Functionalized Hydroxyapatite Nanoparticles Research Articles

Inclusion of modified nano-magnesium hydroxide as an adjuvant flame retardant in the development of PLA/hydroxyapatite nanocomposites

For the preparation of thermal stable and flame-retardant polylactic acid (PLA) nanocomposites two new modified nano-structures including imide-carboxylated chitosan modified Mg(OH)2 (ICMH) and imide-silane functionalized hydroxyapatite nanoparticles (ISHA) were prepared. The structure, morphology, and properties of ICMH and ISHA were investigated using Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), Field-emission scanning electron microscopy (FE-SEM), and thermogravimetric analysis (TGA). The PLA nanocomposites were prepared via solution casting method. The results of Transmission electron microscopy (TEM) and SEM indicated a homogeneous structure for the PLA nanocomposites. The TGA results revealed that the presence of hydroxyapatite nanoparticles (HA) led to increase the thermal resistance of the PLA nanocomposites containing Mg(OH)2 (MDH). Compared to pure PLA, the Tmax value of the PLA sample containing 3 wt% of each filler (the PMH6 sample) enhanced from 364 °C to 372 °C. The results from the microscale combustion calorimeter (MCC) test illustrated that the key parameter values of the PLA nanocomposites were reduced, as compared to the pure PLA. The peak heat release rate (pHRR) and heat release capacity (HRC) values of PMH6 decreased 27 % and 35 %, respectively, as compared to the pure PLA. Also, flammability of PMH6 was reduced, as compared to the pure PLA, with UL-94 V-1 rating and LOI value 26.8 %. Furthermore, the tensile strength of the mentioned sample increased from 36.04 MPa to 38.58 MPa.

Interaction and mechanism between arginine functionalized hydroxyapatite nanoparticles and human umbilical vein endothelial cells

The interaction between terbium-doped arginine functionalized hydroxyapatite nanoparticles (Arg@HAPTb NPs) and cells, and the hindered endocytosis mechanisms of human umbilical vein endothelial cells (HUVECs) were studied by observing the cell uptake of Arg@HAPTb NPs with laser scanning confocal microscopy. Average fluorescence intensity of cells after uptaking different concentrations of NPs was determined by flow cytometer. The results indicate that internalized Arg@HAPTb NPs mainly exist in the cytoplasm of HUVECs, and most of them distribute around the cell nuclei. The entrapment of Arg@HAPTb NPs shows time-dependent and concentration- dependent manners with optimal time of 4 h and concentration of 50 mg/L. Arginine modification significantly improves the uptake efficiency of HAPTb NPs. The entry of Arg@HAPTb NPs into HUVECs is mainly through concave protein-mediated endocytosis, an energy-dependent active transport process.

Self-fluorescence property of octa-arginine functionalized hydroxyapatite nanoparticles aids in studying their intracellular fate in R1 ESCs

Deciphering the endocytosis mechanisms of nanoparticle entry in cells is crucial to understand the fate of nanoparticles and the biological activity of the transported cargo. Such studies require the use of reporter agents such as fluorescent markers. Previously, we have reported the synthesis of self-fluorescent HAp nanoparticles as efficient nucleic acid delivery agents in prokaryotic and eukaryotic cells. Here, we show the application of biocompatible self-fluorescent nano delivery vehicle based on HAp and CPP- octa-arginine as an efficient system to study the mechanisms of intracellular fate of a gene delivery agent. The pathway of octa-arginine functionalized HAp NP (R8HNP) and HAp NP uptake in R1 ESCs was elucidated using confocal microscopy with the help of endocytic inhibitors. The NPs mainly enter R1 ESCs by clathrin mediated and macropinocytosis pathways. It was established that the NPs escape endosomal degradation by proton sponge effect owing to their ability to buffer the pH and trigger osmotic rupture. The functionalization of CPP, effectively enhanced the internalization and endosomal escape in R1 ESCs. The detailed results of these studies are outlined in this manuscript.

