In Vitro Cytotoxicity of Nanoparticles in Mammalian Germ-Line Stem Cell

Nanotechnology involves the creation and manipulation of materialsat nanoscale levels to create products that exhibit novel properties.There are important applications of nanoscience in biology andbiotechnology, and nanotechnology offers new tools to biologists(Whitesides, 2003). Nevertheless, despite the increased interest in the developmentof nanoparticles, few studies address their potential toxicity.The rapidly developing field of nanotechnology is likely tobecome yet another source of human exposure to nanoparticlesby different routes: inhalation, ingestion, dermal, and injection.Regulatory agencies, researchers, and health and environmentalwatchdogs are assessing how nanoscale materials affect humanhealth and the environment (Service, 2004). Similarly, the characteristic biokinetic behavior of nanoparticles     

Synthesis,Cytotoxicity and In Vitro Antileishmanial Activity of Naphthothiazoles

The leishmaniasis is a spectral disease caused by the protozoan Leishmania spp., which threatens millions of people worldwide. Current treatments exhibit high toxicity, and there is no vaccine available. The need for new lead compounds with leishmanicidal activity is urgent. Considering that many lead leishmanicidal compounds contain a quinoidal scaffold and the thiazole heterocyclic ring is found in a number of antimicrobial drugs, we proposed a hybridization approach to generate a diverse set of semi‐synthetic heterocycles with antileishmanial activity. We found that almost all synthesized compounds demonstrated potent activity against promastigotes of Leishmania (Viannia) braziliensis and reduced the survival index of Leishmania amastigotes in mammalian macrophages. Furthermore, the compounds were not cytotoxic to macrophages at fivefold higher concentrations than the EC₅₀ for promastigotes. All molecules fulfilled Lipinski's Rule of Five, which predicts efficient orally absorption and permeation through biological membranes, the in silico pharmacokinetic profile confirmed these characteristics. The potent and selective activity of semi‐synthetic naphthothiazoles against promastigotes and amastigotes reveals that the 2‐amino‐naphthothiazole ring may represent a scaffold for the design of compounds with leishmanicidal properties and encourage the development of drug formulation and new compounds for further studies in vivo.     

不同来源一次性使用无菌注射器用活塞体外细胞毒性的比较研究

目的：对不同来源一次性使用无菌注射器用活塞体外细胞毒性进行评价,为科学监管提供技术支持。方法：将一次性使用无菌注射器用活塞浸提液分别与L929细胞接触培养,采用四唑盐比色法（MTT法）量化细胞毒性,计算相对增殖率,并进行细胞毒性评价。结果：不同厂家相同浸提比例,产品之间细胞毒性均存在差异,同一厂家不同浸提比例的条件下,产品细胞毒性未发现明显变化。结论：不同厂家产品配方及生产工艺不尽相同可能是导致不同厂家的产品的细胞毒性存在差异的原因,建议在产品标准中生物性能技术要求中增加细胞毒性评价项。     

Cytotoxicity of Lidocaine to Human Corneal Endothelial Cells In Vitro

Lidocaine has been reported to induce apoptosis on rabbit corneal endothelial cells. However, the apoptotic effect and exact mechanism involved in cytotoxicity of lidocaine are not well‐established in human corneal endothelial (HCE) cells. In this study, we investigated the apoptosis‐inducing effect of lidocaine on HCE cells in vitro. After HCE cells were treated with lidocaine at concentrations of 0.15625–10.0 g/l, the morphology and ultrastructure of the cells were observed by inverted light microscope and transmission electron microscope (TEM). Cell viability was measured by MTT assay, and the apoptotic ratio was evaluated with flow cytometry and fluorescent microscopic counting after FITC–Annexin V/PI and AO/EB staining. DNA fragmentation was detected by electrophoresis, and the activation of caspases was evaluated by ELISA. In addition, changes in mitochondrial membrane potential were determined by JC‐1 staining. Results suggest that lidocaine above 1.25 g/l reduced cellular viability and triggered apoptosis in HCE cells in a time‐ and dose‐dependent manner. Diminishment of ΔΨm and the activation of caspases indicate that lidocaine‐induced apoptosis was caspase dependent and may be related to mitochondrial pathway.     

