nHA Particles Research Articles

Enhancing Bioactivity and Mechanical Properties of Nano-Hydroxyapatite Derived from Oyster Shells through Hydrothermal Synthesis.

Nano-hydroxyapatite (nHA) demonstrates favorable biological activity, cell adhesion, cell proliferation, and osteoconductivity, making it highly valuable in biomedicine. It is extensively used as a bone substitute and in bone transplantation within the dental and orthopedic fields. This study employed oyster shells as a calcium source to synthesize nHA at 150 °C with various hydrothermal reaction durations (10 min, 1 h, 6 h, and 12 h). As a control, HA synthesized via a wet precipitation method for 1 h at room temperature was utilized. Subsequent material analyses, including XRD, FE-SEM, FTIR, and ICP-MS, were conducted, followed by comprehensive evaluations of the bioactivity, cell attachment, cell proliferation, and sintering properties of the synthesized nHA. The results indicated that nHA synthesized through the hydrothermal reaction produced nanoscale crystals, with the aspect ratio of nHA particles increasing with the duration of hydrothermal treatment. Notably, rod-like nHA particles became prominent with hydrothermal durations exceeding 6 h. nHA particles derived from oyster shells contained carbonate and trace elements (Na, Mg, K, and Sr), similar to constituents found in human hard tissue such as bone and teeth. The immersion of nHA synthesized at 150 °C for 1 h (HT2) in simulated body fluid (SBF) for 28 d led to the formation of a bone-like apatite layer on the surface, indicating the excellent bioactivity of the synthesized nHA. The cell culture results revealed superior cell attachment and proliferation for nHA (HT2). Following the sequential formation and sintering at 1200 °C for 4 h, HT2 ceramics exhibited enhanced microhardness (5.65 GPa) and fracture toughness (1.23 MPa·m0.5), surpassing those of human tooth enamel.

Centrifugally spun hydroxyapatite/carbon composite nanofiber scaffolds for bone tissue engineering

In recent years, advancements in tissue engineering have demonstrated the potential to expedite bone matrix formation, leading to shorter recovery times and decreased clinical challenges compared to conventional methods. Therefore, this study aims to develop composite carbon nanofibers (CNFs) integrated with nano-hydroxyapatite (nHA) particles as scaffolds for bone tissue engineering applications. A key strategy in achieving this objective involves harnessing nanofibrous structures, which offer a high surface area, coupled with nHA particles expected to accelerate bone regeneration and enhance biological activity. To realize this, polyacrylonitrile (PAN)/nHA nanofibers were fabricated using the centrifugal spinning (C-Spin) technique and subsequently carbonized to yield CNF/nHA composite structures. Scanning Electron Microscopy (SEM) confirmed C-Spin as a suitable method for PAN and CNF nanofiber production, with nHA particles uniformly dispersed throughout the nanofibrous structure. Carbonization resulted in reduced fiber diameter due to thermal decomposition and shrinkage of PAN molecules during the process. Furthermore, the incorporation of nHA particles into PAN lowered the stabilization temperature (by 5 °C–20 °C). Tensile tests revealed that PAN samples experienced an approximately 80% increase in ultimate tensile strength and a 187% increase in modulus with a 5 wt.% nHA loading. However, following carbonization, CNF samples exhibited a 50% decrease in strength compared to PAN samples. Additionally, the addition of nHA into CNF improved the graphitic structure. The incorporation of nHA particles into the spinning solution represents a viable strategy for enhancing CNF bioactivity.

