In vitro characterization of laser ablation pseudowollastonite coating

Pseudowollastonite (CaSiO₃) coatings on titanium alloy substrates were prepared by laser ablation. The in vitro bioactivity of the coatings was examined for its biomedical applicability which was evaluated by immersion in human parotid saliva. The pseudowollastonite-coatings were soaked for various periods and characterized by SEM-EDS, XRD, FTIR, and TEM analysis, and the results indicated that the carbonated hydroxyapatite (CHA) was formed on the surface of the coatings within 1 day. In addition, cell attachment test showed that the pseudowollastonite-coatings supported the mesenchymal stem cells adhesion and spreading, and the cells established close contacts with the ceramics after 1 day of culture. These findings indicate that the pseudowollastonite-coatings possesses good bioactivity, biocompatibility and could be of interest in specific periodontal applications for bone restorative purposes.     

Development and cytotoxicity evaluation of SiAlONs ceramics

SiAlONs are ceramics with high potential as biomaterials due to their chemical stability, associated with suitable mechanical properties, such as high fracture toughness and fracture resistance. The objective of this work was to investigate the mechanical properties and the cytotoxicity of these ceramic materials. Three different compositions were prepared, using silicon nitride, aluminum nitride and a rare earth oxide mixture as starting powders, yielding Si₃N₄–SiAlON composites or pure SiAlON ceramics, after hot-pressing at 1750 °C, for 30 min. The sintered samples were characterized by X-ray diffraction analysis (XRD) and scanning electron microscopy (SEM). Furthermore, hardness and fracture toughness were determined using the Vicker's indentation method. The biological compatibility was evaluated by in vitro cytotoxicity tests. Ceramic with elevated hardness, ranging between 17 and 21 GPa, and high fracture toughness of 5 to 6 MPa m^1/2 were obtained. Since a nontoxic behavior was observed in the cytotoxicity tests, it may be assumed that SiAlON-based ceramics are viable materials for clinical applications.     

Fabrication and evaluation of physical properties and cytotoxicity of zein-based polyurethanes

Polyurethane prepolymer (PUP) was first synthesized from polycaprolactone diol and isophorone diisocyanate; and then a series of zein-based polyurethane (ZEPU) sheets was fabricated from PUP and zein (ZE) using a hot press and moulding process without addition of other additives. Effects of ZE content (W_ZE) on the structure and properties of the resultant ZEPU sheets were investigated by Fourier transform infrared spectroscopy, scanning electron microscopy, dynamic mechanical analysis, tensile testing, and dissolubility testing in alcohol. The results indicated that cross-linking and grafting reactions occurred between ZE and PUP to form new polyurethane showing a higher thermal stability, flexibility, and alcohol-resistance than the neat ZE sheets. For example, the elongation at break of ZEPU with 50 % W_ZE was 211.2 %, which was 47 times higher than that of neat ZE sheet. ZE molecules acted as both cross-linkers and polymer fillers in ZEPU sheets. The cytotoxicity and cytocompatibility of ZEPU sheets were evaluated by cell culture in vitro. The ZEPU sheets showed non- or low-cytotoxicity, and L929 cells grew and expanded well on the surfaces of the sheets with W_ZE over 50 %. Undoubtedly, the fabrication of ZE-based polyurethanes without toxic additives such as catalysts, cross-linkers and chain extenders improved the physical properties and cytocompatibility of zein, thus widening the possible range of applications for zein-based biomaterials.     

Stress protein assay for the evaluation of cytotoxicity of dental amalgam

To evaluate the cytotoxicity of mercury in dental amalgams, a stress protein assay was performed and the results were compared with the cytotoxicity evaluated by a neutral red uptake assay. The induction of a major stress protein, hsp70, was analyzed at levels of mRNA, synthesis and accumulation in human HeLa cells treated with extracts from amalgam, metal mercury and mercuric chloride. Mercuric chloride induced an increase in the synthesis of hsp70 at concentrations of mercury half those used for the neutral red uptake assay. The extracts from dental amalgam and metal mercury induced an increase in hsp70 mRNA at concentrations of mercury half those causing the inhibition of neutral red uptake into cells. Furthermore, the extracts from dental amalgam or metal mercury increased the synthesis of hsp70 and inhibited the uptake of dye at concentrations of mercury 1/10-1/50 lower than those at which mercuric chloride acted. These results suggest that the stress protein assay is more sensitive than the conventional neutral red assay for the evaluation of the cytotoxicity of mercury in dental amalgams and that the methods used in the preparation of metal solutions seem to be critical to the evaluation of cytotoxicity of dental materials.     