Design and development of biomimetic electrospun sulphonated polyether ether ketone nanofibrous scaffold for bone tissue regeneration applications: in vitro and in vivo study

Bone defect restoration remains challenging in orthopedic medical practices. In this study an attempt is carried out to probe the use of new biomimetic SPEEK (sulfonated polyether ether ketone) based nanofibrous scaffold to deliver amine functionalized hydroxyapatite nanoparticles loaded resveratrol for its potent functionality in osteogenic differentiation. SPEEK polymer with reactive functional group SO3H was synthesized through process of sulphonation reaction. Amine functionalized nanoparticles with protonated amino groups revamp the molecular interaction by the formation of hydrogen bonds that in turn intensify the bioactivity of the nanofibrous scaffold. Osteoconductive functionalized nanohydroxyapatite enhances the cell proliferation and osteogenicity with improved cell attachment and spreading. The results of FT-IR, XRD, Carbon-Silica NMR and EDX analysis confirmed the amine functionalization of the hydroxyapatite nanoparticles. Surface morphological analysis of the fabricated nanofibers through SEM and AFM analysis shows vastly interconnected porous structure that mimics the bone extracellular matrix, which enhances the cell compatibility. Cell adhesion and live dead assay of the nanoscaffolds express less cytotoxicity. Mineralization and alkaline phosphatase assay establish the osteogenic differentiation of the nanofibrous scaffold. The in vitro biocompatibility studies reveal that the fabricated scaffold was osteo-compatible with MG63 cell lines. Hemocompatibility study further proved that the designed biomimetic nanofibrous scaffold was highly suitable for bone tissue engineering. The results of in vivo analysis in zebrafish model for the fabricated nanofibers demonstrated significant increase in the caudal fin regeneration indicating mineralization of osteoblast. Thus, the commending results obtained instigate the potentiality of the composite nanofibrous scaffold as an effective biomimetic substrate for bone tissue regeneration.

Modulating response of sunflower (Hellianthus annuus) to induced salinity stress through application of engineered urea functionalized hydroxyapatite nanoparticles.

Agro-nanotechnology aims to improve the quality and quantity of plants and plant products while preserving environmental health. Contemporary anecdotal studies that provide representation of the use of nanostructures as fertilizers, pesticides, and plant growth regulators have highlighted the need to determine the effect of such modified nanofertilizers on transforming plant yield under abiotic stress. Present study was performed to modulate the physiological response of Hellianthus annuus through the application of Urea capped hydroxyapatite nanoparticles (Urea-HANPs) in stressed environment. Hydroxyapatite nanoparticles were synthesized via co-precipitation method, functionalized with urea and characterized through a series of contemporary techniques of transmission electron microscopy (TEM), energy dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD), and Fourier transform infrared (FTIR) spectroscopy. We observed that Urea-HANPs significantly (p < .05) ameliorated resistivity in plant to osmotic stress by enhancing agronomic and physiobiochemical attributes. Elevated chlorophyll contents were reported from tested leaves treated with Urea-HANPs in T6 (0.05 M NaCl + 10 μg/ml Urea-HANP) under induced salinity stress. Data revealed significant decrease in osmolytes at T3 (0.1M NaCl), and T4 (0.2M NaCl) which was significantly ameliorated in T9 (0.1M NaCl + 10μg/ml Urea-HANPs) and T12 (0.2M NaCl + 10μg/ml Urea-HANPs). The antioxidant system was boosted up by the application of Urea-HANPs preventing the plant from oxidative stress by scavenging reactive oxygen species (ROS). It has been concluded that alleviation in impact of osmotic stress on plant through the use of Urea-HANPs was coupled with elevation in photosynthetic performance, stimulation of osmolytes and boosting antioxidant system of plants.

Hydroxyapatite reinforced inorganic-organic hybrid nanocomposite as high-performance adsorbents for bilirubin removal in vitro and in pig models

Highly efficient removal of bilirubin from whole blood directly by hemoperfusion for liver failure therapy remains a challenge in the clinical field due to the low adsorption capacity, poor mechanical strength and low biocompatibility of adsorbents. In this work, a new class of nanocomposite adsorbents was constructed through an inorganic-organic co-crosslinked nanocomposite network between vinyltriethoxysilane (VTES)-functionalized hydroxyapatite nanoparticles (V-Hap) and non-ionic styrene-divinylbenzene (PS-DVB) resins (PS-DVB/V-Hap) using suspension polymerization. Notably, our adsorbent demonstrated substantially improved mechanical performance compared to the pure polymer, with the hardness and modulus increasing by nearly 3 and 2.5 times, respectively. Moreover, due to the development of a mesoporous structure, the prepared PS-DVB/V-Hap3 exhibited an ideal adsorption capacity of 40.27 mg g−1. More importantly, the obtained adsorbent beads showed outstanding blood compatibility and biocompatibility. Furthermore, in vivo extracorporeal hemoperfusion verified the efficacy and biosafety of the adsorbent for directly removing bilirubin from whole blood in pig models, and this material could potentially prevent liver damage and improve clinical outcomes. Taken together, the results suggest that PS-DVB/V-Hap3 beads can be used in commercial adsorption columns to threat hyperbilirubinemia patients through hemoperfusion, thus replacing the existing techniques where plasma separation is initially required.