Synthesis and Viscoelastic Properties of Polycaprolactone/Polyvinylidene Fluoride/Nanohydroxyapatite Composite Scaffolds

AbstractObtaining a polymer nanocomposite with optimum viscoelastic, thermal, and biocompatibility properties is the main objective when designing nanocomposite systems with potential applications in tissue engineering. For this purpose, a blend of Polycaprolactone (PCL) and Polyvinylidene fluoride (PVDF) in an 85/15 weight ratio, along with a nanocomposite reinforced by nanohydroxyapatite (nHA) particles, is fabricated using a solution casting method in a mold. The impact of nHA content on crystallinity, viscoelastic properties, thermal stability, and the properties–structure relationship of nanocomposites is evaluated using scanning electron microscopy (SEM). Dynamic mechanical thermal (DMTA) analysis is used to determine the William–Landel–Ferry (WLF) constants and the effect of nHA on the nanocomposite's viscoelastic behavior. The PCL/15PVDF/0.5 wt% nHA exhibits the maximum thermal stability (40% residual char value) and 95% increase in storage modulus at 90 °C (rubbery region) in comparison with PCL/15PVDF blend. Water contact angle (WCA) and biocompatibility tests are conducted on the PCL/15PVDF blend and nanocomposite scaffolds to design appropriate nanocomposite systems with potential applications in tissue engineering. The high hydrophilic properties are assigned to PCL/15PVDF/0.5 wt% nHA with a WCA of 67.5°. Finally, in vitro cell culture confirmed 0.5 wt% nHA significantly improves cell adhesion and cytotoxicity with MG‐63 cells.

Dispersion Strategy improves the mechanical properties of 3D-Printed biopolymer nanocomposite

Homogenous dispersion of nanoparticles in polymer matrices is a technical challenge that if overcome can lead to improved mechanical properties of the resulting nanocomposites. In this work, we successfully refined commercially-available nanoscale, calcium deficient, and poorly crystalline hydroxyapatite (nHA) particles and composited them with acrylate and methacrylate functionalized soybean oil (mAESO) and triethylene glycol dimethacrylate (TEGDMA) producing inks for masked stereolithography (mSLA) -based 3D printing. First, we used shear mixing and ultrasonication on nHA/ethanol mixtures to break down agglomerates and then separated the finest nanoparticles from the remaining agglomerates using centrifugation. The refined nanoparticles (termed fine) were then mixed with the resins and UV-initiator to produce inks for 3D printing. Similarly, we prepared one ink using as-purchased nHA particles (termed raw) and another ink using leftover agglomerates after refinement (termed coarse). We compared the rheological properties of the nHA-resin inks. We used mSLA to fabricate nanocomposite specimens and tested them using flexural, and Mode-I fracture toughness testing following ASTM standards. The dispersion of nanoparticles in the polymer matrix was studied by analyzing backscattered mode scanning electron microscopy images. The nHA particle refinement improved the nanoparticle dispersion in the resin matrix while also increasing the viscosity and shear yield strength of the nanocomposite ink. The flexural fracture strength, flexural modulus, and Mode-I fracture toughness of refined nHA-based nanocomposites were increased by 11 %, 71 %, and 12 %, respectively compared to the raw nHA-based nanocomposites. However, the flexural fracture strain of refined nHA-based nanocomposites was lower by 40 % compared to the raw nHA-based nanocomposites. The nanocomposites became stiffer with the incorporation of refined nanoscale nHA. The separation of nanoscale nHA particles, excellent dispersion of these nanoparticles in polymer matrix, and improved flexural strength and modulus opens a new avenue towards the 3D printing of high-performance nHA-based nanocomposites.

Functionalization of calcium-deficient nanohydroxyapatite improves the mechanical properties of 3D printed biopolymer nanocomposites

