Enhanced Release of Calcium Ions from Hydroxyapatite Nanoparticles with an Increase in Their Specific Surface Area.

Synthetic calcium phosphates, e.g., hydroxyapatite (HAP) and tricalcium phosphate (TCP), are the most commonly used bone-graft materials due to their high chemical similarity to the natural hydroxyapatite-the inorganic component of bones. Calcium in the form of a free ion or bound complexes plays a key role in many biological functions, including bone regeneration. This paper explores the possibility of increasing the Ca2+-ion release from HAP nanoparticles (NPs) by reducing their size. Hydroxyapatite nanoparticles were obtained through microwave hydrothermal synthesis. Particles with a specific surface area ranging from 51 m2/g to 240 m2/g and with sizes of 39, 29, 19, 11, 10, and 9 nm were used in the experiment. The structure of the nanomaterial was also studied by means of helium pycnometry, X-ray diffraction (XRD), and transmission-electron microscopy (TEM). The calcium-ion release into phosphate-buffered saline (PBS) was studied. The highest release of Ca2+ ions, i.e., 18 mg/L, was observed in HAP with a specific surface area 240 m2/g and an average nanoparticle size of 9 nm. A significant increase in Ca2+-ion release was also observed with specific surface areas of 183 m2/g and above, and with nanoparticle sizes of 11 nm and below. No substantial size dependence was observed for the larger particle sizes.

Electrospun Membrane Surface Modification by Sonocoating with HA and ZnO:Ag Nanoparticles-Characterization and Evaluation of Osteoblasts and Bacterial Cell Behavior In Vitro.

Guided tissue regeneration and guided bone regeneration membranes are some of the most common products used for bone regeneration in periodontal dentistry. The main disadvantage of commercially available membranes is their lack of bone cell stimulation and easy bacterial colonization. The aim of this work was to design and fabricate a new membrane construct composed of electrospun poly (D,L-lactic acid)/poly (lactic-co-glycolic acid) fibers sonocoated with layers of nanoparticles with specific properties, i.e., hydroxyapatite and bimetallic nanocomposite of zinc oxide-silver. Thus, within this study, four different variants of biomaterials were evaluated, namely: poly (D,L-lactic acid)/poly (lactic-co-glycolic acid) biomaterial, poly(D,L-lactic acid)/poly (lactic-co-glycolic acid)/nano hydroxyapatite biomaterial, poly (D,L-lactic acid)/poly (lactic-co-glycolic acid)/nano zinc oxide-silver biomaterial, and poly (D,L-lactic acid)/poly (lactic-co-glycolic acid)/nano hydroxyapatite/nano zinc oxide-silver biomaterial. First, it was demonstrated that the wettability of biomaterials-a prerequisite property important for ensuring desired biological response-was highly increased after the sonocoating process. Moreover, it was indicated that biomaterials composed of poly (D,L-lactic acid)/poly (lactic-co-glycolic acid) with or without a nano hydroxyapatite layer allowed proper osteoblast growth and proliferation, but did not have antibacterial properties. Addition of a nano zinc oxide-silver layer to the biomaterial inhibited growth of bacterial cells around the membrane, but at the same time induced very high cytotoxicity towards osteoblasts. Most importantly, enrichment of this biomaterial with a supplementary underlayer of nano hydroxyapatite allowed for the preservation of antibacterial properties and also a decrease in the cytotoxicity towards bone cells, associated with the presence of a nano zinc oxide-silver layer. Thus, the final structure of the composite poly (D,L-lactic acid)/poly (lactic-co-glycolic acid)/nano hydroxyapatite/nano zinc oxide-silver seems to be a promising construct for tissue engineering products, especially guided tissue regeneration/guided bone regeneration membranes. Nevertheless, additional research is needed in order to improve the developed construct, which will simultaneously protect the biomaterial from bacterial colonization and enhance the bone regeneration properties.

Microstructure and Mechanical Properties of Inverse Nanocomposite Made from Polylactide and Hydroxyapatite Nanoparticles.

Polymer nanocomposites have been extensively researched for a variety of applications, including medical osteoregenerative implants. However, no satisfactory solution has yet been found for regeneration of big, and so-called critical, bone losses. The requirement is to create a resorbable material which is characterised by optimum porosity, sufficient strength, and elastic modulus matching that of the bone, thus stimulating tissue regrowth. Inverse nanocomposites, where the ceramic content is larger than the polymer content, are a recent development. Due to their high ceramic content, they may offer the required properties for bone implants, currently not met by polymer nanocomposites with a small number of nanoparticles. This paper presents inverse nanocomposites composed of bioresorbable nano crystalline hydroxyapatite (HAP NPs) and polylactide (PLLA), produced by cryomilling and a warm isostatic pressing method. The following compositions were studied: 25%, 50%, and 75% of HAP NPs by volume. The mechanical properties and structure of these composites were examined. It was discovered that 50% volume content was optimal as far as compressive strength and porosity are concerned. The inverse nanocomposite with 50% nanoceramics volume displayed a compressive strength of 99 ± 4 MPa, a contact angle of 50°, and 25% porosity, which make this material a candidate for further studies as a bioresorbable bone implant.

