Incorporation Of nHA Research Articles

Fostering biomineralization and biodegradation: nano-hydroxyapatite reinforced iron composites for biodegradable implant application

Iron (Fe) is regarded as a candidate material for biodegradable metallic implants due to its biocompatibility and ability to degrade in physiological environemnt. However, the degradation rate in the physiological environment is too slow for clinical applications. It is necessary to accelerate the rate such that the degradation is compatible with tissue growth. Furthermore, the implant material needs to be bioactive for promoting osteointegration, osteoconductivity, cell proliferation and biological apatite formation. Nano-sized bioactive hydroxyapatite (nHA) was incorporated into a porous Fe matrix to enhance the bioactivity and degradation rate. Electrochemical studies in biomemtic NaCl solution, revealed that incorporation of nHA in porous Fe can increase the degradation by 2.5 times compared to the pure iron counterparts with similar porosity. Furthermore, immersion tests in simulated body fluid (SBF) revealed that nHA added samples displayed enhanced biomineralization and degraded at a rate three times faster than pure iron in this environment. The incorporation of nHA into the iron matrix aided in the formation of biomineralized hydroxyapatite. The composite surface promoted the cell adhesion and proliferation and the L929 fibroblast cells exhibited good cell viability. It is proposed that porous morphology incorporated with nHA can improve biomineralization and tailor the degradation rate of Fe-based materials in physiological environment.

Dual-functional thermosensitive hydrogel for reducing infection and enhancing bone regeneration in infected bone defects

The contamination of bone defects is a serious therapeutic problem. The treatment of infected bone defects involves rigorous infection control followed by bone reconstruction. Considering these two processes, the development of biomaterials possessing antibacterial and osteogenic properties offers a promising approach for the treatment of infected bone defects. In this study, a dual-functional, thermosensitive, and injectable hydrogel composed of chitosan (CS), quaternized CS (QCS), and nano-hydroxyapatite (nHA) was designed, and the ratio of CS to QCS in the hydrogel was optimized to enhance the antibacterial efficacy of CS while reducing the cytotoxicity of QCS. In vitro studies demonstrated that the hydrogel with an 85 %:15 % ratio of CS to QCS exhibited excellent biocompatibility and antibacterial properties while also possessing suitable mechanical characteristics and degradability. The incorporation of nHA into the hydrogel enhanced MC3T3-E1 proliferation and osteogenic differentiation. Moreover, this hydrogel demonstrated superior in vivo therapeutic effectiveness in a rabbit model of infected bone defect. In summary, this study provides a promising material design and a comprehensive one-step treatment strategy for infected bone defects.

A bilayer hydrogel mimicking the periosteum-bone structure for innervated bone regeneration.

In bone tissue, nerves are primarily located in the periosteum and play an indispensable role in bone defect repair. However, most bone tissue engineering approaches ignored the reconstruction of the nerve network. Herein, we aimed to develop a bilayer hydrogel simulating periosteum-bone structure to induce innervated bone regeneration. The bottom "bone" layer consisted of gelatin methacryloyl (GelMA), poly(ethylene glycol) diacrylate (PEGDA), and nano-hydroxyapatite (nHA), whereas the upper "periosteum" layer consisted of GelMA, sodium alginate (SA) and MgCl2. The mechanical properties of the upper and bottom hydrogels were designed to be suitable for neurogenesis and osteogenesis, respectively. Besides, Mg2+ from the "periosteum" layer released at the early stage (within 7 d), which aligned with the optimal time window for nerve regeneration and osteogenic related neuropeptide release. Simultaneously, the prevention of long-term Mg2+ release (after 7 d) could avoid osteogenic inhibition caused by prolonged Mg2+ exposure. Additionally, the incorporation of nHA in the bottom "bone" layer supported the long-term osteogenesis due to its osteoconductivity and slow degradation. In vitro biological experiments revealed that the bilayer hydrogel (GS@Mg/GP@nHA) promoted neurite growth and calcitonin gene-related peptide (CGRP) expression in rat dorsal root ganglion (DRG) neurons, as well as the osteogenesis of rat bone-derived mesenchymal stem cells (BMSCs). Moreover, the in vivo experiments demonstrated that the GS@Mg/GP@nHA hydrogel efficiently promoted nerve network reconstruction and bone regeneration of rat calvarial bone defects. Altogether, the bilayer hydrogel GS@Mg/GP@nHA could promote innervated bone regeneration, providing new insights for biomaterial design for bone tissue engineering.

