Evaluation of physico-mechanical properties and in vitro biocompatibility of compression molded HDPE based biocomposites with HA/Al2O3 ceramic fillers and titanate coupling agents

The present article demonstrates how the stiffness, hardness as well as the cellular response of bioinert high-density polyethylene (HDPE) can be significantly improved with combined addition of both bioinert and bioactive ceramic fillers. For this purpose, different amounts of hydroxyapatite and alumina, limited to a total of 40 wt %, have been incorporated in HDPE matrix. An important step in composite fabrication was to select appropriate solvent and optimal addition of coupling agent (CA). In case of chemically coupled composites, 2% Titanium IV, 2-propanolato, tris iso-octadecanoato-O was used as a CA. All the hybrid composites, except monolithic HDPE, were fabricated under optimized compression molding condition (140 degrees C, 0.75 h, 10 MPa pressure). The compression molded composites were characterized, using X-ray diffraction, Fourier transformed infrared spectroscopy, and scanning electron microscopy. Importantly, in vitro cell culture and cell viability study (MTT) using L929 fibroblast and SaOS2 osteoblast-like cells confirmed good cytocompatibility properties of the developed hybrid composites. (C) 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2012

Dependence of Protein Adsorption on Wetting Behavior of UHMWPE-HA-Al2O3-CNT Hybrid Biocomposites

Ultrahigh-molecular-weight polyethylene (UHMWPE) is used as an articulating surface in total hip and knee joint replacement. In order to enhance long-term durability/wear resistance properties, UHMWPE-based polymer-ceramic hybrid composites are being developed. Surface properties such as wettability and protein adsorption alter with reinforcement or with change in surface chemistry. From this perspective, the wettability and protein adsorption behavior of compression-molded UHMWPE-hydroxyapatite (HA)-aluminum oxide (Al2O3)-carbon nanotube (CNT) composites were analyzed in conjunction with surface roughness. The combined effect of Al2O3 and CNT shows enhancement of the contact angle by similar to 37A degrees compared with the surface of the UHMWPE matrix reinforced with HA. In reference to unreinforced UHMWPE, protein adsorption density also increased by similar to 230% for 2 wt.%HA-5 wt.%Al2O3-2 wt.%CNT addition to UHMWPE. An important conclusion is that the polar and dispersion components of the surface free energy play a significant role in wetting and protein adsorption than do the total free energy or chemistry of the surface. The results of this study have major implications for the biocompatibility of these newly developed biocomposites.

In vitro cytotoxicity and in vivo osseointergration properties of compression-molded HDPE-HA-Al2O3 hybrid biocomposites

The aim of this study was to investigate the in vivo biocompatibility in terms of healing of long segmental bone defect in rabbit model as well as in vitro cytotoxicity of eluates of compression-molded High density polyethylene (HDPE)hydroxyapatite (HA)-aluminum oxide (Al2O3) composite-based implant material. Based on the physical property in terms of modulus and strength properties, as reported in our recent publication, HDPE-40 wt % HA and HDPE-20 wt % HA-20 wt % Al2O3 hybrid composites were used for biocompatibility assessment. Osteoblasts cells were cultured in conditioned media, which contains varying amount of composite eluate (0.01, 0.1, and 1.0 wt %). In vitro, the eluates did not exhibit any significant negative impact on proliferation, mineralization or on morphology of human osteoblast cells. In vivo, the histological assessment revealed neobone formation at the bone/implant interface, characterized by the presence of osteoid and osteoblasts. The observation of osteoclastic activity indicates the process of bone remodeling. No inflammation to any noticeable extent was observed at the implantation site. Overall, the combination of in vitro and in vivo results are suggestive of potential biomedical application of compression-molded HDPE- 20 wt % HA- 20 wt % Al2O3 composites to heal long segmental bone defects without causing any toxicity of bone cells.

