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PubMed1981

Myelopoiesis in lymph nodes of mice. Enhancement effect by the injection of trypan blue or India ink after birth.

Myelopoiesis in lymph nodes was studied in newborn mice under normal conditions and after treatment with trypan blue and carbon particles. In the mouse lymph node, myelopoiesis normally takes place between the fourth and twelfth day after birth, then decreases while an intense proliferation of the lymphocytic population simultaneously takes place. Myelopoiesis manifests itself as clusters of cells, mainly of the neutrophilic line, located around post-capillary venules (PCV's) in the medulla. The injection of trypan blue or carbon particles (India ink) causes an enhancement of myelopoiesis and a delay in the development of the lymphocytic population. Myelopoiesis includes not only granulopoiesis but also the formation of erythroid clusters and occasional megakaryocytes. Three possibilities should be considered to explain the enhancement of myelopoiesis: 1) blockade of the reticuloendothelial system caused by overloading with particulate matter; 2) induction of humoral myelopoiesis-stimulating factors; and 3) stimulation of pluripotent stem cells through dye injection.

Animals↗
PubMed1989

Interaction of tetranectin with sulphated polysaccharides and trypan blue.

Tetranectin is a recently discovered plasminogen binding protein from human plasma, with a known primary structure. Its interactions with various sulphated polysaccharides and trypan blue has been investigated by crossed immunoelectrophoresis, immunosorbent assay (ELISA) and gel filtration experiments. Interaction of tetranectin with chondroitin sulphate A, B, and C, heparan sulphate and trypan blue could be demonstrated by crossed immunoelectrophoresis against monospecific rabbit anti-tetranectin. Interaction of tetranectin with fucoidan could be demonstrated by all three methods, and this interaction could be prevented by heparin and to a lesser degree by chondroitin sulphate A and C, but not by fucose. These findings, together with the recently published findings, that tetranectin is present in exocrine and endocrine glands as well as in secreting surface epithelia and mesenchyme, suggest that tetranectin, in conjunction with proteoglycans, may have a function in the packaging of granules or as a participant in exocytocic processes.

Animals↗
PubMed1975

The embryogenesis of trypan-blue induced spina bifida aperta and short tail in the rat.

The study of litters from 140 Wistar-derived rats injected with trypan blue during gestation leads to conclusion that a myelocele can result from faulty closure of the neural plate and that this is accompanied in many cases by blebs in the paraxial mesoderm. Haematomata usually underlie the open neural plate at an early stage and they form by extravasation of blood from the dorsal aortae into the blebs. Local ventral deflection of the notochord beneath a myelocele probably results from delayed separation of the notochord from the hindgut. Complete failure of separation and abnormality induced in the tail-bud could result in sacral agenesis and/or a short tail. All these malformations may result from varying severity of the action of the trypan blue at different developmental stages.

Age Factors↗
PubMed1975

[New muscle formation following transplantation of muscle treated with trypan blue].

The sources and mechanism of muscle formation de novo were investigated under the transplantation of skeletal muscle fibers treated in the 1% water solution of Trypan Blue for 48 hrs after Levander under skin and in omentum in rabbits and rats. In some cases the pieces of Trypan Blue stained muscles were placed in the diffusion chambers. Besides, 3H-thymidine autoradiography was used. It was established that after the above described treatment the muscles appeared morphologically as necrotized. They do not develop in the diffusion chambers in which cells do not penetrate. Under the transplantation of such muscles under skin and in omentum, they are phagocytized and disintegrate and in the close neighbourhood myoblasts arise which transform into muscle tubules and differentiated cross-striated muscle fibers. If prior to the transplantation the rats--recipients were labeled by 3H-thymidine, the muscle fibers formed de novo with the labeled nuclei, i. e. from the recipient cells in omentum where there are no muscles, apparently from polyblasts. Under the definite experimental conditions, myoblasts appear to arise from cells of non-myogenic origin by means of induction.

Animals↗
PubMedDate not supplied

Trypan blue staining of the anterior capsule: the one-drop technique.