Evaluation of Physicochemical Properties of a Hydroxyapatite Polymer Nanocomposite for Use in Fused Filament Fabrication.

Over the last decade, there has been an increasing interest in the use of bioceramics for biomedical purposes. Bioceramics, specifically those made of calcium phosphate, are commonly used in dental and orthopaedic applications. In this context, hydroxyapatite (HA) is considered a viable option for hard tissue engineering applications given its compositional similarity to bioapatite. However, owing to their poor mechanobiology and biodegradability, traditional HA-based composites have limited utilisation possibilities in bone, cartilage and dental applications. Therefore, the efficiency of nano HA (nHA) has been explored to address these limitations. nHA has shown excellent remineralising effects on initial enamel lesions and is widely used as an additive for improving existing dental materials. Furthermore, three-dimensional printing (3DP) or fused deposition modelling that can be used for creating dental and hard tissue scaffolds tailored to each patient's specific anatomy has attracted considerable interest. However, the materials used for producing hard tissue with 3DP are still limited. Therefore, the current study aimed to develop a hybrid polymer nanocomposite composed of nHA, nanoclay (NC) and polylactic acid (PLA) that was suitable for 3DP. The nHA polymer nanocomposites were extruded into filaments and their physiochemical properties were evaluated. The results showed that the addition of nHA and NC to the PLA matrix significantly increased the water absorption and contact angle. In addition, the hardness increased from 1.04 to 1.25 times with the incorporation of nHA. In sum, the nHA-NC-reinforced PLA could be used as 3DP filaments to generate bone and dental scaffolds, and further studies are needed on the biocompatibility of this material.

Characterization of PLA/PCL/Nano-Hydroxyapatite (nHA) Biocomposites Prepared via Cold Isostatic Pressing.

Hydroxyapatite has the closest chemical composition to human bone. Despite this, the use of nano-hydroxyapatite (nHA) to produce biocomposite scaffolds from a mixture of polylactic acid (PLA) and polycaprolactone (PCL) using cold isostatic pressing has not been studied intensively. In this study, biocomposites were created employing nHA as an osteoconductive filler and a polymeric blend of PLA and PCL as a polymer matrix for prospective usage in the medical field. Cold isostatic pressing and subsequent sintering were used to create composites with different nHA concentrations that ranged from 0 to 30 weight percent. Using physical and mechanical characterization techniques such as Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM), and density, porosity, tensile, and flexural standard tests, it was determined how the nHA concentrations affected the biocomposite's general properties. In this study, the presence of PLA, PCL, and nHA was well identified using FTIR, XRD, and SEM methods. The biocomposites with high nHA content showed intense bands for symmetric stretching and the asymmetric bending vibration of PO43-. The incorporation of nHA into the polymeric blend matrix resulted in a rather irregular structure and the crystallization became more difficult. The addition of nHA improved the density and tensile and flexural strength of the PLA/PCL matrix (0% nHA). However, with increasing nHA content, the PLA/PCL/nHA biocomposites became more porous. In addition, the density, flexural strength, and tensile strength of the PLA/PCL/nHA biocomposites decreased with increasing nHA concentration. The PLA/PCL/nHA biocomposites with 10% nHA had the highest mechanical properties with a density of 1.39 g/cm3, a porosity of 1.93%, a flexural strength of 55.35 MPa, and a tensile strength of 30.68 MPa.