Preliminary evaluation of the in vitro cytotoxicity of PMMA-co-EHA bone cement

This work reports a preliminary in vitro cytotoxicity assessment of new poly (methyl methacrylate)-co-ethyl hexylacrylate (PMMA-co-EHA) bone cement by evaluating the effect of its leachables on the viability of human osteoblast-like cells (MG63 line) and their progression through the cell cycle. MG63 cells were exposed to 72 h-extract dilutions of PMMA-co-EHA and their viability was tested using the 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay. Also, putative changes in the progression of cells through the cell cycle were monitored using flow cytometry. For that the relative nuclear DNA content and the ratio of cells at G₁:S:G₂ stages of the cell cycle were measured after three exposure periods (24, 48 and 72 h). The obtained results revealed a dose-dependent influence of the cement extract in MG63 cell metabolism when compared to cells cultivated in a culture medium only. The MTT assay showed that a moderate number of cells died after exposure to the most concentrated extract. The cell cycle analysis revealed that leachables of PMMA-co-EHA led to significant changes in cellular proliferation, with cells exposed for 48 h to the most concentrated extract being arrested in the S phase of the cell cycle. However, despite the initial period of cytotoxicity, the obtained results suggest that after 72 h of exposure, the surviving cells are able to recover from this arresting condition and continue to proliferate. Therefore, this preliminary study indicates that, at the biological level, PMMA-co-EHA may have potential of being used as a bone cement matrix. However, a more detailed research work is needed to fully understand the factors responsible for the initial cytotoxicity observed.     

Indirect finite element evaluation of two-dimensional finite part integral using Fourier transformation

A broad class of engineering problems in fracture mechanics, thermal/fluid transport and electromagnetic theory involve the evaluation of two-dimensional finite part integrals of the form A method for evaluation of such integrals is developed by deriving an equivalent integral using Fourier transformation. This equivalent integral does not involve a kernel with singular behaviour. Consequently, standard numerical integration methodologies with conventional analytical evaluation techniques can be used in the finite element computations. The accuracy and convergence of the developed numerical procedure are successfully demonstrated by numerical examples for planar fracture geometries.     

熔体静电纺聚乳酸纤维膜的制备、表征及细胞毒性评价

调试熔体静电纺聚乳酸(PLA)过程中的电压场和温度场的参数,对不同条件下的纤维膜进行测试,研究电压场与温度场与纤维直径间的关系,并评价熔体静电纺PLA膜的细胞毒性。以聚乳酸(PLA)为原料,采用熔体静电纺丝方法,电压调整在20~26 k V范围,空间温度在10~70℃之间,分别进行熔体纺丝实验,将制得的纤维膜进行细胞毒性评价。熔体静电纺丝PLA纤维的平均直径随电压的升高逐渐增大,当空间温度为50℃时,所得纤维平均直径为最小。细胞活力测试证实熔体静电纺PLA膜无细胞毒性,具有良好的组织工程材料的应用前景。     

The cytotoxicity evaluation of Kevlar and silicon carbide by MTT assay

The MTT test has been widely used as a rapid and sensitive method for screening anticancer drugs. In this paper, we used this method to assess the cytocompatibility of three materials: Keviar 29, silicon carbide and polyvinyl chloride (PVC) in both a quantitative and a qualitative manner. The materials were prepared by cleaning in 70% ethanol, autoclaved or gamma-sterilized. Extracts were prepared at four time periods (1, 2, 3 and 4 weeks) and two temperatures (37°C and 80°C). The extracts were used in the MTT assay and the data were collected and analysed with ONEWAY and DUNCAN procedures using the statistical computer package SPSS^x. The MTT staining procedure was also used in direct contact with the materials. The result from the MTT assay demonstrated that Kevlar, SiC and PVC extracted at 37°C were not cytotoxic while PVC extracted at 80°C did show some cytotoxicity, especially the material that had been gamma-sterilized. In the direct contact test the Kevlar showed no cytotoxicity. The SiC did show some localized toxicity when the material had been autoclaved, however, SiC subject to prior cleaning with ethanol showed no cytotoxicity. The PVC that had been autoclaved caused a cytotoxic response whereas the material that had been gamma-sterilized or cleaned in ethanol showed good cytocompatibility. This paper demonstrates that the MTT staining procedure is a useful technique to study the cytocompatibility of materials in both a quantitative and a qualitative manner. It is also shown that the cellular response to the materials tested is dependent on the method of preparation.     