Potentiation of Cytotoxicity of Mitoxantrone Toward CHO-K1 Cells in Vitro by Dipyridamole

Dipyridamole (DP), a clinically used vasodilator and an antiplatelet compound, augmented the activity of the anticancer drug mitoxantrone (MXN) toward Chinese hamster ovary (CHO-K1) cells in culture. Clonogenic assays indicated that DP (1.0, 2.5, and 5.0 µM) decreased the survival of cells treated with MXN (5 to 25 nM) in a dose-dependent manner. Further, DP (1 and 5 nM) decreased the MXN concentration required for 50% inhibition of cell growth from 3.2 to 1.8 and from 3.0 to 0.5 nM, respectively, over a period of 3 days. DP (10 µM) increased the accumulation of MXN by 1.8-fold in exponentially growing cells exposed to MXN. The enhanced levels of MXN in CHO-K1 cells in the presence of the chemosensitizer may account for the potentiation of MXN-cytotoxicity by DP.     

A Critical Evaluation of Predicting Ocular lrritancy Potential from an in Vitro Cytotoxicity Assay

A Critical Evaluation of Predicting Ocular Irritancy Potentialfrom an in Vitro Cytotoxicity Assay. KENNAH, H. E., II, ALBULESCU,D., HIGNET, S., AND BARROW. C. S. (1989) Fundam Appl. Toxicol12,281-290. Numerous in vitro cytotoxicity assays have beenproposed as potential alternatives to the Draize eye irritancytest. The results reported, based upon the rank correlationof ocular irritancy with cytotoxicity, have been encouraging.However. direct calibration of in vivo to in vitro data utilizingseveral categories of chemicals has not been reported. Thisstudy evaluated the use of in vitro cytotoxicity data for predictingthe ocular irritancy potential of 24 chemicals(six surfactants,seven alcohols four ketones, four acetates and three aromatics).BALB/c 3T3 cells were grown overnight, then exposed for 30 minto at least four different concentrations of each chemical (expressedas volume percentage). Linear regression analysis of the logconcentration versus percentage of control growth was used tocalculate the concentration of toxicant that inhibited the normalgrowth rate by 50% (G150). The rank ordering of cytotoxicitybased upon the GI50s was surfactants > aromatics > alcohols> ketones or acetates. The larger molecular weight representativeof each senes (i.e., 2-ethyl-I-hexanol for alcohols) had lowerGI50 values than those of the lower molecular weight substances.The GI50 values were then directly calibrated against in vivoocular irritancy quantitated as percentage corneal swellingfollowing exposure of rabbits to the same test chemicals. Asignificant linear correlation between cytotoxicity and ocularirritancy was established only for surfactants and alcohols.For acetates, ketones, and aromatics there was little correlation.The overall poor correlation between cytotoxicity and ocularirritancy was attributed to differences in mechanisms of irritancy.The lack of correlation illustrates that in vitro cytotoxicitydata cannot be used to predict the ocular irritancy potentialof a broad spectrum of chemicals.     

In Vitro Cytotoxicity and Anti-tumor Properties of the Total Alkaloid Fraction of Unripe Fruits of Solanum pseudocapsicum

