Silicone based polyurethane materials: a promising biocompatible elastomeric formulation for cardiovascular applications

The biocompatibility of a new material for cardiovascular applications constituted by a poly(ether)urethane (PEtU) and a silicone [polydimethylsiloxane (PDMS)] was evaluated. The achieved material shows properties similar to both polyurethanes and silicones. The material was transformed into porous membranes by a spray-deposition technique. Since any material preparation and manufacturing procedure may introduce some toxicity, in vitro cytotoxicity screening tests were carried out. Human umbilical vein endothelial cells (HUVECs) and a mouse fibroblasts cell line (L929) were cultivated with extracts obtained from materials containing 10, 40 and 100% (w/w) of PDMS. The commercially available Estane 5714-F1 and Cardiothane 51 were used as controls. Extracts were incubated up to 72 hours with HUVECs and L929 cells. The cytotoxic effect was evaluated by light microscopy, cell viability (MTT reduction and neutral red uptake) and proliferation (5-bromo-2'-deoxyuridine incorporation) tests. In vivo studies were carried out using materials containing the same PDMS percentages as for in vitro experiments. The same commercial controls were used. Results obtained with cell culture studies agreed with those obtained in the in vivo experiments and showed that the material preparation and manufacturing procedure do not introduce any toxicity in the products at each PDMS concentration investigated.     

Toxicity Assessments of Multisized Gold and Silver Nanoparticles in Zebrafish Embryos

The potential toxicity of nanoparticles is addressed by utilizing a putative attractive model in developmental biology and genetics: the zebrafish (Danio rerio). Transparent zebrafish embryos, possessing a high degree of homology to the human genome, offer an economically feasible, medium‐througput screening platform for noninvasive real‐time assessments of toxicity. Using colloidal silver (cAg) and gold nanoparticles (cAu) in a panoply of sizes (3, 10, 50, and 100 nm) and a semiquantitative scoring system, it is found that cAg produces almost 100% mortality at 120 h post‐fertilization, while cAu produces less than 3% mortality at the same time point. Furthermore, while cAu induces minimal sublethal toxic effects, cAg treatments generate a variety of embryonic morphological malformations. Both cAg and cAu are taken up by the embryos and control experiments, suggesting that cAg toxicity is caused by the nanoparticles themselves or Ag⁺ that is formed during in vivo nanoparticle destabilization. Although cAg toxicity is slightly size dependent at certain concentrations and time points, the most striking result is that parallel sizes of cAg and cAu induce significantly different toxic profiles, with the former being toxic and the latter being inert in all exposed sizes. Therefore, it is proposed that nanoparticle chemistry is as, if not more, important than specific nanosizes at inducing toxicity in vivo. Ultimately such assessments using the zebrafish embryo model should lead to the identification of nanomaterial characteristics that afford minimal or no toxicity and guide more rational designs of materials on the nanoscale.     

Ocular permeability of pirenzepine hydrochloride enhanced by methoxy poly(ethylene glycol)-poly(D, L-lactide) block copolymer

Methoxy poly(ethylene glycol)-poly(D,L-lactide) block copolymer was tested as an ocular permeation enhancer for pirenzepine hydrochloride. The block copolymers with the methoxy poly(ethylene glycol) to poly(D,L-lactide) weight ratio of 80/20, 50/50, 40/60 were synthesized by a ring-opening polymerization procedure. In vitro transcorneal experiments demonstrated that the block copolymer 80/20 significantly enhanced the transcorneal permeation of pirenzepine at the mass ratio of 1/1.4 (pirenzepine hydrochloride/copolymer). Interaction between pirenzepine and copolymer was identified by infrared spectroscopy analysis and dialysis experiments. Ocular pharmacokinetics of pirenzepine/copolymer preparation by in vivo instillation experiments confirmed that block copolymer could enhance the ocular penetration of pirenzepine. Ocular chronic toxicity experiments of block copolymer and pirenzepine/copolymer preparation were studied on rabbits, and no significant toxicity in both groups was observed within 9 months. It could conclude that pirenzepine/copolymer preparation is effective and safe in ocular delivery of pirenzepine.     