P8 - Polycaprolactone/Nano-Hydroxyapatite Nanofibrous Scaffold as a Cell Homing-Based Therapy

This study investigated the behavior of human dental pulp cells (HDPCs) seeded on the surface of polycaprolactone/nano-hydroxyapatite nanofibrous scaffolds, aiming a dentin tissue engineering-based vital pulp therapy (VPT). Polycaprolactone (PCL)-based solutions (10% w/v in 8:2 chloroform and dimethylformamide) containing nano-hydroxyapatite (nHA) particles (0.5; 1.0; or 2.0% w/v) were electrospun into nanofibrous scaffolds. PCL scaffolds without nHA were used as control. The scaffolds were blindly characterized for morphology and composition (SEM/EDS; n=12), solubility (n=6), the release of calcium/phosphate (n=6), and change of medium pH (n=6). A culture of HDPCs was obtained from sound third molars and characterized for the presence of a subpopulation of mesenchymal stem cells. Then, HDPCs were seeded on the surface of the scaffolds and evaluated for cell viability (alamarBlue; n=8), cell proliferation (Live/Dead assay; n=4), adhesion and spreading (F-actin; n=4), total protein content (TP; Lowry method; n=8), alkaline phosphatase activity (ALP; thymolphthalein assay; n=8), expression of odontogenic genes (RT-qPCR; DSPP, DMP1, COL1A1, and ALPL; n=6), and formation of a mineralized matrix (Alizarin Red staining; n=8). All experiments were repeated at least twice. Sample sizes were calculated using G-Power. Data were analyzed with confidence intervals and one- or two-way ANOVA followed by Tukey, Sidak, or Games-Howell post-hocs (?=5%). All formulations generated randomly arranged fibers ranging from 600 to 900 nm in diameter. Higher nHA concentrations roughened the surface of the nanofibers, while PCL+2%nHA increased the interfibrillar spaces. PCL+1%nHA and PCL+2%nHA significantly released calcium/phosphate over a period of 28 days in comparison with PCL+0.5%nHA. Nonetheless, the medium pH was maintained below 8.0. HDPCs viability and proliferation were not affected by the incorporation of nHA, while cell adhesion/spreading was stimulated, especially for PCL+2%nHA. Higher total protein content and increased ALP activity were seen in the presence of nHA. PCL+1%nHA and PCL+2%nHA upregulated the expression of DSPP and DMP1 in 14 days; and COL1A1, ALPL, and DMP1 in 21 days. The formation of a mineralized matrix after 21 days was concentration-dependent, and it was about 9× higher for PCL+2%nHA formulation compared to control.

Nanohydroxyapatite embedded blends of ethylene‐co‐vinyl acetate and millable polyurethane as piezoelectric materials: dielectric, viscoelastic and mechanical features

AbstractWe report the piezoelectric, dielectric and dynamic mechanical features of nanohydroxyapatite (nHA) embedded ethylene‐co‐vinyl acetate (EVA)/millable polyurethane (MPU) elastomer blends. The EVA/MPU/nHA systems were prepared by two‐roll milling followed by compression moulding. The piezoelectric features were probed by dynamic contact electrostatic force microscopy (EFM).The neat EVA/MPU blend was not found to exhibit ferroelectric properties. However, upon incorporation of nHA, butterfly loops of EFM amplitude signals as well as complementary hysteresis curves were obtained, confirming the introduction of ferro‐piezoelectric properties. The ferro‐piezoelectric features were found to be improved with an increase in nHA loading which recorded highest signals for 8 parts per hundred parts of rubber (phr) loaded composites. The fabricated devices generated a peak voltage of 4.02 V and 4.89 V respectively with 6 phr and 8 phr nHA loaded composites. Electrical properties such as dielectric constant, volume resistivity and loss factor as well as dynamic mechanical properties were examined. The viscoelastic features of the systems support the reinforcing feature with filler loading. TEM analysis of the composites indicated a uniform distribution of filler particles in the blend matrix, complementing the observed ferro‐piezoelectric, dielectric and dynamic mechanical properties. The simultaneous modulation of piezoelectric and viscoelastic properties is a preferred feature for fast‐responsive dielectric elastomers for their applications in various piezoelectric devices. © 2020 Society of Chemical Industry

Synthetic Biodegradable Aliphatic Polyester Nanocomposites Reinforced with Nanohydroxyapatite and/or Graphene Oxide for Bone Tissue Engineering Applications.