Agglomerations of nanoparticles in a polymer matrix can drastically reduce the mechanical properties of a polymer nanocomposite, especially its strength. The grafting of nanoparticle surfaces with suitable functional groups can provide improved dispersion and stronger interfacial bonding, improving the fracture resistance of the nanocomposite. In this study, calcium-deficient nanohydroxyapatite (nHA) particles were functionalized with an amino acid-based urethane methacrylate (lysine urethane methacrylate, LUM) and subsequently reacted with hydroxyethyl methacrylate. We mixed these functionalized nHA particles with resin, composed of methacrylated acrylated epoxidized soybean oil, methacrylated isosorbide, and triethylene glycol dimethacrylate, and 3D-printed nanocomposites using masked stereolithography. We hypothesized that the functionalized nanoparticles would enhance the mechanical performance of the 3D-printed nanocomposites due to the greater dispersion and stronger interface. Flexural, tensile, compression and Mode-I fracture toughness test specimens were fabricated using a mSLA printer and tested following ASTM standards. The LUM functionalization of nHA improved the dispersion and increased the viscosity of the uncured nanocomposite ink. The flexural fracture strength, yield strength, and mode-I fracture toughness values were increased by 10 %, 30 %, and 11 %, respectively. The LUM improved the strength and fracture toughness by providing a stronger, more stable interface, resisting debonding between the matrix and particles, allowing for greater plastic deformation.

Influence of size and crystallinity of nanohydroxyapatite (nHA) particles on the properties of Polylactic Acid/nHA nanocomposite scaffolds produced by 3D printing

Polylactic acid (PLA) and hydroxyapatite (HA) composite scaffolds have been widely studied for applications in bone tissue engineering (BTE) due to their bioactive and biocompatible properties. However, there is a need for more knowledge about the influence of size and crystallinity of HA nanoparticles (nHA) on the properties of PLA/nHA nanocomposite scaffolds produced by 3D printing. In this study, 3D printing was used to produce PLA nanocomposite scaffolds incorporated with nHA filler with different particle sizes and crystallinities. Initially, the nanocomposites were prepared by casting for 3D printing of scaffolds, which were characterized by analysis of thermal, morphological, physical-chemical, mechanical, and biological properties in vitro. The results showed that the size and crystallinity of nHA particles mainly influenced the scaffolds' mechanical properties, degradation rate, and bioactivity. Incorporating nHA provided a gain in compressive strength compared to pure PLA, superior to natural cancellous bone, which varies between 2 and 12 MPa. The lower crystallinity, 39.46 %, promoted a higher rate of degradation and bioactivity in vitro due to its solubility in the simulated fluid. All nanocomposite scaffolds showed cell viability above 90 %. The scaffolds showed suitable properties for BTE applications.

Effects of friction stir processing and nano-hydroxyapatite on the microstructure, hardness, degradation rate and in-vitro bioactivity of WE43 alloy for biomedical applications

Nowadays, magnesium alloys are emerging in biomedical implants for their similar properties to natural bones. However, the rapid degradation of magnesium alloys in biological media hinders successful implantation. Refinement of microstructure, as well as reinforcement particles can significantly improve the degradation rate. In this work, multi-pass friction stir processing (FSP) was proposed to synthesize WE43/nano-hydroxyapatite (nHA) surface composite, the microstructure, reinforced particle distribution, micro-hardness, corrosion behavior and in-vitro bioactivity were studied. The subsequent FSP passes of WE43 alloy and WE43/nHA composite refined the grain size which was reduced by 94.29% and 95.92% (2.63 and 1.88 µm, respectively) compared to base metal after three passes. This resulted in increasing the microhardness by 120% (90.86 HV0.1) and 135% (105.59 HV0.1) for the WE43 and WE43-nHA, respectively. It is found that increasing FSP passes improved the uniform distribution of nHA particles within the composite matrix which led to improved corrosion resistance and less degradation rate. The corrosion rate of the FSPed WE43/nHA composite after three passes was reduced by 38.2% (4.13 mm/year) and the degradation rate was reduced by 69.7% (2.87 mm/y). This is attributed to secondary phase (Mg24Y5 and Mg41Nd5) particle fragmentation and redistribution, as well as a homogeneous distribution of nHA. Additionally, the growing Ca-P and Mg(OH)2 layer formed on the surface represented a protective layer that reduced the degradation rate. The wettability test revealed a relatively hydrophilic surface with water contact angle of 49.1 ± 2.2° compared to 71.2 ± 2.1° for base metal. Also, biomineralization test showed that apatite layer grew after immersion 7d in simulated body fluid with atomic ratio of Ca/P 1.60 approaching the stoichiometric ratio (1.67) indicating superior bioactivity of FSPed WE43/nHA composite after three passes. These results raise that the grain refinement by FSP and introduction of nHA particles significantly improved the degradation rate and in-vitro bioactivity of WE43 alloy for biomedical applications.