Flow cytometry analysis of human fetal osteoblast fate processes on spark plasma sintered hydroxyapatite-titanium biocomposites

Hydroxyapatite (HA)-based biocomposites have been widely investigated for a multitude of applications and these studies have been largely driven to improve mechanical properties (toughness and strength) without compromising cytocompatibility properties. Apart from routine cell viability/proliferation analysis, limited efforts have been made to quantify the fate processes (cell proliferation, cell cycle, and cell apoptosis) of human fetal osteoblast (hFOB) cells on HA-based composites, in vitro. In this work, the osteoblast cell fate process has been studied on a model hydroxyapatite-titanium (HA-Ti) system using the flow cytometry. In order to retain both HA and Ti, the novel processing technique, that is, spark plasma sintering, was suitably adopted. The cell fate processes of hFOBs, as evaluated using a flow cytometry, revealed statistically insignificant differences among HA-10 wt % Ti and HA and control (tissue culture polystyrene surface) in terms of osteoblast apoptosis, proliferation index as well as division index. For the first time, we provide quantified flow cytometry results to demonstrate that 10 wt % Ti additions to HA do not have any significant influence on the fate processes of human osteoblast-like cells, in vitro.

Spark Plasma Sintered HA-Fe3O4-Based Multifunctional Magnetic Biocomposites

Although HA is highly biocompatible, one of the major disadvantages of HA include the lack of antibacterial property. In an earlier study, we demonstrated the potential role of magnetic field stimulation on bactericidal property in vitro. Following this, it was hypothesized that antibacterial property can be realized if bacteria are grown on magnetic biocomposites in vitro. In addressing this issue, this study demonstrates the development of HA-Fe3O4-based magnetic substrate with multifunctional properties. For this purpose, HA-xFe(3)O(4) (x: 10, 20 and 40wt%) powder compositions were sintered using uniquely designed spark plasma sintering conditions (three stage sintering with final holding temperature of 1050 degrees C for 5min). A saturation magnetization of 24emu/g is measured with HA-40%Fe3O4. Importantly, all the HA-Fe3O4 composites demonstrated bactericidal property by rupturing the membrane of Escherichia coli bacteria, while supporting cell growth of metabolically active human fetal osteoblast cells over 8d culture. A systematic decrease in bacterial viability with Fe3O4 addition is consistent with a commensurate increase in reactive oxygen species (ROS).

Mechanical and Thermal Properties of Eva Blended with Biodegradable Ethyl Cellulose

In this study, biodegradable blend of Poly (Ethylene-co-Vinyl Acetate) (EVA) and Ethyl Cellulose (EC) were prepared. Ethylene vinyl alcohol (EVOH) copolymer was used as an interfacial compatibilizer to enhance adhesion between EVA and EC. The melt blended compatibilized biocomposites were examined for mechanical and thermal properties as per the ASTM standards. It has been found that the EC has a reinforcing effect on EVA leading to enhanced tensile strength and also impart biodegradability. Thus, a high loading of 50% EC could be added without compromising Much on the mechanical properties. Analysis of the tensile data using predictive theories showed an enhanced interaction of the dispersed phase (EC) and the matrix (EVA). The compatibilizing effects of EVOH on these blends were confirmed by the significant improvement in the mechanical properties comparable with neat EVA as also observed by SEM microscopy. The TGA thermograms exhibits two-stage degradation and as EC content increases, the onset temperature for thermal degradation reduces. (C) 2009 Wiley Periodicals, Inc. J Appl Polym Sci 116: 1044-1056, 2010

Injection-molded high-density polyethylene-hydroxyapatite-aluminum oxide hybrid composites for hard-tissue replacement: Mechanical, biological, and protein adsorption behavior

In this article, we report the mechanical and biocompatibility properties of injection-molded high-density polyethylene (HDPE) composites reinforced with 40 wt % ceramic filler [hydroxyapatite (HA) and/or Al2O3] and 2 wt % titanate as a coupling agent. The mechanical property measurements revealed that a combination of a maximum tensile strength of 18.7 MPa and a maximum tensile modulus of about 855 MPa could be achieved with the injection-molded HDPE20 wt % HA20 wt % Al2O3 composites. For the same composite composition, the maximum compression strength was determined to be 71.6 MPa and the compression modulus was about 660 MPa. The fractrography study revealed the uniform distribution of ceramic fillers in the semicrystalline HDPE matrix. The cytocompatibility study with osteoblast-like SaOS2 cells confirmed extensive cell adhesion and proliferation on the injection-molded HDPE20 wt % HA20 wt % Al2O3 composites. The cell viability analysis with the 3(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay revealed a statistically significant difference between the injection-molded HDPE20 wt % HA20 wt % Al2O3 composites and sintered HA for various culture durations of upto 7 days. The difference in cytocompatibility properties among the biocomposites is explained in terms of the difference in the protein absorption behavior. (C) 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2012