Ocular Permeability of Pirenzepine Hydrochloride Enhanced by Methoxy poly(ethylene glycol)–poly(D,L-lactide) Block Copolymer

Methoxy poly(ethylene glycol)–poly(D,L-lactide) block copolymer was tested as an ocular permeation enhancer for pirenzepine hydrochloride. The block copolymers with the methoxy poly(ethylene glycol) to poly(D,L-lactide) weight ratio of 80/20, 50/50, 40/60 were synthesized by a ring-opening polymerization procedure. In vitro transcorneal experiments demonstrated that the block copolymer 80/20 significantly enhanced the transcorneal permeation of pirenzepine at the mass ratio of 1/1.4 (pirenzepine hydrochloride/copolymer). Interaction between pirenzepine and copolymer was identified by infrared spectroscopy analysis and dialysis experiments. Ocular pharmacokinetics of pirenzepine/copolymer preparation by in vivo instillation experiments confirmed that block copolymer could enhance the ocular penetration of pirenzepine. Ocular chronic toxicity experiments of block copolymer and pirenzepine/copolymer preparation were studied on rabbits, and no significant toxicity in both groups was observed within 9 months. It could conclude that pirenzepine/copolymer preparation is effective and safe in ocular delivery of pirenzepine.     

Shape-dependent cytotoxicity of polyaniline nanomaterials in human fibroblast cells

The toxicity of polyaniline (PANI) nanomaterials with four different aspect ratios on human lung fibroblast cells was investigated by cell viability assay, cytotoxicity assay, apoptosis/necrosis measurement, and reactive oxygen species production. The toxicity increased with decreasing aspect ratio of PANI nanomaterials. In contrast, the highest aspect ratio PANI nanomaterials showed similar results with bulk PANI materials. The adverse effect of PANI nanomaterials was also concentration- and time-dependent. Low aspect ratio PANI nanomaterials induced more necrosis and more reactive oxygen species than others. These results provide new understanding of shape-dependent toxicity of nanomaterials.     

Genotoxicity tests of poly(styrene-co-maleic anhydride)-coated silver nanoparticles in vivo and in vitro

Nanosilver is thought to hold potential for use in medical materials. The safety of newly developed poly(styrene-co-maleic anhydride)-coated silver nanoparticles (SMA-AgNPs) requires investigation. In this study, three in vitro and in vivo experiments for investigating genetic toxicity–the Ames test, a micronucleus assay, and a chromosome aberration test–were conducted. Results from the Ames testing showed SMA-AgNPs to have a negative effect, either with or without S9 metabolism. In addition, SMA-AgNPs increased the number of reticulocytes and micronuclei in reticulocytes at 48 and 72 h after treatment. Indeed, SMA-AgNPs induced significant changes in the chromosomal aberration rate in CHO-K1 (Chinese hamster ovary cell clone K1) cells. In conclusion, SMA-AgNPs did cause DNA damage in terms of chromosomal aberration and may have a potential genotoxic effect in certain applications.     

Cellular and in vitro toxicity of nanodiamond-polyaniline composites in mammalian and bacterial cell

The composite nanodiamond (ND) particles are rapidly emerging as promising material for the next generation agent for drug delivery, biosensors, or imaging contrast applications. Consequently, the health risks associated with the exposure to nanocomposite materials are utmost important. The objective of our work is to study the toxic effects of the nanodiamond-polyaniline (ND-Pani) composites in Human Embryonic Kidney (HEK) cell line. Toxic effects of either ND or ND-Pani nanocomposite in powder form on HEK293 are tested using MTT assay. Results indicated that there was no significant difference in the cell survival between samples treated with ND-Pani nanocomposite and the control sample for the two lower concentrations used in the experiments (p < 0.05). Morphology of the cells was not significantly affected due to the inclusion of nanocomposites during the incubation phase. The stability of the film was also improved due to the inclusion of the nanodiamond particles into the polymeric matrix making it suitable for electrochemical sensing applications. Experimental results have shown that the toxicity effect of lower concentration of ND-Pani composite is negligible, thus indicating that below 1 μg/ml could be a safer range without secondary toxicity effect.     