Bio-Based PLA/PBS/PBAT Ternary Blends with Added Nanohydroxyapatite: A Thermal, Physical, and Mechanical Study.

The physical and mechanical properties of novel bio-based polymer blends of polylactic acid (PLA), poly(butylene succinate) (PBS), and poly (butylene adipate-co-terephthalate) (PBAT) with various added amounts of nanohydroxyapatite (nHA) were investigated in this study. The formulations of PLA/PBS/PBAT/nHA blends were divided into two series, A and B, containing 70 or 80 wt% PLA, respectively. Samples of four specimens per series were prepared using a twin-screw extruder, and different amounts of nHA were added to meet the regeneration needs of bone graft materials. FTIR and XRD analyses were employed to identify the presence of each polymer and nHA in the various blends. The crystallization behavior of these blends was examined using DSC. Tensile and impact strength tests were performed on all samples to screen feasible formulations of polymer blends for bone graft material applications. Surface morphology analyses were conducted using SEM, and the dispersion of nHA particles in the blends was further tested using TEM. The added nHA also served as a nucleating agent aimed at improving the crystallinity and mechanical properties of the blends. Through the above analyses, the physical and mechanical properties of the polymer blends are reported and the most promising bone graft material formulations are suggested. All blends were tested for thermal degradation analysis using TGA and thermal stability was confirmed. The water absorption experiments carried out in this study showed that the addition of nHA could improve the hydrophilicity of the blends.

Polyether ether ketone coated with nanohydroxyapatite/graphene oxide composite promotes bioactivity and antibacterial activity at the surface of the material.

Polyether ether ketone (PEEK) is considered an alternative material for manufacturing dental implants. However, PEEK lacks bioactivity and antibacterial action. In a series of experiments designed to enhance the surface properties of PEEK, we present a nanohydroxyapatite (nHA) and graphene oxide (GO) composite as a coating for PEEK-based dental implants to improve biological properties and antibacterial action. PEEK discs were polished, cleaned, and coated with the composite consisting of nHA particles doped with 0.75wt% graphene oxide by a micro-emulsion technique according to patent US8,206,813. X-ray diffraction, field emission scanning electron microscopy-energy dispersive spectroscopy, and atomic force microscopy were utilized to characterize the composite coating. The wettability of the coated and non-coated samples was assessed by optical contact angle measurement. Antibacterial action of the composite coating was explored against S. aureus and E. coli and cytotoxicity determined utilizing osteoblast-like cells and gingival fibroblasts. The findings showed that the nHA/GO composite coating, approximately 1.3 μm thick, was homogenous with few micro-cracks and adhered to the PEEK surface. The surface roughness was reduced to 21.26nm and the wettability was improved to 54.6⁰ after coating with the composite coating. Antibacterial activity was moderate, killing 99% of S. aureus and E. coli, with acceptable levels of cytotoxicity to mammalian osteoblast-like cells and gingival fibroblasts.

Effects of biopolymer functionalization and nanohydroxyapatite heat treatment on the tensile and thermomechanical properties of Bone-Inspired 3D printable nanocomposite biomaterials