Effective Targeting of Colon Cancer Cells with Piperine Natural Anticancer Prodrug Using Functionalized Clusters of Hydroxyapatite Nanoparticles.

Targeted drug delivery offers great opportunities for treating cancer. Here, we developed a novel anticancer targeted delivery system for piperine (Pip), an alkaloid prodrug derived from black pepper that exhibits anticancer effects. The tailored delivery system comprises aggregated hydroxyapatite nanoparticles (HAPs) functionalized with phosphonate groups (HAP-Ps). Pip was loaded into HAPs and HAP-Ps at pH 7.2 and 9.3 to obtain nanoformulations. The nanoformulations were characterized using several techniques and the release kinetics and anticancer effects investigated in vitro. The Pip loading capacity was >20%. Prolonged release was observed with kinetics dependent on pH, surface modification, and coating. The nanoformulations fully inhibited monolayer HCT116 colon cancer cells compared to Caco2 colon cancer and MCF7 breast cancer cells after 72 h, whereas free Pip had a weaker effect. The nanoformulations inhibited ~60% in HCT116 spheroids compared to free Pip. The Pip-loaded nanoparticles were also coated with gum Arabic and functionalized with folic acid as a targeting ligand. These functionalized nanoformulations had the lowest cytotoxicity towards normal WI-38 fibroblast cells. These preliminary findings suggest that the targeted delivery system comprising HAP aggregates loaded with Pip, coated with gum Arabic, and functionalized with folic acid are a potentially efficient agent against colon cancer.

Polymer Membranes Sonocoated and Electrosprayed with Nano-Hydroxyapatite for Periodontal Tissues Regeneration.

Diseases of periodontal tissues are a considerable clinical problem, connected with inflammatory processes and bone loss. The healing process often requires reconstruction of lost bone in the periodontal area. For that purpose, various membranes are used to prevent ingrowth of epithelium in the tissue defect and enhance bone regeneration. Currently-used membranes are mainly non-resorbable or are derived from animal tissues. Thus, there is an urgent need for non-animal-derived bioresorbable membranes with tuned resorption rates and porosity optimized for the circulation of body nutrients. We demonstrate membranes produced by the electrospinning of biodegradable polymers (PDLLA/PLGA) coated with nanohydroxyapatite (nHA). The nHA coating was made using two methods: sonocoating and electrospraying of nHA suspensions. In a simulated degradation study, for electrosprayed membranes, short-term calcium release was observed, followed by hydrolytic degradation. Sonocoating produced a well-adhering nHA layer with full coverage of the fibers. The layer slowed the polymer degradation and increased the membrane wettability. Due to gradual release of calcium ions the degradation-associated acidity of the polymer was neutralized. The sonocoated membranes exhibited good cellular metabolic activity responses against MG-63 and BJ cells. The collected results suggest their potential use in Guided Tissue Regeneration (GTR) and Guided Bone Regeneration (GBR) periodontal procedures.

Nanoparticle Size Effect on Water Vapour Adsorption by Hydroxyapatite.

Handling and properties of nanoparticles strongly depend on processes that take place on their surface. Specific surface area and adsorption capacity strongly increase as the nanoparticle size decreases. A crucial factor is adsorption of water from ambient atmosphere. Considering the ever-growing number of hydroxyapatite nanoparticles applications, we decided to investigate how the size of nanoparticles and the changes in relative air humidity affect adsorption of water on their surface. Hydroxyapatite nanoparticles of two sizes: 10 and 40 nm, were tested. It was found that the nanoparticle size has a strong effect on the kinetics and efficiency of water adsorption. For the same value of water activity, the quantity of water adsorbed on the surface of 10 nm nano-hydroxyapatite was five times greater than that adsorbed on the 40 nm. Based on the adsorption isotherm fitting method, it was found that a multilayer physical adsorption mechanism was active. The number of adsorbed water layers at constant humidity strongly depends on particles size and reaches even 23 layers for the 10 nm particles. The amount of water adsorbed on these particles was surprisingly high, comparable to the amount of water absorbed by the commonly used moisture-sorbent silica gel.

Influence of hydrothermal synthesis parameters on the properties of hydroxyapatite nanoparticles.

Hydroxyapatite (HAp) nanoparticles of tunable diameter were obtained by the precipitation method at room temperature and by microwave hydrothermal synthesis (MHS). The following parameters of the obtained nanostructured HAp were determined: pycnometric density, specific surface area, phase purity, lattice parameters, particle size, particle size distribution, water content, and structure. HAp nanoparticle morphology and structure were determined using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). X-ray diffraction measurements confirmed crystalline HAp was synthesized, which was pure in terms of phase. It was shown that by changing the synthesis parameters, the diameter of HAp nanoparticles could be controlled. The average diameter of the HAp nanoparticles was determined by Scherrer's equation via the Nanopowder XRD Processor Demo web application, which interprets the results of specific surface area and TEM measurements using the dark-field technique. The obtained nanoparticles with average particle diameter ranging from 8-39 nm were characterized by having homogeneous morphology with a needle shape and a narrow particle size distribution. Strong similarities were found when comparing the properties of some types of nanostructured hydroxyapatite with natural occurring apatite found in animal bones and teeth.