Design of a Biocompatible Hydroxyapatite-Based Nanovehicle for Efficient Delivery of Small Interference Ribonucleic Acid into Mouse Embryonic Stem Cells.

The small interference RNA (siRNA)-assisted RNA interference approach in stem cells for differentiating into cell-specific lineages is gaining importance for its therapeutic potential. An effective gene delivery platform is crucial to achieve this goal. In this context, self-fluorescent, cell-penetrating peptide (CPP)-functionalized hydroxyapatite nanoparticles (R8HNPs) were synthesized by a modified sol gel technique. R8HNPs were crystalline, displayed characteristic bands, and exhibited broad emission spectra from 350 to 750 nm corresponding to green and red fluorescence. The biocompatible R8HNPs displayed robust binding with siRNA and excellent uptake in R1 ESCs. This was attributed to functionalization with CPP. Moreover, the R8HNP-complexed siRNA exhibited excellent serum and room temperature stability. The NPs protected the siRNA from sonication, pH, and temperature-induced stress and efficiently delivered siRNA to trigger 80% silencing of a pluripotency marker gene, Oct4, in R1 ESCs at 48 h. The transient downregulation was also observed at the protein level. Our findings demonstrate R8HNPs as a promising delivery agent for siRNA therapeutics with the potential for lineage-specific differentiation and future applications in regenerative medicine.

Structural and Functional Stability of MRSA Membrane Proteins on Interaction with Functionalized Hydroxyapatite Nanoparticles: A Target Mechanism to Combat Multi Drug Resistant Bacteria

Structural and Functional Stability of MRSA Membrane Proteins on Interaction with Functionalized Hydroxyapatite Nanoparticles: A Target Mechanism to Combat Multi Drug Resistant Bacteria

Antibacterial potential of nano-particulate intracanal medications on a mature E. faecalis biofilm in an ex-vivo model

Aim: To investigate and compare the antibacterial and antibiofilm activity of Chlorhexidine Digluconate functionalized Hydroxyapatite Nanoparticles and silver Nanoparticles as intracanal medicaments on a mature E. faecalis biofilm model in root canals of extracted teeth. Methodology: Sixty-eight human maxillary central incisors were mechanically prepared, sterilized, infected with E. faecalis, and then incubated for 28 days under anaerobic conditions to develop a mature E. faecalis biofilm. Eight teeth were used to monitor biofilm formation and maturation over the incubation period with field emission scanning electron microscopy (Fe SEM), while the other 60 teeth were divided into two experimental groups (n=20) in which 2% chlorhexidine digluconate functionalized hydroxyapatite nanoparticles (Group A) and 0.02% silver nanoparticles (group B) were used as intra canal medications, and two control groups (n=10). The positive control group (Group C) was used to check for bacterial viability throughout the experiment, while the negative control group (Group D) was used to check for sterility of the procedures. Finally, bacterial samples were collected and analyzed quantitively by culture counts and qualitatively by real time PCR (RTQ-PCR). Results: RTQ-PCR detected E. faecalis DNA in all groups except for the negative control group. The mean values of E. faecalis DNA detected in groups A, B and C were 99.6 ng/mL, 67.1 ng/mL , 2797.4 ng/mL respectively. Statistical analysis of these results showed that both treatment groups presented statistically significantly lower mean values of E. faecalis DNA quantities compared to the positive control group. However, there was no statistically significant difference between them (P<.05). Conclusions: Chlorhexidine Digluconate functionalized Hydroxyapatite Nanoparticles and silver nanoparticles demonstrated an effective antimicrobial activity against mature E. faecalis biofilms.

Photocrosslinkable nanocomposite ink for printing strong, biodegradable and bioactive bone graft