This paper provides review updates on the current development of bionanocomposites with polymeric matrices consisting of synthetic biodegradable aliphatic polyesters reinforced with nanohydroxyaptite (nHA) and/or graphene oxide (GO) nanofillers for bone tissue engineering applications. Biodegradable aliphatic polyesters include poly(lactic acid) (PLA), polycaprolactone (PCL) and copolymers of PLA-PGA (PLGA). Those bionanocomposites have been explored for making 3D porous scaffolds for the repair of bone defects since nHA and GO enhance their bioactivity and biocompatibility by promoting biomineralization, bone cell adhesion, proliferation and differentiation, thus facilitating new bone tissue formation upon implantation. The incorporation of nHA or GO into aliphatic polyester scaffolds also improves their mechanical strength greatly, especially hybrid GO/nHA nanofilllers. Those mechanically strong nanocomposite scaffolds can support and promote cell attachment for tissue growth. Porous scaffolds fabricated from conventional porogen leaching, and thermally induced phase separation have many drawbacks inducing the use of organic solvents, poor control of pore shape and pore interconnectivity, while electrospinning mats exhibit small pores that limit cell infiltration and tissue ingrowth. Recent advancement of 3D additive manufacturing allows the production of aliphatic polyester nanocomposite scaffolds with precisely controlled pore geometries and large pores for the cell attachment, growth, and differentiation in vitro, and the new bone formation in vivo.

Characterization, drug loading and antibacterial activity of nanohydroxyapatite/polycaprolactone (nHA/PCL) electrospun membrane

Considering the important factor of bioactive nanohydoxyapatite (nHA) to enhance osteoconductivity or bone-bonding capacity, nHA was incorporated into an electrospun polycaprolactone (PCL) membrane using electrospinning techniques. The viscosity of the PCL and nHA/PCL with different concentrations of nHA was measured and the morphology of the electrospun membranes was compared using a field emission scanning electron microscopy. The water contact angle of the nanofiber determined the wettability of the membranes of different concentrations. The surface roughness of the electrospun nanofibers fabricated from pure PCL and nHA/PCL was determined and compared using atomic force microscopy. Attenuated total reflectance Fourier transform infrared spectroscopy was used to study the chemical bonding of the composite electrospun nanofibers. Beadless nanofibers were achieved after the incorporation of nHA with a diameter of 200–700 nm. Results showed that the fiber diameter and the surface roughness of electrospun nanofibers were significantly increased after the incorporation of nHA. In contrast, the water contact angle (132° ± 3.5°) was reduced for PCL membrane after addition of 10% (w/w) nHA (112° ± 3.0°). Ultimate tensile strengths of PCL membrane and 10% (w/w) nHA/PCL membrane were 25.02 ± 2.3 and 18.5 ± 4.4 MPa. A model drug tetracycline hydrochloride was successfully loaded in the membrane and the membrane demonstrated good antibacterial effects against the growth of bacteria by showing inhibition zone for E. coli (2.53 ± 0.06 cm) and B. cereus (2.87 ± 0.06 cm).

Assembling of electrospun meshes into three-dimensional porous scaffolds for bone repair

Technical limitations of traditional electrospinning make it hard to produce three-dimensional (3D) scaffolds with hierarchical pore structures. Here, porous polycaprolactone (PCL) nanofiber meshes with different nano-hydroxyapatite (nHA) concentrations were prepared by electrospinning with stainless steel mesh as the collector, and 3D porous nanofiber scaffolds were fabricated via layer-by-layer assembly with a special binder (18% PCL/DCM solution). The single layer nanofiber mesh possessed very regular morphology with a hollow structure, and the nHA was not only embedded in the nanofiber but also exposed on the surfaces of the fiber, resulting in the improved surface chemical properties. The incorporation of nHA also had a significant effect on cell behaviours and functions. The 3D nanofiber scaffolds possessed hierarchical structures with interconnected micro and macro pores, which allowed cells to migrate between the adjacent layers, even throughout the scaffold. Cells filled the scaffold space and integrated with the nanofiber materials, forming scaffold/cells complexes in vitro. In addition, alendronate was successfully carried on the 3D composite scaffolds because of the high affinity of P–C–P backbone to calcium ions. The composite scaffolds treated with alendronate significantly promote the osteogenesis-related gene expression of human foetal osteoblasts. All these results suggest that 3D functional nanofiber scaffolds would be potentially useful for bone repair.