Bone-inspired biopolymer nanocomposite grafts are an alternative to conventional bone substitutes if designed to offer a structure with adequate mechanical properties and biocompatibility. In this study, a set of novel 3D printable bone-inspired nanocomposite biomaterials was developed to investigate the effects of additional cross-linking of the biopolymer matrix and heat treatment of nHA particles on the tensile and thermo-mechanical properties of these nanocomposites used in extrusion-based 3D printing. We observed that additional functionalization of acrylated epoxidized soybean oil (AESO), as the main component of the biopolymer matrix, with additional methacrylate groups (mAESO), increased the strength and elastic modulus of extruded nanocomposites by more than three times, as well as doubled the glass transition temperature due to an increased degree of crosslinking in the functionalized matrices. The mAESO-based nanocomposite filaments demonstrated a strength safety factor of 1.5 against the minimum required standard mechanical properties for bone cement according to ISO standard5833. While heat treatment of nHA reduced the frequency of larger agglomerations, no significant difference in mechanical properties was observed. These novel 3D printable nanocomposite biomaterials with their improved strength, modulus and thermo-mechanical properties could be suitable candidates for fabricating complex ‘by design’ 3D printed grafts and scaffolds for bone reconstruction.

Longitudinal in vivo biodistribution of nano and micro sized hydroxyapatite particles implanted in a bone defect.

Hydroxyapatite (HA) has been widely used as a bone substitute and more recently as a carrier for local delivery of bone targeted drugs. Majority of the approved HA based biomaterials and drug carriers comprise of micrometer sized particulate HA (mHA) or granules and can therefore only be used for extracellular drug release. This shortcoming could be overcome with the use of cell penetrating HA nanoparticles (nHA) but a major concern with the clinical use of nHA is the lack of data on its in vivo biodistribution after implantation. In this study, we aimed to study the in vivo biodistribution of locally implanted nHA in a clinically relevant tibial void in rats and compare it with mHA or a combination of mHA and nHA. To enable in vivo tracking, HA particles were first labelled with 14C-zoledronic acid (14C-ZA), known to have a high binding affinity to HA. The labelled particles were then implanted in the animals and the radioactivity in the proximal tibia and vital organs was detected at various time points (Day 1, 7 and 28) post-implantation using scintillation counting. The local distribution of the particles in the bone was studied with micro-CT. We found that majority (>99.9%) of the implanted HA particles, irrespective of the size, stayed locally at the implantation site even after 28days and the findings were confirmed using micro-CT. Less than 0.1% radioactivity was observed in the kidney and the spleen at later time points of day 7 and 28. No pathological changes in any of the vital organs could be observed histologically. This is the first longitudinal in vivo HA biodistribution study showing that the local implantation of nHA particles in bone is safe and that nHA could potentially be used for localized drug delivery.

Valorization of fish and shrimp wastes to nano-hydroxyapatite/chitosan biocomposite for wastewater treatment

The conversion of the fish and shrimp wastes to produce value-added products brings economic benefits and reduces the risk of environmental pollution. This study presents a facile method for preparing nano-hydroxyapatite/chitosan biocomposites (nHCB) for wastewater treatment and bacterial immobilization. It is a novelty that hydroxyapatite nanoparticles (nHA) and chitosan (CTS) were recovered from fishbone and shrimp shell sources. The prepared nHCBs showed a high porosity, and nHA particles were well distributed in the CTS matrix. Due to these unique properties as an adsorbent for removing dyes and Cu(II) ions from wastewater, nHCBs exhibited rapid adsorption, high capacity, and easy separation for cyclic use. The maximum sorption capacity were 44.05 mg/g (methylene orange), 84.89 mg/g (methylene blue), and 236.97 mg/g (Cu(II) ions). The findings suggest a great potential fabrication of low-cost bio-composites and their application for wastewater treatment.

Fabrication and Testing of Multi-Hierarchical Porous Scaffolds Designed for Bone Regeneration via Additive Manufacturing Processes.