The total alkaloid fraction of the methanol extract of unripe fruits of Solanum pseudocapsicum was tested for in vitro cytotoxicity on HEp-2, RD and Vero cell lines and anti-tumor activity using DLA and HEp-2 cell lines. Cell viability and morphological changes were assessed. The total alkaloid fraction exhibited strong cytotoxic activity against all the cell lines tested. The CTC 50 (50 % of cytotoxicity inhibition) was found to be 1.65 µg/ml for the RD cell line, 6.32 µg/ml for the HEp-2 cell line and 12.01 µg/ml for the Vero cell line. In the clonogenic assay, no colony formation was observed even up to a concentration of 25 µg/ml. In the short term, antitumor, studies using DLA cells, the total alkaloid fraction was associated with 50 % viability in the concentration range of 6.25-12.5 µg/ml. In long term, anti-tumor activity using the HEp-2 cell line, no colony formation was observed up to a concentration of 20 µg/ml. Hence, there is a correlation between the results obtained in the cytotoxicity and antitumor studies carried out. Morphological observation by phase contrast microscopy revealed intense damage on all the cell lines. The total alkaloid fraction has the potential for further investigation in animal models.     

Chemical Characterization and in Vitro Cytotoxicity on Squamous Cell Carcinoma Cells of Carica Papaya Leaf Extracts

In traditional medicine, Carica papaya leaf has been used for a wide range of therapeutic applications including skin diseases and cancer. In this study, we investigated the in vitro cytotoxicity of aqueous and ethanolic extracts of Carica papaya leaves on the human oral squamous cell carcinoma SCC25 cell line in parallel with non-cancerous human keratinocyte HaCaT cells. Two out of four extracts showed a significantly selective effect towards the cancer cells and were found to contain high levels of phenolic and flavonoid compounds. The chromatographic and mass spectrometric profiles of the extracts obtained with Ultra High Performance Liquid Chromatography-Quadrupole Time of Flight-Mass Spectrometry were used to tentatively identify the bioactive compounds using comparative analysis. The principal compounds identified were flavonoids or flavonoid glycosides, particularly compounds from the kaempferol and quercetin families, of which several have previously been reported to possess anticancer activities. These results confirm that papaya leaf is a potential source of anticancer compounds and warrant further scientific investigation to validate the traditional use of papaya leaf to treat cancer.     

Kinetic Analysis on the Skin Disposition of Cytotoxicity as an Index of Skin Irritation Produced by Cetylpyridinium Chloride: Comparison of In Vitro Data using a Three-Dimensional Cultured Human Skin Model with In Vivo Results in Hairless Mice

PURPOSE: The aim of this study was to kinetically and dynamically analyze in vitro cytotoxicity as an index of skin irritation by use of a three-dimensional cultured human skin model and to compare the in vitro assay data with data from living animals. METHODS: A cationic surfactant, cetylpyridinium chloride (CPC), was selected as a model irritant. Living skin equivalent-high (LSE-high) and hairless mice were used for the in vitro and in vivo tests, respectively. Skin irritation dermatodynamics was evaluated by calorimetric thiazoyl blue (MTT) conversion assay both for in vitro and in vivo tests, whereas dermatokinetics of CPC in LSE-high and mouse skin were evaluated using HPLC. RESULTS: The time course of cell viability in the skin after application of CPC to intact skin was distinctly different from that of stratum-corneum-stripped skin in both LSE-high and hairless mice. Biphasic behavior characterized by two first-order rates with an inflection time point was observed in intact skin, whereas cell viability monoexponentially decreased immediately after CPC application in stripped skin. The time courses of cell viability in the skin and dermatodynamics were closely related to that of dermatokinetics of CPC. CONCLUSION: The present study demonstrates that the in vitro cytotoxic profile was similar to the in vivo cytotoxicity test and that dermatodynamics was related to dermatokinetics of CPC.     

MTT比色法与相对增殖度法对输液用包装材料的细胞毒性检测结果的比较

目的 比较四唑盐(MTT)比色法和相对增殖度法对4种输液用包装材料细胞毒性检查结果的相关性,评价2种方法在检测药包材体外细胞毒性作用上的优缺点。为建立快速有效的药包材检测新标准提供有力依据。方法 L-929细胞分别采用MTT法和细胞增殖度法对4种包装材料的浸提液进行细胞毒性研究,比较材料对细胞的毒性等级,算出细胞的相对增殖度,分析比较2种方法相关性。结果 在同样实验条件下,4种药品包材表现出对L-929细胞不同的细胞毒性。与细胞增殖度法比较,MTT法与细胞增殖法检测结果具有很好的相关性r=0.966(P<0.05)。结论 与相对增殖度法相比,MTT比色法由于实际检测所需的细胞量相对较少、试验步骤相对简便、检测周期短,因此具有一定的优越性,值得推荐作为细胞毒性检测方法。     