Capsule staining is usually a two- or three-stage procedure in which trypan blue is injected under air, a viscoelastic device, viscoelastic mixed with dye, or compartmentalized viscoelastic and balanced salt solution. In these techniques, the consequent irrigation of the anterior chamber with balanced salt solution may lead to a less stable anterior chamber, a decrease of pupil diameter, and damage of the corneal endothelium. A modification of current injection techniques for staining the anterior capsule under a dispersive viscoelastic device with 1 drop of trypan blue by an ordinary 27-gauge anterior chamber cannula is described.

Capsulorhexis↗
PubMed1976

Combined trypan blue and formaldehyde-induced fluorescence in experimental skin freezing in the rat.

In order to study the effects of skin freezing in the rat on cutaneous vasculature and adrenergic nerves, a combination of trypan blue and formaldehyde-induced catecholamine fluorescence was used. It was shown that trypan blue clearly demonstrates both hyperaemic and ischaemic areas as well as changes in blood cessel permeability after experimental cold injury. Combination of the two fluorescence methods made it possible to correlate rough changes in blood vessel flow and permeability with changes in adrenergic nerves. Twentyfour hours after freezing of the skin, denervation of the most seriously damaged area was apparent. At the border of the cold injury, signs suggesting accumulation of actecholamines were observed in the subcutaneous nerves.

Animals↗
PubMed1999

Trypan blue capsule staining to visualize the capsulorhexis in cataract surgery.

A capsulorhexis may be difficult to perform in the absence of a red fundus reflex. Using 0.1 mL of trypan blue 0.1% to stain the anterior capsule in 30 patients with a mature cataract enabled us to visualize the capsulorhexis during phacoemulsification. No adverse reactions were observed up to 12 months after surgery. Trypan blue staining of the anterior capsule appears to be a safe technique to facilitate the performance of a capsulorhexis in the absence of a red fundus reflex.

Capsulorhexis↗
PubMed1992

Activation of mouse oocytes, fertilization and development of mouse embryos in vitro after staining with trypan blue or fluorescein diacetate.

The influence of vital staining with trypan blue or fluorescein diacetate on the fertilization of mouse oocytes and the developmental potential of mouse embryos was assessed. Neither stain induced spontaneous activation in mouse oocytes, nor did they impair the in vitro development and implantation of mouse zygotes, two-cell embryos, stressed morulae or blastocysts. However, fertilization and subsequent development of mouse oocytes have been shown to be reduced by vital staining.

Animals↗
PubMed1999

[Staining the lens capsule with trypan blue for visualizing capsulorhexis in surgery of mature cataracts].

BACKGROUND: A capsulorhexis may be difficult to perform in the absence of a red fundus reflex. PATIENTS AND METHODS: After application of 0.3 mL trypan blue 0.1%, a quick and homogeneous staining of the anterior lens capsule was obtained in 100 patients with a mature cataract, to visualize the capsulorhexis during a phacoemulsification procedure. RESULTS: No adverse reactions related to the dye were observed up to 18 months after surgery. CONCLUSION: Trypan blue staining of the anterior lens capsule may therefore be a safe technique to facilitate the performance of a capsulorhexis in the absence of a red fundus reflex.

Capsulorhexis↗
PubMed1987

Malformations in rat fetuses induced by trypan blue.

Malformations of fetuses obtained from Wistar rat dams treated with trypan blue during gestation were studied. Fetuses were examined on day 20 of gestation. One hundred and twenty-seven fetuses showed abnormalities of the external features, skeleton and internal organs, separately or in combination. External malformations were found in 108 fetuses. The most frequent external malformation was anomaly of tail. Spina bifida, club foot, exencephaly and anal atresia were also observed frequently. Skeletal malformations were detected in 48 fetuses. Deformity of vertebrae in the lumbar, sacral and/or caudal regions was found in 46 fetuses. Internal malformations were observed in 27 fetuses. Anomaly of heart and/or great vessels, hydrocephaly and micro- or anophthalmia were observed frequently. About 90% of the fetuses with skeletal malformations also showed some external malformations. In contrast, about 48% of the fetuses with internal malformations also had some external malformations. These results suggest that, for teratological study, internal examination is more important in detecting malformations of fetuses than skeletal examination.