Bone implants or replacements are very scarce due to the low donor availability and the high rate of body rejection. For this reason, tissue engineering strategies have been developed as alternative solutions to this problem. This research sought to create a cellular scaffold with an intricate and complex network of interconnected pores and microchannels using salt leaching and additive manufacturing (3D printing) methods that mimic the hierarchical internal structure of the bone. A biocompatible hydrogel film (based on poly-ethylene glycol) was used to cover the surface of different polymeric scaffolds. This thin film was then exposed to various stimuli to spontaneously form wrinkled micropatterns, with the aim of increasing the contact area and the material’s biocompatibility. The main innovation of this study was to include these wrinkled micropatterns on the surface of the scaffold by taking advantage of thin polymer film surface instabilities. On the other hand, salt and nano-hydroxyapatite (nHA) particles were included in the polymeric matrix to create a modified filament for 3D printing. The printed part was leached to eliminate porogen particles, leaving homogenously distributed pores on the structure. The pores have a mean size of 26.4 ± 9.9 μm, resulting in a global scaffold porosity of ~42% (including pores and microchannels). The presence of nHA particles, which display a homogeneous distribution according to the FE-SEM and EDX results, have a slight influence on the mechanical resistance of the material, but incredibly, despite being a bioactive compound for bone cells, did not show a significant increase in cell viability on the scaffold surface. However, the synergistic effect between the presence of the hydrogel and the pores on the material does produce an increase in cell viability compared to the control sample and the bare PCL material.

Biomimetic, mussel-inspired surface modification of 3D-printed biodegradable polylactic acid scaffolds with nano-hydroxyapatite for bone tissue engineering.

Polylactic acid (PLA) has been widely used as filaments for material extrusion additive manufacturing (AM) to develop patient-specific scaffolds in bone tissue engineering. Hydroxyapatite (HA), a major component of natural bone, has been extensively recognized as an osteoconductive biomolecule. Here, inspired by the mussel-adhesive phenomenon, in this study, polydopamine (PDA) coating was applied to the surface of 3D printed PLA scaffolds (PLA@PDA), acting as a versatile adhesive platform for immobilizing HA nanoparticles (nHA). Comprehensive analyses were performed to understand the physicochemical properties of the 3D-printed PLA scaffold functionalized with nHA and PDA for their potent clinical application as a bone regenerative substitute. Scanning electron microscopy (SEM) and element dispersive X-ray (EDX) confirmed a successful loading of nHA particles on the surface of PLA@PDA after 3 and 7 days of coating (PLA@PDA-HA3 and PLA@PDA-HA7), while the surface micromorphology and porosity remain unchanged after surface modification. The thermogravimetric analysis (TGA) showed that 7.7 % and 12.3% mass ratio of nHA were loaded on the PLA scaffold surface, respectively. The wettability test indicated that the hydrophilicity of nHA-coated scaffolds was greatly enhanced, while the mechanical properties remained uncompromised. The 3D laser scanning confocal microscope (3DLS) images revealed that the surface roughness was significantly increased, reaching Sa (arithmetic mean height) of 0.402 μm in PLA@PDA-HA7. Twenty-eight days of in-vitro degradation results showed that the introduction of nHA to the PLA surface enhances its degradation properties, as evidenced by the SEM images and weight loss test. Furthermore, a sustainable release of Ca2+ from PLA@PDA-HA3 and PLA@PDA-HA7 was recorded, during the degradation process. In contrast, the released hydroxyl group of nHA tends to neutralize the local acidic environments, which was more conducive to osteoblastic differentiation and extracellular mineralization. Taken together, this facile surface modification provides 3D printed PLA scaffolds with effective bone regenerative properties by depositing Ca2+ contents, improving surface hydrophilicity, and enhancing the in-vitro degradation rate.

Study on properties of 3D-printed GelMA hydrogel scaffolds with different nHA contents

Biological 3D printing is a reliable technology for 3D printing bone repair scaffolds with simple operation, high efficiency, and relatively low cost. Gelatin methacryloyl (GelMA) hydrogels have attracted much attention due to their good biocompatibility, but the poor mechanical properties limit their application in bone reconstruction engineering. In this study, nano-hydroxyapatite (nHA) particles were added to GelMA hydrogels, and the performances of composite hydrogel scaffolds with different nHA contents were investigated in terms of rheological properties, light transmission properties, surface morphology, mechanical properties, and biocompatibility. The experimental results showed that the incorporation of nHA particles could effectively improve the printability and mechanical properties of the scaffolds, the scaffold fibers had better resistance to deformation, improved degradation rate, and biological experiments confirmed that nHA particles had no significant cytotoxicity. However, the addition of HA particles also reduced the light transmission properties of the slurry, and when its content exceeds a certain value, the hydrogel scaffolds show incomplete curing and eventually affect their test performance. The results can offer guidance and reference for the selection of ink and function for 3D printing bone repair scaffold.