In Vivo NIR Fluorescence Imaging,Biodistribution, and Toxicology of Photoluminescent Carbon Dots Produced from Carbon Nanotubes and Graphite

Oxidization of carbon nanotubes by a mixed acid has been utilized as a standard method to functionalize carbon nanomaterials for years. Here, the products obtained from carbon nanotubes and graphite after a mixed‐acid treatment are carefully studied. Nearly identical carbon dot (Cdot) products with diameters of 3–4 nm are produced using this approach from a variety of carbon starting materials, including single‐walled carbon nanotubes, multiwalled carbon nanotubes, and graphite. These Cdots exhibit strong yellow fluorescence under UV irradiation and shifted emission peaks as the excitation wavelength is changed. In vivo fluorescence imaging with Cdots is then demonstrated in mouse experiments, by using varied excitation wavelengths including some in the near‐infrared (NIR) region. Furthermore, in vivo biodistribution and toxicology of those Cdots in mice over different periods of time are studied; no noticeable signs of toxicity for Cdots to the treated animals are discovered. This work provides a facile method to synthesize Cdots as safe non‐heavy‐metal‐containing fluorescent nanoprobes, promising for applications in biomedical imaging.     

Semiconductor quantum dots for in vivo imaging

Quantum dots play an important role in the in vitro, ex vivo, and in vivo optical imaging. Dramatic improvements have been achieved in the aspect of surface modification, biocompatibility, and targeting specificity, which had significant impact on the in vivo applications of quantum dots. This review summarizes the recent advances of quantum dots for in vivo imaging using both non-specific and targeted approaches. The toxicity of cadmium chalcogenide materials and alternative approaches such as the use of doped nanocrystal quantum dots were also discussed. The integration of quantum dots with other imaging techniques is also expected to give rise to a new generation of multifunctional probes for biomedical applications.     

Synthesis and characterization of multifunctional iron oxide nanoparticles

In order to get high water solubility, monodisperse, superparamagnetic nanoparticles, poly (acrylic acid) was employed to modify Fe3O4 by a high-temperature solution-phase hydrolysis approach. Then, folic acid (FA) and fluorescein isothiocyanate were successively conjugated with prepared magnetic nanoparticles (MNPs). The functional MNPs were characterized by X-ray diffraction (XRD), dynamic light scattering (DLS), transmission electron microscope (TEM), inductively coupled plasma-atomic emission spectrometer (ICP-AES), and vibrating sample magnetometer (VSM), respectively. The toxicity of the materials was evaluated by selecting NIH/3T3 fibroblast cells and no toxic effect was observed. The fluorescent imaging and targeting property of the MNPs were also realized in vitro and in vivo experiments by confocal laser scanning microscopy (CLSM) and Kodak In-Vivo FX Professional Imaging System, respectively. The results indicated that the final products exhibited interesting magnetic, optical and targeting properties for further potential applications in biological and biomedical fields.     

The response of primary rat and human osteoblasts and an immortalized rat osteoblast cell line to orthopaedic materials: comparative sensitivity of several toxicity indices

