralization; however, poorly crystalline calcium phosphates were formed in the presence of PS–CPLX, supporting the role of annexin-calcium-phosphatidylserine complex in the formation and stabilization of the matrix vesicles’ nucleational core. We found that AnxA6 lacks nucleation propagation capacity when incorporated into liposomes in the presence of PS–CPLX and type-I collagen. This suggests that AnxA6 may interact either with phospholipids, forming a nucleational core, or with type-I collagen, albeit less efficiently, to induce the nucleation process.

Modification of physicochemical properties and bioactivity of oxide coatings formed on Ti substrates via plasma electrolytic oxidation in crystalline and amorphous calcium phosphate particle suspensions

One way to improve characteristics of oxide coatings prepared by plasma electrolytic oxidation (PEO) method in suspensions lies in variation of solid particle properties. In this study, the effect of particle crystallinity on different physicochemical properties and bioactivity of coatings prepared on Ti and Ti6Al4V substrates was studied, and repeatability of resulting characteristics was assessed depending on selected substrates. PEO was performed in direct current mode at a voltage of 350 V in electrolytes containing 0.1 M KH2PO4 and 100 g.L-1 of stoichiometric crystalline hydroxyapatite or amorphous tricalcium phosphate particles with average size of ∼ 450 nm. Series of measured physical parameters included coating thickness, as well as surface wettability, roughness and topography. Surface and cross-sectional chemical composition was studied by Raman and EDX spectroscopy, while bioactivity was evaluated with simulated body fluid tests. Results showed specific influence of particle crystallinity on surface morphology and repeatable particle incorporation pattern depending on the substrate utilized. It was found that amorphous particles lead to the increased amount of incorporated calcium phosphate phases followed by improved coating bioactivity and possible explanation of this phenomenon was suggested. Variation of particle crystallinity was proposed as a new potential instrument for the adjustment of PEO coating properties.

Experimental study on the interaction of insulin with apatitic calcium phosphates analogous to bone mineral: adsorption and release

The present work investigated the interaction of human insulin with synthesized poorly crystalline apatitic calcium phosphates containing simultaneously HPO42- and CO32- ions in various amount. The adsorption kinetics is very fast, while the release kinetics is generally slow. The chemical composition of apatite has an influence on both the adsorption and release processes. The experimental results show that the percentage of insulin adsorption and release decreased with the increase of the content of carbonate. The equilibrium adsorption data are fitted into Langmuir, Freundlich, Elovich, Temkin, and Dubinin–Radushkevich isotherms. The Langmuir model is best suited with a maximum monolayer adsorption capacity of 33.20 and 25.08 mg/g at 310 K corresponding to the carbonated and octocalcium phosphate apatite respectively. Isotherms parameters have revealed that the adsorption of insulin on these apatites is a feasible, spontaneous, and exothermic process. Fourier-transforms infrared confirm the fixation of insulin on non-carbonated and carbonated apatite. The adsorption and release of insulin molecules can be well described as an ions exchange-reaction between species in the hydrated layer of apatite and other species in solution. All of these results suggested that apatitic calcium phosphates can be used as systems for insulin delivery.

Amorphous calcium phosphate, the lack of order is an abundance of possibilities

For almost three decades from its discovery, amorphous calcium phosphate (ACP) was not considered a suitable biomaterial due to its structural instability. Thanks to its unique properties in respect to crystalline calcium phosphate phases, nowadays ACP is used in promising devices for hard tissue regeneration. Here we have highlighted the features of ACP that were harnessed to create excellent biomaterials for dental remineralization, self-setting bone cements, drug delivery, and coatings of prostheses. Its current limitations as well as future perspectives of development were concisely described. Despite more research works are needed, we envisage that the future of ACP is bright.