Fabrication and characterization of nano-hydroxyapatite particles and assessment of the effect of their supplementation on growth of bacterial root endosymbionts of cowpea

The overall goal of this research was to evaluate the effect of nanohydroxyapatite (nHA) particles on the growth of endophytic root nodule symbionts of vegetable cowpea. The synthesized nHA exhibited spherical to hexagonal shaped morphology, the size range of 6–15 nm, and had irregular aggregate structures as confirmed by scanning and transmission electron microscopy techniques. The XRD and FTIR spectroscopy studies revealed the nano-crystallite nature and presence of phosphate functional group. Among the six nHA concentrations (0, 12, 25, 50, 75, and 100 mg L−1) the highest concentration resulted in maximum growth of Burkholderia seminalis CP-RN-4 and reference culture as observed by Visible spectroscopy and viable cell count analysis. Scanning Electron Microscopy analysis of broth cultured bacterial cells showed alteration in cell size in both cultures in presence of nHA besides production of copious amount of exopolysaccharide indicated through formation of cell aggregates/biofilm primordia.

Bi-layered PLGA electrospun membrane with occlusive and osteogenic properties for periodontal regeneration

Guided tissue regeneration (GTR) membranes not only can hamper undesirable tissues down-growth into the defects but also can selectively promote the in-growth of regenerative bone tissue, playing a critical role in periodontal regeneration. Herein, a bi-layered electrospun membrane with different sized pores was designed and fabricated by adjusting electrospinning parameters combing with facile two-step electrospinning. The small-sized pore layer (SL) as occlusive layer consisted of electrospun poly (lactic-co-glycolic acid) (PLGA) nanofibers, while the macroporous osteoconductive layer (ML) was attained via introducing the nano-hydroxyapatite (nHA) particles into PLGA nanofibers during electrospinning. Morphological results such as surface topography, nanofiber size, and pore size distribution, showed that the SL exhibited a dense structure with pore size mainly from 4 to 7 μm. In contrast, the ML possessed a loosely packed structure with pore size mainly from 20 to 28 μm, which was beneficial to the infiltration of the cells. Fourier transform infrared spectroscopy (FTIR), Energy dispersive spectrometer (EDS), and X-ray diffractometry (XRD) results showed that nHA particles were evenly loaded in PLGA nanofibers. In vitro biodegradation tests suggested that the bi-layered membrane possessed a proper degradation timeframe, which must function for at least 4 to 6 weeks. The cell experiments indicated that the bi-layered electrospun membrane possessed good cytocompatibility and proved the effective barrier potency of the small-sized pore layer. Furthermore, as revealed by the alkaline phosphate activity test, the PLGA/nHA layer possessed an improved osteogenic capacity for Human osteosarcoma cells (MG63). These results indicate that the bi-layered electrospun membrane may have potential for periodontal tissue regeneration.

