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PubMedDate not supplied

On the prenatal noxious effects of trypan blue and of a related azo dye.

Since 1948 trypan blue has been a well-known and extensively used experimental teratogen, belonging to the group of azo dyes. Chemically, trypan blue consists of a biphenyl molecule (0-tolidine or benzidine) combined by means of azo linkages with two molecules of a substituted naphthalene. Between 1987-89 the effect of the replacement of the biphenyl molecule by a molecule of p,p'-diaminobenzanilide upon the prenatal noxious action of trypan blue has been controlled. Investigations were carried out on three species: chick embryos, albino rats and albino mice. In the species used, the replacement annihilates the teratogenic properties of the dye, with the persistence of some embryotoxic effects. On the other hand, the control of o-tolidine and of p,p'-diaminobenzanilide revealed that no one had teratogenic properties (only some embryotoxic effect, more marked in the case of o-tolidine). It results that the teratogenic action of trypan blue cannot be attributed to the o-tolidine molecule proper but to an effect which results (in a for the moment unknown manner) from its combination with the other parts of the dye molecule.

Abnormalities, Drug-Induced↗
PubMed1981

Trypan blue inhibition of complement receptor function on various cells.

Trypan blue has previously been shown to inhibit complement-mediated phagocytosis by interaction with the C3 receptor but not with the Fc receptor. In studies reported here, trypan blue inhibited EAC3 rosette formation with human polymorphonuclear leucocytes (PMN) and mononuclear cells, rabbit alveolar macrophages (AM) and peritoneal PMN, and guinea-pig spleen cells. Trypan blue also inhibited C3-mediated bacterial adherence to the same receptor-bearing cells and to human glomerular cells. EAC3bi rosette formation was also inhibited, but EAC3d rosettes were not detected in our assay system. Although the precise molecular nature of C3b fragments deposited on antibody-sensitized erythrocytes has not been fully determined, trypan blue probably inhibits all C3 receptors from different species and may prove useful in vivo and in vitro for the definition of C3-receptor function in various aspects of the immune response.

Animals↗
PubMed2004

[Experimental study on the penetrability of trypan blue to the rat prostate].

OBJECTIVE: To understand the penetrability of trypan blue to the normal prostate as well as to the inflammatory prostate and the prostate with benign hyperplasia in rats. METHODS: Sixty SD male rats were randomized into 4 groups: NP (normal prostate) group (n = 15), BP (bacterial prostatitis) group (n = 15), BPH (benign prostatic hyperplasia) group (n = 15), and BPH-BP (benign prostatic hyperplasia with bacterial prostatitis) group (n = 15). Five rats were taken from each group as non-staining controls (NC, n = 5 x 4) and the other 10 were injected by tail intravenation with 1% trypan blue and then the prostates were isolated from the rats killed by anaesthesia after 2 hours. The color of the prostates and other tissues of the animals were observed and the contents of the trypan blue in the tissues of the prostates were determined separately by colorimetry. RESULTS: Apart from the tissues of brains and spinal cord the surface and the inner tissues of the prostates with NP, BP, BPH and BPH-BP from the rats injected with the dye were also dyed blue similar to that of the muscles, livers, intestines and others. The content of the trypan blue in the tissues of the prostates with NP, BP and BPH-BP was obviously higher than those with NP and BPH. CONCLUSION: The penetrability of trypan blue with properties of ionization and larger molecular weight is high in either the normal prostate or the prostate with BP, BPH and BPH-BP, and much higher in inflammatory prostate than in the normal prostate and the prostate with BPH.

Animals↗
PubMed1952

Studies on the generalized Shwartzman reaction. II. The production of bilateral cortical necrosis of the kidneys by a single injection of bacterial toxin in rabbits previously treated with thorotrast or trypan blue.