Effect of Ultrafilterable Platinum Concentration on Cisplatin and Carboplatin Cytotoxicity in Human Tumor and Bone Marrow Cells in Vitro

The importance of the ultrafilterable platinum (fPt) fraction of cisplatin (CDDP) and carboplatin (CBDCA) for cytotoxicity and myelotoxicity was studied in vitro. By incubating CDDP or CBDCA with fetal calf serum (PCS) various fractions of fPt were prepared and determined by atomic absorption spectroscopy. A relation of % fPt fraction and incubation time (h) of 87^e-1123t(r = –0.99) and 101^e-o.oo87t (r = –0.99) were determined for CDDP and CBDCA, respectively. Cytotoxicity in the human small cell lung carcinoma cell line GLC₄ and fPt fraction were closely related for CDDP (r = 0.99) and for CBDCA r = 0.97). However, at a similar fPt fraction the concentrations inhibiting cell survival by 50% (IC50) of CBDCA exceeded that of CDDP by a factor of 10-18 with 4 h exposure and a factor of 5 with continuous exposure. Tested in the range of peak concentrations in plasma of patients and at a clinically relevant fPt fraction of 10%, CDDP was not toxic for human bone marrow cells in the CFU-GM assay, whereas it was toxic at fPt fractions of 50% and 90%. However, CBDCA was myelotoxic at a (clinically relevant) fPt fraction of 50%, and also at 75% and 90%. The use of different fPt fractions, produced by the incubation method described in this study, permits the study of platinum drugs in vitro while approximating in vivo conditions might be used to evaluate myelotoxicity of new platinum drugs prospectively. For CDDP and CBDCA the fraction fPt determines cytotoxicity on tumor cells, and their different fPt fraction in patients account at least partly for their difference in myelotoxicity.     

Metoclopramide Inhibits the Cytotoxicity of Cisplatin and Enhances the Cytotoxicity of Epirubicin

Abstract: During cancer treatment, the use of antiemetics are often needed due to induction of nausea and vomiting by antineoplastic drugs. Some antiemetics have however been afflicted with cytotoxic effects during chemotherapy. The influence of the commonly used antiemetic metoclopramide on the cytotoxicity of epirubicin and cisplatin was tested on fibroblasts (V79) and lung cancer cells (P31) in vitro. The clonogenic survival of fibroblast and lung cancer cells were reduced when the cells were exposed to epirubicin or cisplatin. Metoclopramide (0.5 or 5 mg/l) enhanced epirubicin-induced toxicity to both fibroblast and lung cancer cells, but inhibited the cytotoxicity of cisplatin. The demonstrated effect of metoclopramide on cells in vitro and the fact that metoclopramide is used as a routine antiemetic during cancer treatment, may indicate that a possible clinical interaction with the antineoplastic action of cancer treatment drugs should be given attention.     

皂苷Rb1活性代谢产物NG701聚合物载药胶束的制备及其体外细胞毒性评价

目的:制备二醇类皂苷Rb1活性代谢产物(NG701)的聚合物载药胶束,并对其进行体外细胞毒性评价。方法:合成两亲性嵌段共聚物即聚乙二醇-聚天冬氨酸苄酯(PEG-PBLA)并借助红外光谱和核磁共振氢谱进行表征;采用透析法制备NG701/PEG-PBLA聚合物载药胶束,观察其形态和粒径分布,建立高效液相色谱法测定其含量并计算载药量和包封率。以Hela细胞为模型,采用MTT法评价聚合物载药胶束的细胞毒性作用,并与NG701原料药比较。结果:红外光谱和核磁共振氢谱证实形成了PEG-PBLA,所制备的聚合物载药胶束呈球形,粒径小于200nm,载药量最高达32.7,包封率最高为93.2。与原料药发挥细胞毒性作用的有效浓度100μg.mL-1比较,聚合物载药胶束为50μg.mL-(1P<0.05)。结论:以PEG-PBLA为载体制备的载药胶束聚合物,可有效提高NG701的细胞毒性作用。     