3D printing is known as a cost-effective technique that shows huge potential in fabrication of graft substitutes for bone tissue regeneration. However, the tradeoff between 3D printability, mechanical strength and bioactivity of the printed materials (i.e., inks) remains a challenge. In this work, we present a novel photocrosslinkable nanocomposite ink composed of tri-block poly (lactide-co-propylene glycol-co-lactide) dimethacrylate (PmLnDMA, m and n respectively represent the unit length of propylene glycol and lactide) and hydroxyethyl methacrylate (HEMA)-functionalized hydroxyapatite nanoparticles (nHAMA). The reactive HEMA-conjugated nHAMA, is designed to covalently crosslink with the surrounding polymer matrix to further increase the interfacial bonding between them. We find that the nHAMA can rapidly interact with PmLnDMA upon light exposure within 140 s and form an inorganic-organic co-crosslinked nanocomposite network, further enhancing the nanofiller-matrix interfacial compatibility. Notably, our nanocomposites possess significantly improved mechanical performances compared to the polymer, with compressive modulus increasing by nearly 10 times (from ⁓40 to ⁓400 MPa). Moreover, thanks to the low exothermic heat generation (<37 °C) during photocrosslinking, our nanocomposite ink enables facile encapsulation and long-term release of heat-labile biomolecules like bone morphogenic protein-2 (BMP-2). Furthermore, it demonstrates a readily tunable rheological property, wettability, degradation, and printability as a 3D bone scaffold. Together with its superior osteogenic ability both in vitro and in vivo, we envision that our nanocomposite ink holds great promise in 3D printing of bone grafts.

Development of surface functionalized hydroxyapatite nanoparticles for enhanced specificity towards tumor cells

Nanoparticles coupled with targeting moieties have attracted a great deal of attention for cancer therapy since they can facilitate site-specific delivery of drug and significantly limit the side effects of systemic chemotherapy. In this study, our aim is to develop surface functionalized hydroxyapatite nanoparticles, which could provide binding sites for a cancer cell targeting ligand, folic acid (FA) as well as an anticancer drug, doxorubicin hydrochloride (DOX). In order to attain dual functionalities, hydroxyapatite nanoparticles were functionalized with gelatin molecules. Gelatin, being a protein has both carboxyl and amine moieties, which makes it suitable for binding of DOX and FA. FA was chemically conjugated to the nanoparticles through an EDCNHS coupling reaction. The formation of single-phase hydroxyapatite nanostructure was ascertained by X-ray diffraction studies and the presence of organic moieties on the surface of nanoparticles was evident from Fourier transform infrared spectroscopy, thermogravimetric analysis and U.V.-visible spectroscopy. The FA-conjugated nanoparticles (FA-Gel-HANPs) showed high affinity towards DOX and pH-responsive sustained release of drug with higher release rate under acidic pH conditions, desired for cancer therapy. The FA-Gel-HANPs showed negligible cytotoxicity towards different cell lines (HepG2, WEHI-164, KB, WI-26 VA4 and WRL-68). However, DOX loaded nanoparticles (DOX-FA-Gel-HANPs) exhibited significant toxicity towards these cells, which was however highest in folate receptor (FR)-overexpressing, KB cells. These results were correlated with enhanced cellular uptake of DOX-FA-Gel-HANPs in KB cells in comparison to FR-deficient, WRL-68 cells studied by confocal laser scanning microscopy and flow cytometry. Moreover, cell cycle analysis in KB cells, showed higher sub-G1 population, indicating apoptosis as one of the cell death mechanisms. Overall, this study suggests that DOX-FA-Gel-HANPs could serve as a promising tumor-targeted drug delivery system.

A novel method for genetic transformation of C. albicans using modified-hydroxyapatite nanoparticles as a plasmid DNA vehicle.

In modern biological research, genetic transformation is an important molecular biology technique with extensive applications. In this work, we describe a new method for the delivery of plasmid DNA (pDNA) into a yeast species, Candida albicans. This method is based on the use of novel arginine–glucose–PEG functionalized hydroxyapatite nanoparticles (M-HAp NPs) as a vehicle which delivers pDNA into Candida albicans with a high transformation efficiency of 106 cfu μg−1 of pDNA, without the need for preparation of competent cells. A four-fold higher transformation efficiency as compared to that of the electroporation method was obtained. This new method could provide exciting opportunities for the advancement of the applications of yeasts in the field of biotechnology.

Catechin-conjugated mesoporous hydroxyapatite nanoparticle: A novel nano-antioxidant with enhanced osteogenic property.