Influence of UV exposure on mechanical behavior and cellular compatibility of nano-hydroxyapatite reinforced ultra-high molecular weight polyethylene

This work aims to improve mechanical behavior of microwave-assisted compression (MAC) molded nano-hydroxyapatite (nHA) reinforced ultra-high molecular weight polyethylene (UHMWPE) composite using ultraviolet (UV) C radiation exposure. The UHMWPE/nHA composite specimens were molded at 10%, 15%, and 20% weight fraction of nHA. The UV C radiation of intensity 0.025 J/cm2 was applied to the molded specimens of neat UHMWPE and UHMWPE/nHA composites. The UV C exposure resulted in increase of Young’s modulus, Vicker’s hardness, ultimate tensile strength, and flexural strength of neat UHMWPE by 46%, 84%, 6%, and 7%, respectively. Similar properties of UHMWPE/nHA composite decreased due to restricted mobility by nHA particles during crystallization effect caused by UV C radiation exposure. A 36% increase in the degree of crystallinity (DOC) was recorded after UV C radiation exposure on neat UHMWPE. At the same time, the DOC of UHMWPE/nHA composite decreased after UV C exposure. The cross-linking in UHMWPE was examined using Fourier-transform infrared spectroscopy (FTIR). The cellular compatibility of specimens was examined under human embryonic kidney cells (HEK293T) environment. It was found that UV C radiation exposure did not produce any adverse effect on the cellular compatibility of the composite specimens.

Fabrication of In Situ Grown Hydroxyapatite Nanoparticles Modified Porous Polyetheretherketone Matrix Composites to Promote Osteointegration and Enhance Bone Repair.

The repairment of critical-sized bone defects is a serious problem that stimulates the development of new biomaterials. In this study, nanohydroxyapatite (nHA)-doped porous polyetheretherketone (pPEEK) were successfully fabricated by the thermally induced phase separation method and hydrothermal treatment. Structural analysis was performed by X-ray diffraction. The water contact angles and scanning electron microscopy were measured to assess physical properties of surfaces. The mechanical strength of the composites is also determined. Microcomputed tomography is used to characterize the nHA content of the composites. The in vitro bioactivity of the composites with or without nHA was investigated by using murine pre-osteoblasts MC3T3-E1, and the results of cytotoxicity and cell proliferation assays revealed that the cytocompatibility of all specimens was good. Adherence assays were employed to examine the adhesion and morphology of cells on different materials. However, nHA-doped composites induced cell attachment and cell spreading more significantly. Osteogenic differentiation was investigated using alkaline phosphatase activity and alizarin red staining, and these in vitro results demonstrated that composites containing nHA particles enhanced osteoblast differentiation. Its effectiveness for promoting osteogenesis was also confirmed in an in vivo animal experiment using a tibial defective rat model. After 8 weeks of implantation, compared to the pure PEEK and pPEEK without nHA groups, the nHA-pPEEK group showed better osteogenic activity. The results indicate that the nHA-pPEEK composites are possibly a well-designed bone substitute for critical-sized bone defects by promoting bone regeneration and osteointegration successfully.

Bone mineral: A trojan horse for bone cancers. Efficient mitochondria targeted delivery and tumor eradication with nano hydroxyapatite containing doxorubicin

Efficient systemic pharmacological treatment of solid tumors is hampered by inadequate tumor concentration of cytostatics necessitating development of smart local drug delivery systems. To overcome this, we demonstrate that doxorubicin (DOX), a cornerstone drug used for osteosarcoma treatment, shows reversible accretion to hydroxyapatite (HA) of both nano (nHA) and micro (mHA) size. nHA particles functionalized with DOX get engulfed in the lysosome of osteosarcoma cells where the acidic microenvironment causes a disruption of the binding between DOX and HA. The released DOX then accumulates in the mitochondria causing cell starvation, reduced migration and apoptosis. The HA+DOX delivery system was also tested in-vivo on osteosarcoma bearing mice. Locally delivered DOX via the HA particles had a stronger tumor eradication effect compared to the controls as seen by PET-CT and immunohistochemical staining of proliferation and apoptosis markers. These results indicate that in addition to systemic chemotherapy, an adjuvant nHA could be used as a carrier for intracellular delivery of DOX for prevention of tumor recurrence after surgical resection in an osteosarcoma. Furthermore, we demonstrate that nHA particles are pivotal in this approach but a combination of nHA with mHA could increase the safety associated with particulate nanomaterials while maintaining similar therapeutic potential.