When studying the biocompatibility of orthopaedic biomaterials it isoften necessary to discriminate between responses which show mild cytotoxicity.It is therefore essential to use a very sensitive index of toxicity. We havecompared the sensitivity of four well-established indices of toxicity: totalcell protein content, leakage of lactate dehydrogenase (LDH), reducedglutathione content and the MTT assay, with that of a novel index, alkalinephosphatase (ALP) activity. Comparisons were made by detecting nickel chloridetoxicity in osteoblasts. ALP activity, the novel method, proved the mostsensitive index of toxicity and it provides a convenient automated assay forassessing the interactions of materials with osteoblasts. The responses tonickel chloride and to aqueous extracts prepared from carbon fibre reinforcedepoxy and polyetheretherketone (peek), two candidate materials for orthopaedicimplants, were compared in primary and immortalized rat osteoblasts, and !in primary human osteoblasts. Although the immortalized rat osteoblast cell line,FFC, was consistently the most sensitive cell type, the responses of the humancells and the FFC cell line were similar in terms of ALP activity throughout therange of nickel concentrations studied. Neither peek nor epoxy material extractsshowed a significant decrease in the MTT or ALP responses in any of the threecell types. Our data suggest that immortalized rat osteoblasts may provide anin vitro model system for screening the biocompatibility of orthopaedicpolymers.     

In vitro and in vivo toxicity of CdTe nanoparticles

Cadmium telluride (CdTe) nanoparticles exhibit strong and stable fluorescence that is attractive for many applications such as biological probing and solid state lighting. The evaluation of nanoparticle toxicity is important for realizing these practical applications. However, no systematic studies of CdTe nanoparticle toxicity have been reported. We investigated and compared the size- and concentration-dependent cytotoxicity of CdTe nanoparticles in human hepatoma HepG2 cells using the MTT assay. CdTe nanoparticles elicited cytotoxicity in a concentration- and size-dependent manner, with smaller-sized particles exhibiting somewhat higher potency. Lesser cytotoxicity of partially purified CdTe-Red particles (following methanol precipitation and resuspension) suggested that free cadmium ions may contribute to cytotoxicity. We also evaluated the acute toxicity of CdTe-Red particles following intravenous exposure in male rats (2 micromol/kg). Few signs of functional toxicity or clinical (urinary or blood) changes were noted. Interestingly, motor activity was transiently reduced (2 hours after treatment) and then significantly increased at a later timepoint (24 hours after dosing). These studies provide a framework for further characterizing the in vitro and in vivo toxic potential of different types of CdTe nanoparticles and suggest that the nervous system may be targeted by these nanoparticles under some conditions.     

Safety assessment of nanoparamagnetic contrast agents with different coatings for molecular MRI

Despite the wide application of gadolinium as a contrast agent for magnetic resonance imaging (MRI), there is a serious lack of information on its toxicity. Gadolinium and gadolinium oxide (Gd-oxide) are used as contrast agents for magnetic resonance imaging (MRI). There are methods for reducing toxicity of these materials, such as core nanoparticles coating or conjugating. Therefore, for toxicity evaluation, we compared the viability of commercial contrast agents in MRI (Gd-DTPA) and three nanoparticles with the same core Gd₂O₃ and small particulate gadolinium oxide or SPGO (< 40 nm) but different coatings of diethyleneglycol (DEG) as Gd₂O₃-DEG and methoxy polyethylene glycol-silane (mPEG-silane: 550 and 2000 Dalton) as SPGO-mPEG-silane550 and SPGO-mPEG-silane2000, respectively, in the SK-MEL3 cell line, by light microscopy, MTT assay using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide, and the LDH assay detecting lactate dehydrogenase activity. The viability values were not statistically different between the three nanoparticles and Gd-DTPA. The MTT and LDH assay results showed that Gd₂O₃-DEG nanoparticles were more toxic than Gd-DTPA and other nanoparticles. Also, SPGO-mPEG-silane2000 was more biocompatible than other nanoparticles. The obtained results did not show any significant increase in cytotoxicity of the nanoparticles and Gd-DTPA, neither dose-dependent nor time-dependent. Therefore, DEG and PEG, due to their considerable properties and irregular sizes (different molecular weights), were selected as the useful surface covering materials of nanomagnetic particles that could reveal noticeable relaxivity and biocompatibility characteristics.     