Intravenous injection of thorotrast or trypan blue rendered rabbits susceptible to the production of bilateral cortical necrosis of the kidneys by a single intravenous injection of small amounts of meningococcal or Serratia marcescens toxin. This reaction was not produced when thorotrast or trypan blue were injected after toxin had been given. A single intradermal injection of toxin produced hemorrhagic skin lesions resembling the local Shwartzman reaction in rabbits given thorotrast 6 hours previously. These animals also developed bilateral cortical necrosis of the kidneys. When the order of injection was reversed, and thorotrast given after toxin, neither skin nor kidney lesions occurred. The skin and kidney lesions in thorotrast-treated rabbits were, like the local and generalized Shwartzman reactions, completely prevented by treatment with nitrogen mustard, in doses sufficient to produce polymorphonuclear leukopenia. The significance of these reactions, and their relationship to the previously described response to toxin in cortisone-treated rabbits, are discussed.

Animals↗
PubMed2002

Trypan blue staining of epiretinal membranes in proliferative vitreoretinopathy.

OBJECTIVE: To determine whether trypan blue staining facilitates epiretinal membrane (ERM) removal in proliferative vitreoretinopathy. METHODS: In 10 patients undergoing vitrectomy for proliferative vitreoretinopathy, ERM peeling was performed without staining the tissue, until no additional ERMs were clearly visible. Then, after a fluid-air exchange, 0.06% trypan blue solution was applied onto the retinal surface. After 1 minute, all excess dye was removed and, after an air-fluid exchange, ERM peeling was completed. Excised ERM specimens were analyzed by transmission electron microscopy. MAIN OUTCOME MEASURES: For each patient, the efficacy of trypan blue staining of ERMs during surgery was scored. RESULTS: In all patients, intraoperative staining of ERMs with trypan blue was found to be a useful adjunct, since the dye consistently improved direct visualization and delineation of ERMs and facilitated ERM removal. A clear contrast was created between the stained ERMs and the nonstaining, underlying retina. Electron microscopy showed that only ERM tissue was removed. No adverse reactions related to the use of the dye were observed up to 3 months after surgery. CONCLUSIONS: Trypan blue may be an important new tool in the surgical management of proliferative vitreoretinopathy, since it may allow a more complete and safer ERM removal.

Aged↗
PubMed1989

[Trypan blue staining capacities of the culture cells (2)--The mechanism of staining].

The cultured cells dispersed by rubber policeman were stained with trypan blue, but these cells were viable at the meaning of keeping viral infective center forming capacity and at the meaning that trypan blue stained cells were transferred to unstained ones by trypsin treatment. This phenomenon may be important not to miscount numbers of the viable cells in cultured cells. Phagocytosis (endocytosis) is well known as a reaction inhibited by NaF or monoiodoacetate, but these metabolic inhibitors did not inhibit trypan blue staining of the cells dispersed both by rubber policeman and 0.05% trypsin treatment. The cells suspended in higher osmotic pressure media were more stainable with trypan blue than lower ones. Addition of K+ to the reaction mixture was not effective on trypan blue staining. These results suggest that "alive" cells dispersed by rubber policeman are stained with trypan blue not to utilize phagocytosis (endocytosis) or Na+-K+ pump, whose reactions couple with energy production. Trypan blue staining capacity of the cultured cells may be depended on pore sizes of protein channels (for example, fixed or gated transport protein channel) in lipid-bilayers of cell membranes. It is concluded that the cells dispersed by trypsin treatment (spherical shape in microscopical observation) and spherical cells in rubber policeman dispersing ones are unstained with trypan blue to contain smaller pore size channels in the cell membranes, but the rest, amoeba-like form cells dispersed by rubber policeman and dead cells are stained to contain larger ones.

Cell Survival↗
PubMed2005

Trypan blue identifies antimetabolite treatment area in trabeculectomy.