Substrate conductivity dependent modulation of cell proliferation and differentiation in vitro

In designing and developing various biomaterials, the influence of substrate properties, like surface topography, stiffness and wettability on the cell functionality has been investigated widely. However, such study to probe into the influence of the substrate conductivity on cell fate processes is rather limited. In order to address this issue, spark plasma sintered HA-CaTiO3 (Hydroxyapatite-Calcium titanate) has been used as a model material system to showcase the effect of varying conductivity on cell functionality. Being electroactive in nature, mouse myoblast cells (C2C12) were selected as a model cell line in this study. It was inferred that myoblast adhesion/growth systematically increases with substrate conductivity due to CaTiO3 addition to HA. Importantly, parallel arrangement of myoblast cells on higher CaTiO3 containing substrates indicate that self-adjustable cell patterning can be achieved on conductive biomaterials. Furthermore, enhanced myoblast assembly and myotube formation were recorded after 5 days of serum starvation. Overall, the present study conclusively establishes the positive impact of the substrate conductivity towards cell proliferation and differentiation as well as confirms the efficacy of HA-CaTiO3 biocomposites as conductive platforms to facilitate the growth, orientation and fusion of myoblasts, even when cultured in the absence of external electric field.

On the toughness enhancement in hydroxyapatite-based composites

Among various biologically compatible materials, hydroxyapatite (HA) has excellent bioactivity/osteointegration properties and therefore has been extensively investigated for biomedical applications. However, its inferior fracture toughness limits the wider applications of monolithic HA as a load-bearing implant. To this end, HA-based biocomposites have been developed to improve their mechanical properties (toughness and strength) without compromising biocompatibility. Despite significant efforts over last few decades, the toughness of HA-based composites could not be enhanced beyond 1.5-2 MPa m(1/2), even when measured using indentation techniques. In this perspective, the present work demonstrates how spark plasma sintering can be effectively utilized to develop hydroxyapatite titanium (HA-Ti) composites with varying amounts of Ti (5, 10 and 20 wt.%) with extremely high single edge V-notch beam fracture toughness (4-5 MPa m(1/2)) along with a good combination of elastic modulus and flexural strength. Despite predominant retention of HA and Ti, the combination of critical analysis of X-ray diffraction and transmission electron microscopy investigation confirmed the formation of the CaTi4(PO4)(6) phase with nanoscale morphology at the HA/Ti interface and the formation of such a phase has been discussed in reference to possible sintering reactions. The variations in the measured fracture toughness and work of fracture with Ti addition to the HA matrix were further rationalized using the analytical models of crack bridging as well as on the basis of the additional contribution from crack deflection. The present work opens up the opportunity to further enhance the toughness beyond 5 MPa m(1/2) by microstructural designing with the desired combination of toughening phases. (C) 2013 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Fretting wear behaviour of hydroxyapatite-titanium composites in simulated body fluid, supplemented with 5 g l(-1) bovine serum albumin

Damaged articulating joints can be repaired or replaced with synthetic biomaterials, which can release wear debris due to articulation, leading to the osteolysis. In a recent work, it has been shown that it is possible to achieve a better combination of flexural strength/fracture toughness as well as in vitro bioactivity and cytocompatibility properties in spark plasma sintered hydroxyapatite-titanium (HA-Ti) composites. Although hydroxyapatite and titanium are well documented for their good biocompatibility, nanosized hydroxyapatite (HA) and titanium (Ti) particles can cause severe toxicity to cells. In order to address this issue, fretting wear study of HA-Ti composites under dry and wet (1x SBF, supplemented with 5 g l(-1) bovine serum albumin (BSA)) condition was performed to assess the wear resistance as well as wear debris formation, in vitro. The experimental results reveal one order of magnitude lower wear rate for HA-10 wt% Ti (7.5 x 10(-5) mm(3) N-1 m(-1)) composite than monolithic HA (3.9 x 10(-4) mm(3) N-1 m(-1)) in simulated body fluid. The difference in the tribological properties has been analyzed in the light of phase assemblages and mechanical properties. Overall, the results suggest the potential use of HA-Ti composites over existing HA-based biocomposites in orthopedic as well as dental applications.

