PRECIPITATION OF HYBRID HYDROXYAPATITE / AUTOFIBRIN NANOCOMPOSITES

Синтезированы гибридные нанокомпозиты на основе гидроксиапатита и аутофибрина в форме фибринового сгустка либо цитратной плазмы путем осаждения при pH 9. «Мягкие» условия осаждения и быстрое выделение нанокомпозитов способствовали сохранению биополимерной матрицы аутофибрина. Дестабилизация дополнительной фазы аморфного фосфата кальция с образованием стехиометрического гидроксиапатита обусловлена влиянием макромолекул фибрина. Формирование кальцийдефицитного гидроксиапатита с x« 0,1 и Ca / P 1,65 происходило в среде цитратной плазмы, который после 800 °С превращался в смесь гидроксиапатит / 3 -трикальцийфосфат. Синтез композитов на основе биомиметического апатита осуществляли при добавлении 30 об.% модельного раствора Simulated Body Fluid (SBF). Влияние ионов Mg, CO~, входящих в состав SBF, способствовало стабилизации аморфного фосфата кальция и образованию карбонатзамещенного гидроксиапатита, устойчивого к термическим превращениям до 800°С. Совокупное влияние аутофибрина и ионов введенного SBF позволило управлять составом минеральной составляющей гибридных нанокомпозитов без разрушения биополимерной матрицы. Hybrid composites based on hydroxyapatite and autofibrin were synthesized by precipitation in a medium with pH = 9. Soft precipitation conditions and rapid isolation of the composite precipitates favored preservation of a biopolymer matrix of autofibrin. An effect of fibrin macromolecules contributed to destabilization of the amorphous calcium phosphate phase and formation of stoichiometric hydroxyapatite. The medium of the citrated plasma stimulated precipitation of calcium-deficient hydroxyapatite with x « 0,1 and the Ca / P ration of 1,65 which transformed into the mixture of hydroxyapatite / 3 -tricalcium phosphate at 800 °С. Biomimetic apatite composites were synthesized with an addition of 30 vol. % of a Simulated Body Fluid (SBF) model solution. The effect of Mg, CO~ ions of SBF promoted the stabilization of amorphous calcium phosphate and formation of carbonated hydroxyapatite that exhibited thermal stability up to 800 °С. The cummulative effect of autofibrin and ions of induced SBF provided controlling composition of the mineral part of hybrid nanocomposites without disruption of an autofibrin matrix.

Activated Carbon Fiber Cloth/Biomimetic Apatite: A Dual Drug Delivery System

A biomaterial that is both bioactive and capable of controlled drug release is highly attractive for bone regeneration. In previous works, we demonstrated the possibility of combining activated carbon fiber cloth (ACC) and biomimetic apatite (such as calcium-deficient hydroxyapatite (CDA)) to develop an efficient material for bone regeneration. The aim to use the adsorption properties of an activated carbon/biomimetic apatite composite to synthetize a biomaterial to be used as a controlled drug release system after implantation. The adsorption and desorption of tetracycline and aspirin were first investigated in the ACC and CDA components and then on ACC/CDA composite. The results showed that drug adsorption and release are dependent on the adsorbent material and the drug polarity/hydrophilicity, leading to two distinct modes of drug adsorption and release. Consequently, a double adsorption approach was successfully performed, leading to a multifunctional and innovative ACC-aspirin/CDA-tetracycline implantable biomaterial. In a second step, in vitro tests emphasized a better affinity of the drug (tetracycline or aspirin)-loaded ACC/CDA materials towards human primary osteoblast viability and proliferation. Then, in vivo experiments on a large cortical bone defect in rats was carried out to test biocompatibility and bone regeneration ability. Data clearly highlighted a significant acceleration of bone reconstruction in the presence of the ACC/CDA patch. The ability of the aspirin-loaded ACC/CDA material to release the drug in situ for improving bone healing was also underlined, as a proof of concept. This work highlights the possibility of bone patches with controlled (multi)drug release features being used for bone tissue repair.

Nano-imaging confirms improved apatite precipitation for high phosphate/silicate ratio bioactive glasses

Bioactive glasses convert to a biomimetic apatite when in contact with physiological solutions; however, the number and type of phases precipitating depends on glass composition and reactivity. This process is typically followed by X-ray diffraction and infrared spectroscopy. Here, we visualise surface mineralisation in a series of sodium-free bioactive glasses, using transmission electron microscopy (TEM) with energy-dispersive X-ray spectroscopy (EDXS) and X-ray nano-computed tomography (nano-CT). In the glasses, the phosphate content was increased while adding stoichiometric amounts of calcium to maintain phosphate in an orthophosphate environment in the glass. Calcium fluoride was added to keep the melting temperature low. TEM brought to light the presence of phosphate clustering and nearly crystalline calcium fluoride environments in the glasses. A combination of analytical methods, including solid-state NMR, shows how with increasing phosphate content in the glass, precipitation of calcium fluoride during immersion is superseded by fluorapatite precipitation. Nano-CT gives insight into bioactive glass particle morphology after immersion, while TEM illustrates how compositional changes in the glass affect microstructure at a sub-micron to nanometre-level.