AIM: Colourless solutions of mitomycin C (MMC) and 5-fluorouracil (5-FU) are widely used during trabeculectomy to inhibit postoperative scarring. The poor visibility of these agents on the eye has several drawbacks including the inability to accurately assess the area of treatment. This study examined the utility of using trypan blue dye to colour antimetabolites used during trabeculectomy and the effect of trypan blue on antimetabolite cytotoxicity in vitro. METHODS: For in vitro experiments, MMC (0.4 mg/ml) and 5-FU (25 mg/ml) were reconstituted with or without trypan blue. A lactate dehydrogenase release assay was used to measure drug induced cell death and viable cell number 7 days after treatment. For clinical assessment, trypan blue 0.1% was added to MMC and 5-FU to final concentrations of between 0.01% and 0.05%. The mixture was applied to Tenon's capsule and sclera via pre-wet or into dry 5x8 mm sponges (MMC and 5-FU) for 3 minutes or by direct subconjunctival injection after completion of surgery (5-FU). Twenty two consecutive patients undergoing trabeculectomy either with or without trypan blue were followed for 2 years postoperatively. RESULTS: The addition of 0.05% trypan blue to MMC or 5-FU did not alter MMC induced cell death or the number of viable fibroblast in vitro. In vivo, trypan blue clearly delineated the antimetabolite treatment area and facilitated control of excess antimetabolite at the wound margins as well as sponge removal. With direct subconjunctival injection, total staining area varied for a given volume with location of the needle tip. Any leakage from the injection site could be easily seen. No adverse effects attributable to trypan blue were found in 2 years of follow up. CONCLUSIONS: Trypan blue permits delineation of antimetabolite/tissue interactions without affecting cytoxicity for the assays investigated. Trypan blue can be used to visualise antimetabolite soaked sponges, estimate treatment area, and show areas of unintended tissue contact during trabeculectomy. The addition of trypan blue to antimetabolites has potential benefits in clinical, research, and teaching aspects of ocular surgery and therapy.

Aged↗
PubMed2003

Epiretinal membrane surgery assisted by trypan blue.

PURPOSE: To evaluate patient outcome following epiretinal membrane surgery using trypan blue to facilitate visualization and delamination. DESIGN: A retrospective noncomparative review of 23 patients. METHOD: Patients underwent a three-port pars plana vitrectomy with delamination using trypan blue 0.06%. RESULTS: The median preoperative visual acuity was 20/100. The median postoperative visual acuity was 20/60. Seventeen patients (74%) improved their visual acuity by at least 2 chart lines. No adverse reaction related to trypan blue was observed up to 1 year postoperatively. CONCLUSIONS: Trypan blue staining of the epiretinal membrane facilitated visualization and delamination without any signs of toxicity.

Aged↗
PubMed2004

Trypan blue induces apoptosis in human retinal pigment epithelial cells.

PURPOSE: To examine whether trypan blue dye induces apoptosis in human retinal pigment epithelium cells. DESIGN: Laboratory investigation. METHODS: Pure cultures of human retinal pigment epithelium cells were isolated. The cells were incubated with different concentrations of trypan blue (0.5%, 0.10%, and 0.05%) for either 5 or 30 minutes. The rate of retinal pigment epithelium cell apoptosis was assessed with Annexin V-PE staining and flow cytometry. RESULTS: Trypan blue induced a statistically significant amount of apoptosis in retinal pigment epithelium cells at all the concentrations (0.5%, 0.10%, and 0.05%) (P <.05). The increase in incubation time (from 5 to 30 minutes) led to an increase in the number of apoptotic retinal pigment epithelium cells. CONCLUSION: The incubation of retinal pigment epithelium cells with trypan blue increased the number of apoptotic retinal pigment epithelium cells in vitro. Our results suggest that decisions regarding the intravitreal application of trypan blue dye need to be made with caution.

Annexin A5↗
PubMed1996

Effect of trypan blue on the action of acetylcholine, histamine and salbutamol in the isolated guinea-pig ileum.

It has been reported that trypan blue, a diazo dye with polyamphipatic structure, can inhibit the coupling of receptors to G-proteins. This inhibition of G-protein coupling has been investigated in isolated guinea-pig ileum. It was found that trypan blue could elicit a slight but dose-dependent contractile response in isolated guinea-pig ileum (4.5% of maximum contractile response induced by acetylcholine). While trypan blue potentiated the effect of histamine and shifted its dose-response curve to the left, it did not affect the contractile effects of acetylcholine. Furthermore, the relaxation which has been induced by salbutamol, a beta 2 agonist, was inhibited by trypan blue. It is concluded that trypan blue, as shown in biochemical studies, act selectively and can uncouple Gs-protein from beta 1 receptors. However, the effect of trypan blue on the whole tissue preparation depends on the type of G-protein involved and post G-protein processes which are stimulated after receptor activation. Trypan blue and similar agents could provide useful tools for further investigations of the mechanism of receptor-G protein coupling in the whole tissue preparation.