On-chip evaluation of shear stress effect on cytotoxicity of mesoporous silica nanoparticles

In this work, nanotoxicity in the bloodstream was modeled, and the cytotoxicity of sub-50 nm mesoporous silica nanoparticles to human endothelial cells was investigated under microfluidic flow conditions. Compared to traditional in vitro cytotoxicity assays performed under static conditions, unmodified mesoporous silica nanoparticles show higher and shear stress-dependent toxicity to endothelial cells under flow conditions. Interestingly, even under flow conditions, highly organo-modified mesoporous silica nanoparticles show no significant toxicity to endothelial cells. This paper clearly demonstrates that shear stress is an important factor to be considered in in vitro nanotoxicology assessments and provides a simple device for pursuing this consideration.     

Comparative cytotoxicity evaluation of different size gold nanoparticles in human dermal fibroblasts

The aim of this work was to compare the effects of three commercially available gold nanoparticles (AuNPs) of different sizes (30, 50 and 90 nm) on the viability of normal human dermal fibroblasts (NHDF). In addition, we evaluated protective effect of N-Acetyl-L-cysteine (NAC), total glutathione content (GSH/GSSG), superoxide dismutase (SOD) activity and reactive oxygen species (ROS) production to investigate if oxidative stress was involved in the cytotoxic response of these AuNPs. Although AuNP-induced cytotoxicity was dose and time dependent, nanoparticle size slightly influenced the cytotoxic response of AuNPs assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and lactate dehydrogenase. Regarding oxidative parameters, NAC produced no significant protection of NHDF cells against treatment with any of the three AuNPs. Independently on nanoparticle size, GSH/GSSG content was drastically depleted after 24 h of incubation with the three AuNPs (less than 15% in all cases), while no statistically significant changes on SOD activity were reported (～90% of activity). The three AuNPs also caused a notable increase in the ROS production of NHDF cells. In conclusion, our data suggest that AuNP-induced cytotoxicity in NHDF is mediated by oxidative stress and it is independent of nanoparticle size.     

A semi-automated cell culture evaluation system for cytotoxicity testing of dental materials

A rapid and quantitative determination of viable cells in the cytotoxicity testing of dental materials is desirable to evaluate large numbers of samples in a short time. For this purpose a new and semi-automated cell culture evaluation system was developed using the fluorogenic dye fluorescein diacetate (FDA) for vital staining and microtiter plates for culture vessels. Our methodological experiments showed that in this system the fluorescence intensity was linearily related to the cell number from 500 to 20 000 cells per culture^–1 and that fluorescence recording was stable for between at least 1 to 2 h using a quencher solution for absorbing extracellular fluorescence. The results from toxicity testing of different dental materials (two glass ionomer cements, a phosphate cement, a composite resin and monomeric methylmethacrylate) corresponded to those derived from other, standard test methods. Because of the ease of performance, the quantitative, rapid evaluation system and the small culture vessels requiring only few cells per culture, the test method presented may be an interesting alternative to other cell culture techniques for cytotoxicity testing.     

Tamoxifen-encapsulated vesicular systems: cytotoxicity evaluation in human epidermal keratinocyte cell line

Aim: Tamoxifen is a nonsteroidal estrogen receptor modulator indicated in the treatment of breast cancer. Apoptosis has been reported to be a major mechanism for its antitumor effect. Tamoxifen has also shown significant potential in treating various dermatological disorders including psoriasis, characterized by hyperproliferation of epidermal keratinocytes. An endeavor was made in the current studies to investigate the potency of vesicle-encapsulated tamoxifen on human epidermal keratinocyte cell lines. Methods: Drug was encapsulated in the phospholipid-based vesicular systems, namely, conventional liposomes and flexible-membrane liposomes. In vitro cytotoxicity evaluation of the formulations was carried out employing MTT cell proliferation assay. Results: A composition-dependent strong inhibition in the viability of epidermal keratinocyte cells was observed. Conclusion: The encouraging findings of this work construe immense potential of the tamoxifen-encapsulated vesicular systems in the management of psoriasis.     