Hydroxyapatite is the main component of mineral phase of bone which is widely employed for coating metal implants and scaffold materials in synthetic bone grafts owing to its osteoinductive property. In order to improve the bioactivity of hydroxyapatite, mesoporous hydroxyapatite nanoparticles (MHAP) were synthesized and chemically functionalized with 3-aminopropyltriethoxysilane. The amine-functionalized nanoparticles were conjugated with a natural antioxidant, catechin (Cat), through a stable amide linkage. The true structure of the bioconstruct was confirmed by calculating condensed Fukui indices. The functionalized-hydroxyapatite nanoparticles (Cat@MHAP) showed an outstanding antioxidant activity, having reactivity toward hydroxyl and superoxide radicals larger than that of free catechin. To explore the bone cell responses to this material, multilayer nanoparticle films were prepared by MHAP and Cat@MHAP on a glass substrate. Afterward, the short- and long-term responses of cultured mesenchymal stem cells (MSCs), osteosarcoma cells (Saos-2), and doxorubicin-resistant cells (RSaos-2/Dox) on the surface of the prepared films were investigated. Both the MSCs and bone tumor cells selectively adhered onto Cat@MHAP surface as compared with glass and MHAP at initial culture time. Moreover, it was found that Cat@MHAP decreases the proliferation of Saos-2 and RSaos-2/Dox cells in a time-dependent manner, while it supports the growth of MSCs, indicating the ability of Cat@MHAP to distinguish tumor cells from normal ones. Further, Cat@MHAP promotes the osteogenic differentiation in both the MSCs and tumor cells, accompanied by the attenuation of intracellular ROS. From these results, Cat@MHAP is a novel "nano-antioxidant," which could be considered as a promising biomaterial in treating bone defects, particularly after surgery in osteosarcoma patients.

Covalent immobilization of doxorubicin in glycine functionalized hydroxyapatite nanoparticles for pH-responsive release

Development and therapeutic evaluation of glycine functionalized hydroxyapatite nanoparticles having a covalently conjugated anticancer drug, doxorubicin hydrochloride.

XRD, Thermal, Haemolysis and DNA Binding Studies of L-Arginine Functionalized Hydroxyapatite Nano-particles

Hydroxyapatite (HAP), an important bio-mineral, finds numerous applications in the clinical field. As amino acids exhibit synergetic effects on structure, morphology and several bio-logical properties of HAP, an attempt was made by the present authors to synthesize pure and different amount of L-arginine functionalized HAP nano-particles by the surfactant mediated approach. The samples were characterized by using different techniques, viz., powder X-Ray Diffraction (powder XRD), Fourier Transform Infrared (FT-IR) spectroscopy, Transmission Electron Microscopy (TEM) and Thermo-Gravimetry Analysis (TGA). The Williamson and Hall (W-H) analysis was performed on the powder XRD patterns, which indicated the particle size in nano-meter range and the strain associated with nano- particles changed with functionalization of HAP by L-arginine. FTIR study indicated the presence of various functional groups due to the functionalization of L-arginine with HAP. TGA results indicated slightly poor thermal stability of L-arginine functionalized HAP samples with comparison to pure HAP. The DNA binding studies indicated improved DNA binding on L-arginine functionalization of HAP. The haemolysis studies were also carried out.

Arginine functionalized hydroxyapatite nanoparticles and its bioactivity for gene delivery

In order to further improve the transfection efficiency of hydroxyapatite nanoparticle (HAp), arginine functionalized hydroxyapatite (HAp/Arg) was synthesized by hydrothermal synthesis. The morphology, crystallite size and zeta potential of the HAp/Arg were characterized by transmission electron microscopy (TEM), atomic force microscopy (AFM) and zeta potential analyzer. The loading and protecting properties of HAp/Arg to DNA were tested by electrophoresis. Its cytotoxicity was also measured in Hela cells and HAEC cells by MTT and LDH, and its transfection efficiency was examined by fluorescence microscope and flow cytometry. The results reveal that HAp/Arg is short rod-like and nano single crystal, the mean diameter is 50–90 nm and zeta potential is 35.8 mV at pH 7.4. HAp/Arg to DNA can be condensed by electrostatic effect and protect DNA against degradation in DNase I, and shows high transfection efficiency without cytotoxicity. These results suggest that HAp/Arg can be a promising alternative as a novel gene delivery system.

Surface modification and paclitaxel drug delivery of folic acid modified polyethylene glycol functionalized hydroxyapatite nanoparticles

Abstract Nanoparticles are used for a number of biomedical applications. In this work we report the synthesis of folic acid (FA) modified polyethylene glycol (PEG) functionalized hydroxyapatite (HAp) nanoparticles. The anticancer drug, paclitaxel, is attached to the folic acid modified polyethylene glycol functionalized hydroxyapatite nanoparticles and the in vitro drug release is analyzed. The surface modification and functionalization is confirmed by Fourier transform infrared spectroscopy (FTIR), thermo gravimetric analysis (TGA) and UV spectroscopy. The importance of the paper is the investigation of the release behavior of paclitaxel conjugated folic acid modified polyethylene glycol functionalized hydroxyapatite nanoparticles. The results show an initial rapid release and then a sustained release.