Acetylcholine↗
PubMed1998

Effects of trypan blue on the action of adrenergic agonists in the guinea-pig isolated atrium.

It has been reported that trypan blue, a diazo dye with polyamphipathic structure, can inhibit the coupling of receptors to G-proteins. The present study was carried out to investigate the effect of trypan blue on the actions of adrenoceptor agonists in the guinea-pig atrium. Trypan blue (10 and 100 microM) antagonized the positive inotropic effects of isoprenaline and dobutamine by shifting their concentration-response curves to the right. With the selective beta 2-adrenoceptor agonist, salbutamol, there was a reduction of response in the presence of trypan blue. Therefore, we concluded that trypan blue diminish the response to beta-adrenoceptor agonists possibly via decoupling receptors from Gs. Trypan blue and similar agents, due to their unique mode of action, can be used as tools for the investigation of the mechanism of receptor-G protein coupling in the whole tissue preparation.

Adrenergic Agonists↗
PubMed2004

[Results of surgery on white cataract using trypan blue capsule staining to visualize capsulorrhexis].

PURPOSE: To evaluate surgery on white cataracts using trypan blue capsule staining. METHODS: A retrospective study comprised 64 eyes of 60 patients with white cataract that had surgery with trypan blue capsule staining. The average age was 62.4 years and progress observation periods were 5.6 months. The rate of successful continuous curvilinear capsulorrhexis(CCC), complications, visual acuity, intraocular pressure(IOP), flare value, and corneal endothelial cell loss were studied. RESULTS: The CCC was completed uneventfully in 93.8% eyes. Posterior capsule rupture occurred in 2 eyes, and early perforation in 1 eye. Accidental vitreous staining and endothelial staining with trypan blue were observed in 1 eye each. There were no postoperative complications associated with trypan blue. Forty-five eyes had a best corrected visual acuity of 0.8 or better at the last visit. Twelve eyes had some ocular pathology resulting in visual loss, and a reliable visual acuity test could not be performed in 6 eyes. The mean postoperative IOP was within the normal range. The mean postoperative flare returned to within normal range 1 month after surgery. The mean corneal endothelial loss was 13.7%, and that of eyes with nucleus of grade 2 or softer was only 2.9%. CONCLUSIONS: Cataract surgery using trypan blue was safe and effective in managing white cataracts.

Adolescent↗
PubMed1977

Binding and uptake of trypan blue by developing oocytes of Locusta migratoria migratorioides.

Resorbing oocytes are heavily stained by trypan blue injected into the haemolymph; this serves as a basis for a quick and convenient method for measuring the degree of resorption. Oocytes in the beginning of their development are most susceptible towards resorptive tendencies. The uptake of trypan blue by normally developing oocytes is proportional to the oocyte surface. From 'double-marker' experiments, in which trypan blue is injected into the haemolymph together with [3H]inulin (which does not bind to the oocyte membrane) it is estimated that the contribution of binding in the interiorization of trypan blue is in the order of 80%, under the conditions given. In vitro incubations show the interaction of trypan blue with the membrane to be electrostatic in nature.

Animals↗
PubMed1979

Suppression of cell-mediated tumor cell lysis and complement-induced cytotoxicity by trypan blue.

Different forms of cell-mediated cytotoxicity were suppressed in the presence of trypan blue. The systems affected included lysis of antibody-coated tumor cells by normal and C. parvum-stimulated mouse peritoneal cells and lysis of allogeneic targets by immune effector cells. The inhibition, measured in a 4-hr 51Cr release assay, was reversible and did not occur in the presence of 30% fetal calf serum or albumin. Binding between effector and target cells through Fc receptors was not affected, and lysis of allogeneic cells was inhibited at the lytic step rather than at the binding step. In contrast, lysis of sensitized erythrocytes was not inhibited by trypan blue, suggesting that lysis of these targets may not involve the steps required in tumor cell lysis. Trypan blue blocked the function of antibody before binding to target cells and also suppressed complement-induced cytolysis. Most individual complement components were susceptible to the inhibitory action of trypan blue. These results reveal an affinity of trypan blue for proteins in general that may be responsible for many of its biologic actions.