On the Immersion Testing of Degradable Implant Materials in Simulated Body Fluid: Active pH Regulation Using CO2

Predictions of in vivo degradation behavior derived from laboratory experiments are of great importance in the development of biodegradable materials. A key issue is the simulation of the physiological conditions found in living organisms. Generally, the testing solution and in particular the pH buffer have a significant influence on the outcome of degradation experiments. This study presents results obtained from immersion tests in SBF buffered with gaseous CO₂. The control of the pH value by means of CO₂ has the advantage of better reflecting physiological conditions, and allows the carrying out of in vitro experiments which are closer to the in vivo situation.     

C60 fullerene enhances cisplatin anticancer activity and overcomes tumor cell drug resistance

We formulated and analyzed a novel nanoformulation of the anticancer drug cisplatin (Cis) with C60 fullerene (C60+Cis complex) and showed its higher toxicity toward tumor cell lines in vitro when compared to Cis alone.The highest toxicity of the complex was observed in HL-60/adr and HL-60/vinc chemotherapyresistant human leukemia cell sublines (resistant to Adriamycin and Vinculin,respectively).We discovered that the action of the C60+Cis complex is associated with overcoming the drug resistance of the tumor cell lines through observing an increased number of apoptotic cells in the Annexin V/PI assay.Moreover,in vivo assays with Lewis lung carcinoma (LLC) C57BL/6J male mice showed that the C60+Cis complex increases tumor growth inhibition,when compared to Cis or C60 fullerenes alone.Simultaneous1y,we conducted a molecular docking study and performed an Ames test.Molecular docking specifies the capability of a C60 fullerene to form van der Waals interactions with potential binding sites on P-glycoprotein (P-gp),multidrug resistance protein 1 (MRP-1),and multidrug resistance protein 2 (MRP-2) molecules.The observed phenomenon revealed a possible mechanism to bypass tumor cell drug resistance by the C,o+Cis complex.Additionally,the results of the Ames test show that the formation of such a complex diminishes the Cis mutagenic activity and may reduce the probability of secondary neoplasm formation.In conclusion,the C60+Cis complex effectively induced tumor cell death in vitro and inhibited tumor growth in vivo,overcoming drug resistance likely by the potential of the C60 fullerene to interact with P-gp,MRP-1,and MRP-2 molecules.Thus,the C60+Cis complex might be a potential novel chemotherapy modification.     

Immobilization of type-I collagen and basic fibroblast growth factor (bFGF) onto poly (HEMA-co-MMA) hydrogel surface and its cytotoxicity study

Type-I collagen and bFGF were immobilized onto the surface of poly (HEMA-co-MMA) hydrogel by grafting and coating methods to improve its cytotoxicity. The multi-layered structure of the biocompatible layer was confirmed by FTIR, AFM and static water contact angles. The layers were stable in body-like environment (pH 7.4). Human skin fibroblast cells (HSFC) were seeded onto Col/bFGF-poly (HEMA-co-MMA), Col-poly (HEMA-co-MMA) and poly (HEMA-co-MMA) films for 1, 3 and 5 day. MTT assay was performed to evaluate the extraction toxicity of the materials. Results showed that the cell attachment, proliferation and differentiation on Col/bFGF-poly (HEMA-co-MMA) film were higher than those of the control group, which indicated the improvement of cell-material interaction. The extraction toxicity of the modified materials was also lower than that of the unmodified group. The protein and bFGF immobilized poly (HEMA-co-MMA) hydrogel might hold great promise to be a biocompatible material.     

超细大气矿物颗粒物界面反应及生物活性研究新进展

矿物粉尘是大气颗粒物的重要组成部分,与呼吸系统和心血管系统空气污染疾病密切相关。纳米颗粒物由于微尺度效应、高比表面积和复杂的化学构成具有与常规物质不同的活性。通过研究超细大气颗粒物的表面电性、表面基团、矿物-生物化学作用、体外生物溶解界面反应及自由基等方面的运动与沉降颗粒物的有机-无机体系的界面反应机理及其生物活性的现状与方向,进而指出应从微观体系界面反应的模拟和实验等角度重点揭示其化学-生物活性和毒性,以多学科联合方式对天然超细矿物颗粒物的生物活性进行深入研究。     