PEDOT:PSS-Containing Nanohydroxyapatite/Chitosan Conductive Bionanocomposite Scaffold: Fabrication and Evaluation

Conductive poly(3,4-ethylenedioxythiophene)-poly(4-styrene sulfonate) (PEDOT:PSS) was incorporated into nanohydroxyapatite/chitosan (nHA/CS) composite scaffolds through a freezing and lyophilization technique. The bionanocomposite conductive scaffold was then characterized using several techniques. A scanning electron microscope image showed that the nHA and PEDOT:PSS were dispersed homogeneously in the chitosan matrix, which was also confirmed by energy-dispersive X-ray (EDX) analysis. The conductive properties were measured using a digital multimeter. The weight loss and water-uptake properties of the bionanocomposite scaffolds were studiedin vitro. Anin vitrocell cytotoxicity test was carried out using mouse fibroblast (L929) cells cultured onto the scaffolds. Using a freezing and lyophilization technique, it was possible to fabricate three-dimensional, highly porous, and interconnected PEDOT:PSS/nHA/CS scaffolds with good handling properties. The porosity was 74% and the scaffold’s conductivity was9.72±0.78 μS. The surface roughness was increased with the incorporation of nHA and PEDOT:PSS into the CS scaffold. The compressive mechanical properties increased significantly with the incorporation of nHA but did not change significantly with the incorporation of PEDOT:PSS. The PEDOT:PSS-containing nHA/CS scaffold exhibited significantly higher cell attachment. The PEDOT:PSS/nHA/CS scaffold could be a potential bionanocomposite conductive scaffold for tissue engineering.

The hydroxyapatite nanotube as a promoter to optimize the HDS reaction of NiMo/TiO2 catalyst

A composite containing TiO2 whiskers (TiO2(w)) and hydroxyapatite nanotube (nHA) with mosaic structure functioned as a support was successfully synthesized by a facile route. The TiO2(w) was prepared from potassium titanate and the novel single-crystalline hydroxyapatite nanotubes were synthesized via a distinctive biomimetic route. Properties of support and catalyst were characterized by SEM, TEM, XRD, FT-IR of adsorbed pyridine (in situ FT-IR), BET, TPR, and NH3-TPD. Catalytic activity of NiMo/TiO2(w)-nHA, and the effect of nHA contents on hydrodesulfurization (HDS) activity was performed. Results indicated the incorporation of nHA can weaken the Lewis acidity of TiO2 supports and decrease acidity of the NiMo/TiO2(w)-nHA catalyst. The addition of moderate nHA has weak influence on the textural and structural properties of the catalyst. NiMo/TiO2(w)-nHA exhibits higher HDS catalytic activity than that of NiMo/TiO2(w) catalyst under mild conditions of low temperature and pressure (280°C and 2MPa), and the catalytic performance of NiMo/TiO2(w)-nHA-0.1 is the best. The synergistic effects between TiO2(w) and nHA and acidity regulation contribute to improve the HDS catalytic activity of the nHA-doped samples.

Preparation, characterization, and in vitro osteoblast functions of a nano-hydroxyapatite/polyetheretherketone biocomposite as orthopedic implant material.

A bioactive composite was prepared by incorporating 40 wt% nano-hydroxyapatite (nHA) into polyetheretherketone (PEEK) through a process of compounding, injection, and molding. The mechanical and surface properties of the nHA/PEEK composite were characterized, and the in vitro osteoblast functions in the composite were investigated. The mechanical properties (elastic modulus and compressive strength) of the nHA/PEEK composite increased significantly, while the tensile strength decreased slightly as compared with PEEK. Further, the addition of nHA into PEEK increased the surface roughness and hydrophilicity of the nHA/PEEK composite. In cell tests, compared with PEEK and ultra-high-molecular-weight polyethylene, it was found that the nHA/PEEK composite could promote the functions of MC3T3-E1 cells, including cell attachment, spreading, proliferation, alkaline phosphatase activity, calcium nodule formation, and expression of osteogenic differentiation-related genes. Incorporation of nHA into PEEK greatly improved the bioperformance of PEEK. The nHA/PEEK composite might be a promising orthopedic implant material.