Animals↗
PubMed2005

Toxic effects of indocyanine green, infracyanine green, and trypan blue on the human retinal pigmented epithelium.

BACKGROUND: Indocyanine green, infracyanine green, and trypan blue are frequently used as aids to visualize structures removed during vitreoretinal surgery. But they may have toxic effects on the retina. We therefore compared the acute and chronic toxicities of these stains on cultured human retinal pigmented epithelial (RPE) cells using clinically relevant concentrations and an identical experimental setup for each agent. METHODS: Monolayers of RPE cells were incubated with various concentrations of indocyanine green, infracyanine green (each at 0.005%, 0.05%, and 0.5%) or trypan blue (0.05%, 0.06%, 0.1%, 0.15%, and 0.5%) for 5 min (acute exposure) or 6 days (chronic exposure). Using the propidium iodide assay, acute cytotoxicity was monitored at 15-min intervals for up to 3 h. Chronic cytotoxicity was assessed by monitoring cell calcein esterase activity, cell proliferation, and cell morphology (viability) after 6 days of exposure. RESULTS: Indocyanine and infracyanine green induced acute and chronic toxicities at a concentration above 0.05%. Trypan blue evoked no acute toxicity, but it was chronically cytotoxic at all tested concentrations. CONCLUSIONS: Despite thorough rinsing after application, significant amounts of the not sufficiently water soluble indocyanine and infracyanine green are retained after surgery by the eye. Trypan blue, being more water-soluble than ICG, is probably retained to the least degree. This circumstance is fortunate given that trypan blue exhibits a chronic cytotoxicity comparable to ICG at all clinically relevant concentrations. During vitrectomy, surgeons should aim to expose retinal tissue to only low concentrations of these stains and for as short a period as possible.

Cell Proliferation↗
PubMed2003

Trypan blue staining in vitreoretinal surgery.

PURPOSE: To evaluate the efficacy of trypan blue for staining the internal limiting membrane (ILM) and epiretinal membranes (ERM) in vitreoretinal surgery. DESIGN: Prospective noncomparative case series. PARTICIPANTS: Fifty eyes of 50 patients with macular pucker (n = 22), macular hole (n = 18), or a combination (n = 2), proliferative vitreoretinopathy (n = 5), or diabetic retinopathy (n = 3). METHODS: Trypan blue 0.2% was used to stain the ILM or ERM during vitreoretinal surgery. MAIN OUTCOME MEASURES: The intraoperative visibility of the membranes was scored as poor, moderate, good, or excellent. RESULTS: The application of trypan blue onto the ILM or the ERM resulted in a useful bluish staining, facilitating the identification, delineation, and removal of the membranes in all surgeries. No residual staining or adverse effects related to the dye were observed. CONCLUSIONS: Trypan blue stains both ILM and ERM and might be an useful tool in vitreoretinal surgery.

Adult↗
PubMed1977

Permeability of inner and outer layers of rat and rabbit aortic wall. Two new microscopic test with trypan blue.

Two new permeability tests are described for use with intravenously injected trypan blue. One depends on the demonstration of trypan blue by its specific red fluorescence in green light at 570 nm, while the other is a surface microscopy technique at low magnification, using illumination from a fibre-optics light source. The routes of entry of the trypan blue-albumin complex into the rat and rabbit aorta appear to be from both the inner and outer surfaces. Over the period 1/4-24 h after injection of the dye, more entered from the outer surface than the inner surface in the rat aorta and rather more in the rabbit thoracic aorta. The arch of the rabbit aorta showed in general rather greater entry from the inner surface. Trypan blue that has entered the aortic wall is partly taken up by the elastic lamellae. Elastic competes successfully for the dye and captures it from the trypan blue-albumin complex; this uptake is blocked by deamination with nitrous acid.

Animals↗