Indirect measurements

Conclusions The given relationship between the measured quantities as defined in the classifications [1, 2], or in the measurement equation [3] should be interpreted as a statistical relationship (with a determinate relationship constituting a particular case), because it is necessary to bear in mind the completeness of measurement classifications.Translated from Izmeritel'naya Tekhnika, No. 10, pp. 83–84, October, 1973.     

Corrosion performances in simulated body fluids and cytotoxicity evaluation of Fe-based bulk metallic glasses

The aim of this work is to investigate the corrosion resistance and biocompatibility of three kinds of Fe based bulk metallic glasses (BMGs), Fe₄₁Co₇Cr₁₅Mo₁₄C₁₅B₆Y₂ (BMG1), (Fe₄₄Cr₅Co₅Mo₁₃Mn₁₁C₁₆B₆)₉₈Y₂ (BMG2), and Fe₄₈Cr₁₅Mo₁₄C₁₅B₆Er₂ (BMG3) by electrochemical measurements and indirect contact cytotoxicity assays, respectively. In comparison with 316 L SS biomedical steel, Fe based BMGs show better corrosion resistance in both simulated body fluids (Hank's solution and artificial saliva). The OCP curves show that the passive film on the Fe based BMG surfaces is quite stable, like 316 L SS. The corrosion current densities obtained from the anodic polarization curves from the lowest to highest are as follows: BMG3 < BMG1 < BMG2 < 316 L SS. The EIS analysis indicates that the Fe Based BMGs have larger polarization resistance value than that of 316 L SS except for BMG2 in artificial saliva. The pitting corrosion potentials of Fe based BMGs are much higher than that of the 316 L SS, resulting in very few ions releasing into the electrolytes while a significant amount of Ni and Fe ions release was found for 316 L SS under the same condition. The indirect cytotoxicity results suggest that all three Fe based BMG extracts have no cytotoxicity to L929 and NIH3T3 cells. All these results demonstrate that Fe based BMGs will open up a new path for the biomedical applications, especially in dental implantology.     

In vitro safety toxicology data for evaluation of gold nanoparticles-chronic cytotoxicity, genotoxicity and uptake

Safety and toxic effects of nanoparticles are still largely unexplored due to the multiple aspects that influence their behaviour toward biological systems. Here, we focus the attention on 12 nm spherical gold nanoparticle coated or not with hyaluronic acid compared to its precursor counterpart salt. Results ranging from the effects of a 10-days exposure in an in vitro model with BALB/c 3T3 fibroblast cells show how 12 nm spherical gold nanoparticles are internalized from 3T3 cells by endo-lysosomal pathway by an indirect measurement technique; and how gold nanoparticles, though not being a severe cytotoxicant, induce DNA damage probably through an indirect mechanism due to oxidative stress. While coating them with hyaluronic acid reduces gold nanoparticles cytotoxicity and slows their cell internalization. These results will be of great interest to medicine, since they indicate that gold nanoparticles (with or without coating) are suitable for therapeutic applications due to their tunable cell uptake and low toxicity.     

Indirect patent citations

Patent citations are extensively used as a measure of patent quality. However, counting citations does not account for the fact that citations come from patents of different qualities, and that citations are of variable qualities. We develop a citation index which takes into account the cumulative quality of the citing patents. We apply this index to the 2,139,314 utility patents granted in the U.S. between 1975 and 1999. We study the properties of this index by year and by technological category, and analyse the links between patents.     

The in vitro indirect cytotoxicity test and in vivo interface bioactivity evaluation of biodegradable FHA coated Mg-Zn alloys

A kind of biodegradable fluoridated hydroxyapatite (FHA) coating was prepared on Mg-Zn alloy to improve the interface bioactivity in bone healing via electrodeposition method. The in vitro cytotoxicity evaluation of the ions released during degradation was taken. No toxicity was shown and even higher cells’ viability appeared on the 7th day compared with the normal culture case (negative control). In vivo implantation was carried out in the femoral condyle of adult New Zealand rabbits. The cross section showed by Micro-CT scan confirmed that the better interface contacts happened in the coated group after one month implantation. Also the coating left can still be normally observed by scanning electron microscope (SEM) with a little degradation. As a result, the FHA coating may be a promising candidate to enhance interface bioactivity for biodegradable Mg alloys in orthopaedics.     

Indirect method of checking conveyor scales

Translated from Izmeritel'naya Tekhnika, No. 5, pp. 18–20, May, 1992.