Preparation of an electrodeposited hydroxyapatite coating on titanium substrate suitable for in-vivo applications

In this paper the porous hydroxyapatite coating on Ti implant materials was prepared by the process of electrodeposition, hydrothermal and sinter. The surface morphology, bond strength and thickness of HA coatings were investigated by SEM, AFM, and its biocompatibility was evaluated by cytotoxicity experiments and implant experiments, respectively. Results showed that (1) The HA coatings was 50 μm thickness and adhered on the Ti substrate strongly, which bond strength reached 38MPa. AFM analysis showed that the HA coating was porous structure, in which the mean pore size was 236.5 μm, (2) Cytotoxicity experiments and implant experiments showed that HA-coated Ti implant materials has little cytotoxicity in vitro and little inflammatory reaction in vivo, and there were no statistically disparity between HA-coated Ti implant and titanium implant materials of clinical application (p > 0.05), which demonstrated that HA-coated Ti has a good biocompatibility.     

Cytotoxicity of graphene oxide and graphene in human erythrocytes and skin fibroblasts

Two-dimensional carbon-based nanomaterials, including graphene oxide and graphene, are potential candidates for biomedical applications such as sensors, cell labeling, bacterial inhibition, and drug delivery. Herein, we explore the biocompatibility of graphene-related materials with controlled physical and chemical properties. The size and extent of exfoliation of graphene oxide sheets was varied by sonication intensity and time. Graphene sheets were obtained from graphene oxide by a simple (hydrazine-free) hydrothermal route. The particle size, morphology, exfoliation extent, oxygen content, and surface charge of graphene oxide and graphene were characterized by wide-angle powder X-ray diffraction, atomic force microscopy, X-ray photoelectron spectroscopy, dynamic light scattering, and zeta-potential. One method of toxicity assessment was based on measurement of the efflux of hemoglobin from suspended red blood cells. At the smallest size, graphene oxide showed the greatest hemolytic activity, whereas aggregated graphene sheets exhibited the lowest hemolytic activity. Coating graphene oxide with chitosan nearly eliminated hemolytic activity. Together, these results demonstrate that particle size, particulate state, and oxygen content/surface charge of graphene have a strong impact on biological/toxicological responses to red blood cells. In addition, the cytotoxicity of graphene oxide and graphene sheets was investigated by measuring mitochondrial activity in adherent human skin fibroblasts using two assays. The methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay, a typical nanotoxicity assay, fails to predict the toxicity of graphene oxide and graphene toxicity because of the spontaneous reduction of MTT by graphene and graphene oxide, resulting in a false positive signal. However, appropriate alternate assessments, using the water-soluble tetrazolium salt (WST-8), trypan blue exclusion, and reactive oxygen species assay reveal that the compacted graphene sheets are more damaging to mammalian fibroblasts than the less densely packed graphene oxide. Clearly, the toxicity of graphene and graphene oxide depends on the exposure environment (i.e., whether or not aggregation occurs) and mode of interaction with cells (i.e., suspension versus adherent cell types).     

In vitro and in vivo biocompatibility of graded hydroxyapatite–zirconia composite bioceramic

To obtain bioceramics with good osteoinductive ability and mechanical strength, graded hydroxyapatite–zirconia (HA–ZrO₂) composite bioceramics were prepared in this work. The biocompatibility of the bioceramics was investigated in vitro based on acute toxicity and cytotoxicity tests and hemolysis assay. Results showed the studied graded HA–ZrO₂ had little toxicity to mouse and L929 mouse fibroblasts. Also, hemolysis assay indicated a good blood compatibility of the bioceramics. Based on the results of in vitro tests, animal experiments were performed on white New Zealand rabbits by implantation into hip muscles and femur. It was found that the graded HA–ZrO₂ composite bioceramics exhibited superior osteoinductive ability, which may be a promising bioceramics implant.