经导管瓣膜镍钛合金支架镍离子释放体外模拟及细胞毒性评价

目的：研究经导管瓣膜镍钛合金支架动态疲劳中镍离子释放及其潜在体外细胞毒性,为该类创新医疗器械产品的临床前安全性评价提供参考,为推动医疗器械创新发展提供技术支撑。方法：以经导管瓣膜镍钛合金支架为研究对象,利用脉动疲劳试验模拟支架临床植入10年的情况(4亿次疲劳测试),研究其动态疲劳过程中镍离子的释放情况,并利用细胞模型评估所释放镍离子的潜在毒性效应。结果：随着镍钛合金支架疲劳试验时间的增加,测试溶液中镍离子浓度增大,测试液中镍离子最大浓度为0.075 mg·L^-1。 利用不同浓度梯度的镍离子溶液对L929细胞进行细胞毒性试验,当离子浓度为小于等于6.25 mg·L^-1时, 无体外细胞毒性,当镍离子浓度大于等于12.5 mg·L^-11时,有潜在的细胞毒性。结论：经4亿次动态疲劳测试后(模拟临床使用10年)经导管瓣膜支架中镍离子的释放量低于安全限值,无体外细胞毒性。     

In Vitro Cytotoxicity of Mono-, Di-, and Trichioroacetate and Its Modulation by Hepatic Peroxisome Proliferation

Dichloroacetate (DCA) and trichloroacetate (TCA) are major by-productsof drinking water chlorination. Recent experiments have shownthat both of these compounds produce hepatic tumors in B6C3FImice. There was evidence that these effects may be associatedwith cytotoxic effects and/or peroxisomal proliferation. Therefore,in the present study the in vitro cytotoxicity of monochloroacetate(MCA), DCA, TCA and a metabolite, glycolate (GLY), was determinedin hepatocyte suspensions prepared from naive and clofibricacid-pretreated male Sprague-Dawley rats and B6C3F1 mice. Cytotoxicresponses, measured by release of lactic dehydrogenase and/ortrypan blue exclusion, were only observed with high concentrations(5.0 mM) of MCA and GLY in hepatocytes from naive animals (p=0.025and 0.008, respectively, Sprague-Dawley rat; p=0.033 and 0.001,respectively, B6C3F1 mouse). The cytotoxic responses to bothcompounds were observed much earlier and at much lower concentrationsin hepatocytes taken from mice and rats that had been pretreatedwith clofibric acid (p0.001, Sprague-Dawley rat and B6C3F1 mouse).DCA and TCA produced no evidence of cytotoxicity in hepatocytesfrom naive or clofibric acid-pretreated animals of either speciesat concentrations up to 5.0 mM. Increasing concentrations ofMCA and GLY resulted in dose-related depletion of intracellularreduced glutathione (GSH) that closely paralleled the cytotoxicresponses. Only GLY (0.25–5.0 mM) produced increased intracellularoxidized glutathione. Neither DCA nor TCA was found to altercellular GSH status in hepatocytes isolated from either Sprague-Dawleyrats or B6C3F1 mice. It was concluded from these in vitro observationsthat DCA and TCA are not highly cytotoxic to hepatocytes. Moreover,the rates of their conversion to MCA or GLY may be insufficientto induce cytotoxic effects in hepatocytes in vivo.     