Spark plasma sintered HA-Fe3O4-based multifunctional magnetic Bbiocomposites

Although HA is highly biocompatible, one of the major disadvantages of HA include the lack of antibacterial property. In an earlier study, we demonstrated the potential role of magnetic field stimulation on bactericidal property in vitro. Following this, it was hypothesized that antibacterial property can be realized if bacteria are grown on magnetic biocomposites in vitro. In addressing this issue, this study demonstrates the development of HA-Fe3O4-based magnetic substrate with multifunctional properties. For this purpose, HA-xFe(3)O(4) (x: 10, 20 and 40wt%) powder compositions were sintered using uniquely designed spark plasma sintering conditions (three stage sintering with final holding temperature of 1050 degrees C for 5min). A saturation magnetization of 24emu/g is measured with HA-40%Fe3O4. Importantly, all the HA-Fe3O4 composites demonstrated bactericidal property by rupturing the membrane of Escherichia coli bacteria, while supporting cell growth of metabolically active human fetal osteoblast cells over 8d culture. A systematic decrease in bacterial viability with Fe3O4 addition is consistent with a commensurate increase in reactive oxygen species (ROS).

Bactericidal effect of silver-reinforced carbon nanotube and hydroxyapatite composites

Bacterial infection remains an important risk factor after orthopedic surgery. The present paper reports the synthesis of hydroxyapatite-silver (HA-Ag) and carbon nanotube-silver (CNT-Ag) composites via spark plasma sintering (SPS) route. The retention of the initial phases after SPS was confirmed by phase analysis using X-ray diffraction and Raman spectroscopy. Energy dispersive spectrum analysis showed that Ag was distributed uniformly in the CNT/HA matrix. The breakage of CNTs into spheroid particles at higher temperatures (1700 degrees C) is attributed to the Rayleigh instability criterion. Mechanical properties (hardness and elastic modulus) of the samples were evaluated using nanoindentation testing. Ag reinforcement resulted in the enhancement of hardness (by similar to 15%) and elastic modulus (similar to 5%) of HA samples, whereas Ag reinforcement in CNT, Ag addition does not have much effect on hardness (0.3 GPa) and elastic modulus (5 GPa). The antibacterial tests performed using Escherichia coli and Staphylococcus epidermidis showed significant decrease (by similar to 65-86%) in the number of adhered bacteria in HA/CNT composites reinforced with 5% Ag nanoparticles. Thus, Ag-reinforced HA/CNT can serve as potential antibacterial biocomposites.

Twinning induced enhancement of fracture toughness in ultrafine grained Hydroxyapatite-Calcium Titanate composites

Despite being highly bioactive and biocompatible, the limitations of monolithic hydroxyapatite (HA) include extremely low fracture toughness, poor electrical conductivity. While addressing these issues, the present study demonstrates how CaTiO3 (CT) addition to HA can be utilized to obtain a combination of long crack fracture toughness (1.7 MPa m(1/2) SEVNB technique) and flexural strength of 98-155 MPa (3-point bending) and a moderate tensile strength (diametral compression) of 17-36 MPa. The enhancement in fracture resistance in spark plasma sintered HA-CT composites has been explained in reference to the observed twin morphology. TEM reveals the presence of twins in CT grains due to 1800 rotation about 101]. The measured properties along with our earlier reports on biocompatibility and electrical properties make HA-CT suitable for bone tissue engineering applications. When compared with other competing HA-based biocomposites, HA-CT composites are found to have a better combination of properties useful for medium load bearing implant applications. (C) 2015 Elsevier Ltd. All rights reserved.