Fabrication and in vitro biocompatibility of biomorphic PLGA/nHA composite scaffolds for bone tissue engineering

In this study, biomorphic poly(dl-lactic-co-glycolic acid)/nano-hydroxyapatite (PLGA/nHA) composite scaffolds were successfully prepared using cane as a template. The porous morphology, phase, compression characteristics and in vitro biocompatibility of the PLGA/nHA composite scaffolds and biomorphic PLGA scaffolds as control were investigated. The results showed that the biomorphic scaffolds preserved the original honeycomb-like architecture of cane and exhibited a bimodal porous structure. The average channel diameter and micropore size of the PLGA/nHA composite scaffolds were 164±52μm and 13±8μm, respectively, with a porosity of 89.3±1.4%. The incorporation of nHA into PLGA decreased the degree of crystallinity of PLGA, and significantly improved the compressive modulus of biomorphic scaffolds. The in vitro biocompatibility evaluation with MC3T3-E1 cells demonstrated that the biomorphic PLGA/nHA composite scaffolds could better support cell attachment, proliferation and differentiation than the biomorphic PLGA scaffolds. The localization depth of MC3T3-E1 cells within the channels of the biomorphic PLGA/nHA composite scaffolds could reach approximately 400μm. The results suggested that the biomorphic PLGA/nHA composite scaffolds were promising candidates for bone tissue engineering.

The performance of dental pulp stem cells on nanofibrous PCL/gelatin/nHA scaffolds

The aim of current study is to investigate the in vitro and in vivo behavior of dental pulp stem cells (DPSCs) seeded on electrospun poly(epsilon-caprolactone) (PCL)/gelatin scaffolds with or without the addition of nano-hydroxyapatite (nHA). For the in vitro evaluation, DNA content, alkaline phosphatase (ALP) activity and osteocalcin (OC) measurement showed that the scaffolds supported DPSC adhesion, proliferation, and odontoblastic differentiation. Moreover, the presence of nHA upregulated ALP activity and promoted OC expression. Real-time PCR data confirmed these results. SEM micrographs qualitatively confirmed the proliferation and mineralization characteristics of DPSCs on both scaffolds. Subsequently, both scaffolds seeded with DPSCs were subcutaneously implanted into immunocompromised nude mice. Scaffolds with nHA but without cells were implanted as control. Histological evaluation revealed that all implants were surrounded by a thin fibrous tissue capsule without any adverse effects. The cell/scaffold composites showed obvious in vivo hard tissue formation, but there was no sign of tissue ingrowth. Further, the combination of nHA in scaffolds did upregulate the expression of specific odontogenic genes. In conclusion, the incorporation of nHA in nanofibers indeed enhanced DPSCs differentiation towards an odontoblast-like phenotype in vitro and in vivo.

Preparation and characterization of nano-hydroxyapatite/polyamide 66 composite GBR membrane with asymmetric porous structure

In this study, a nano-hydroxyapatite/polyamide 66 (nHA/PA66) composite with good biocompatibility and high bioactivity is employed to develop novel asymmetric structure porous membranes for guided bone regeneration (GBR). FT-IR and XRD analyses suggest that chemical bonds are formed between nHA and PA66 both in composite powders and membranes. The fabricated membranes show gradient porous structure. SEM analysis reveal that pores less than 10 microm and pores with a size ranging from 30 microm to 200 microm distribute in the micropore layer and the spongy structure layer, respectively. The surface energy determination also reveals that the fabricated membranes have asymmetric surface properties on the two sides of the membrane. The incorporation of nHA in PA66 matrix improves the properties of the membrane. The elongation at break and the tensile strength of nHA/PA66-40 suggest that the composite membrane has good strength and toughness. The rough porous structure surface with high surface energy of nHA/PA66 composite membrane may be beneficial to promote cells immobility and differentiation into a mature phenotype producing mineralized matrix. The biocompatibility, bioactivity, osteoconductivity, asymmetric porous structure, mechanical properties and hydrophilicity of the composite membrane can meet the requirement of GBR technique.