A Rapid in Vitro Assay for Evaluation of Metabolism-Dependent Cytotoxicity of Antiepileptic Drugs on Isolated Human Lymphocytes

In vitro assessment of human lymphocyte viability by trypanblue dye exclusion in the presence of an external metabolizingsystem (microsomes plus NADPH) has been shown to be a usefulmethod in assessing predisposition to idiopathic toxicity inresponse to various anticonvulsant drugs. The trypan blue method,however, is labor intensive, is time consuming, is prone tohuman error, is not suitable for high-volume toxicity screening,and excludes autolysed cells. The objective of this study wasto develop a rapid, high-capacity, objective, and easy in vitrocytotoxicity method for the detection of metabolism-dependentcytotoxicity of a test chemical. The in vitro system uses anexternal metabolizing system (rabbit microsomes) in conjunctionwith isolated human lymphocytes as the target cells. Cellulartoxicity was determined by assessing plasma membrane integrityusing a membrane-impermeant fluorescent nucleic acid dye (YO-PRO-1)and a multiwell plate scanner for fluorescence. Using this system,cells incubated with either acetaminophen (1500 µg/ml),carbamazepine (62.5 µM), phenytoin (62.5 µm), orphenobarbital (62.5 µM) showed net increases in percentagecell death of 31 ± 5, 11 ± 4, 0 ± 3, and2 ± 3, respectively. A metabolism-dependent concentration-responsewas observed for valproic acid-induced cytotoxicity, which approacheda plateau at a concentration of 4000 µ/ml with a net percentagecell death of 31 ± 4. This technique resolves varioustechnical difficulties inherent in viability determinationsby the trypan blue exclusion method. The YO-PRO-1 method alsomay be useful in a clinical setting for the assessment of patientswith a genetically determined susceptibility to certain drugsand for identifying the responsible drug in patients with idiopathictoxicity undergoing multiple-drug therapy.     

Physicochemical,Pharmacokinetic and Cytotoxicity of the Compounds Isolated from an Endophyte Fusarium oxysporum: In Vitro and In Silico Approaches

The present study was intended to characterize the secondary metabolites of the endophyte Fusarium oxysporum isolated from the plant Aglaonema hookerianum Schott. And to investigate the cytotoxic and other pharmacological properties of the isolated compounds as part of the drug discovery and development process. Different chromatographic techniques were adopted to isolate the bioactive compounds that were identified by spectroscopic techniques. The cytotoxic properties of the compounds were assessed in the Vero cell line via the trypan blue method. Moreover, physicochemical, pharmacokinetic, bioactivity and toxicity profiles of the compounds were also investigated through in silico approaches. After careful spectral analysis, the isolated compounds were identified as 3β,5α-dihydroxy-ergosta-7,22-dien-6-one (1), 3β,5α,9α-trihydroxy-ergosta-7,22-dien-6-one (2), p-hydroxybenzaldehyde (3), 3-(R)-7-butyl-6,8-dihydroxy-3-pent-11-enylisochroman-1-one (4) and beauvericin (5). An in vitro study in the Vero cell line revealed that the presence of the compounds reduced the number of cells, as well as the percentage of viable cells, in most cases. An in silico cytotoxic analysis revealed that compounds 1, 2 and 5 might be explored as cytotoxic agents. Moreover, compounds 3 and 4 were found to be highly mutagenic. The present study suggested that further thorough investigations are necessary to use these molecules as leads for the cytotoxic drug development process.     

Cytotoxicity of corylifoliae fructus. II. Cytotoxicity of bakuchiol and the analogues

Bakuchiol is a major component of Corylifoliae Fructus (Psoralea corylifolia L.) and has been clarified to have cytotoxic activity. The chemical structure-cytotoxic activity relationship of bakuchiol was investigated by means of cytotoxic activity of synthesized analogues of bakuchiol and phenol. It was proved that an alkyl group was necessary for cytotoxic activity. But the double bonds in the unsaturated hydro-carbon group exerted but little influence on the cytotoxic activity. The cytotoxic activity of bakuchiol was the strongest as compared with that of the analogues examined.