Beyond dissolution: Xerostomia rinses affect composition and structure of biomimetic dental mineral in vitro

Xerostomia, known as dry mouth, is caused by decreased salivary flow. Treatment with lubricating oral rinses provides temporary relief of dry mouth discomfort; however, it remains unclear how their composition affects mineralized dental tissues. Therefore, the objective of this study was to analyze the effects of common components in xerostomia oral rinses on biomimetic apatite with varying carbonate contents. Carbonated apatite was synthesized and exposed to one of the following solutions for 72 hours at varying pHs: water-based, phosphorus-containing (PBS), mucin-like containing (MLC), or fluoride-containing (FC) solutions. Post-exposure results indicated that apatite mass decreased irrespective of pH and solution composition, while solution buffering was pH dependent. Raman and X-ray diffraction analysis showed that the addition of phosphorus, mucin-like molecules, and fluoride in solution decreases mineral carbonate levels and changed the lattice spacing and crystallinity of bioapatite, indicative of dissolution/recrystallization processes. The mineral recrystallized into a less-carbonated apatite in the PBS and MLC solutions, and into fluorapatite in FC. Tap water did not affect the apatite lattice structure suggesting formation of a labile carbonate surface layer on apatite. These results reveal that solution composition can have varied and complex effects on dental mineral beyond dissolution, which can have long term consequences on mineral solubility and mechanics. Therefore, clinicians should consider these factors when advising treatments for xerostomia patients.

Osteopontin regulates biomimetic calcium phosphate crystallization from disordered mineral layers covering apatite crystallites

Details of apatite formation and development in bone below the nanometer scale remain enigmatic. Regulation of mineralization was shown to be governed by the activity of non-collagenous proteins with many bone diseases stemming from improper activity of these proteins. Apatite crystal growth inhibition or enhancement is thought to involve direct interaction of these proteins with exposed faces of apatite crystals. However, experimental evidence of the molecular binding events that occur and that allow these proteins to exert their functions are lacking. Moreover, recent high-resolution measurements of apatite crystallites in bone have shown that individual crystallites are covered by a persistent layer of amorphous calcium phosphate. It is therefore unclear whether non-collagenous proteins can interact with the faces of the mineral crystallites directly and what are the consequences of the presence of a disordered mineral layer to their functionality. In this work, the regulatory effect of recombinant osteopontin on biomimetic apatite is shown to produce platelet-shaped apatite crystallites with disordered layers coating them. The protein is also shown to regulate the content and properties of the disordered mineral phase (and sublayers within it). Through solid-state NMR atomic carbon-phosphorous distance measurements, the protein is shown to be located in the disordered phases, reaching out to interact with the surfaces of the crystals only through very few sidechains. These observations suggest that non-phosphorylated osteopontin acts as regulator of the coating mineral layers and exerts its effect on apatite crystal growth processes mostly from afar with a limited number of contact points with the crystal.

Hibrid biomaterials based on hydroxyapatite and blood components

Hybrid biomaterials based on amorphous hydroxyapatite and blood components (fibrin, citrate plasma) were developed by chemical precipitation of hydroxyapatite in a biopolymer matrix (pH 11; Ca/P ratio 1.67) and by mixing 6–14 wt.% of hydroxyapatite gel (pH 7.0–7.2) with bipolymers. Chemically precipitated hydroxyapatite in biopolymer matrices is single phase or contains ticalcium phosphate impurity up to 30 %, mainly α-modification in fibrin matrix and β-modification in citrate plasma. The interaction of hydroxyapatite gel into the fibrin leads to significant amorphization of hydroxyapatite and an increase in its bioresorbability. Holding the composites with hydroxyapatite obtained by chemical precipitation in the Simulated Body Fluid model solution for 75 days leads to their partial resorption and simultaneous increase of biomimetic apatite, with its greater weight gain on composites with a fibrin. Hybrid biomaterials based on a fibrin obtained from the patient’s blood and hydroxyapatite gel showed positive result during implantation, allowing to form an adequate configuration of the defect, expanding the possibilities of ENT surgery.