Abnormalities, Drug-Induced↗
PubMed1983

Electrocardiographic studies of developing rodents exposed in utero to trypan blue.

Techniques, based on computer-assisted analyses of ensembled ECG waveforms, have been developed in this laboratory to serve as a screening procedure for potentially cardiotoxic substances. This procedure was tested on fetal and neonatal rats exposed in utero to two levels of trypan blue, a known cardiovascular teratogen. The offspring of the treated animals exhibited decreased viability, as well as significant levels of both cardiac and noncardiac malformations. Qualitative assessment of ECG irregularities demonstrated an increased number of abnormalities in the treated groups. Correlations were also obtained between observed morphological defects and ECG irregularities in the trypan blue treated groups.

Abnormalities, Drug-Induced↗
PubMed1990

Ultrarapid freezing and thawing of hamster oocytes. Morphologic parameters, trypan blue staining and sperm penetration assay for evaluating survival.

Nine hundred sixteen hamster oocytes were cryopreserved with the ultrarapid freezing method using five different cryoprotective solutions: 3 mol/L dimethylsulphoxide (DMSO) plus 0.25 mol/L sucrose, 3 mol/L DMSO, 3 mol/L propanediol plus 0.25 mol/L sucrose, 3 mol/L propanediol and 10% glycerol. One hundred eighty fresh oocytes served as controls. The viability of the oocytes was evaluated using morphologic parameters, Trypan blue staining and the sperm penetration assay. The viability rates based on morphologic parameters and Trypan blue staining were 82.3%, 65.0%, 51.4%, 33.0% and 0%, respectively, as compared to 100% in the controls. The sperm penetration rates were 27.0%, 0%, 9.8%, 0% and 0%, respectively, as compared to 94-98% in the controls. Our results indicate that among the various cryoprotective solutions used for ultrarapid freezing, 3 mol/L DMSO plus 0.25 mol/L sucrose gave the best results, with a viability rate of 82.3% and a sperm penetration rate of 27%.

Animals↗
PubMed1983

Ultrastructure of trypan blue induced ocular defects: I. Retina and lens.

The histological and cytological basis of trypan blue-induced ocular defects were studied using scanning and transmission electron microscopy. Microphthalmic and anophthalmic eyes of 16-day rat fetuses were utilized from dams exposed to a teratogenic dose of trypan blue. Retinal and lenticular anlagen were specifically examined for architectural and cellular changes. Nearly all severely abnormal eyes showed no evidence of retina development: Of 41 such eyes, only two retinal rudiments were observed. Those eyes with mild microphthalmia always demonstrated retinae although architectural changes were present. In every abnormal eye, some degree of lenticular morphogenesis was always present. Lenses were small, displaced in the eye field, and arrested at the lens vesicle stage. Lens cells were markedly undifferentiated and thus lacked most of the cytological features normally present at this developmental stage. Neither retinal nor lenticular rudiments were necrotic despite major architectural and cytological disturbances. The data offer three conclusions: First, the absence of necrosis suggests that trypan blue causes developmental arrest in this eye model; second, absence of retinae is most likely due to primary failure of optic vesicle development; third, lack of lens differentiation is attributed to absence of the retina, the primary lens inducer.

Abnormalities, Drug-Induced↗
PubMed1999

The evaluation of corneal graft tissue by the use of trypan blue.

BACKGROUND: The quality of the donor cornea, which basically depends on the condition of the endothelial sheet, is an important factor for the successful outcome of penetrating keratoplasty. PATIENTS AND METHODS: In this retrospective study, we present our experience in using trypan blue staining for the examination and evaluation of 498 consecutive corneal grafts from 251 donors, concerning their suitability for penetrating keratoplasty, during the decade 1986-1995. RESULTS: Forty-two grafts out of 498 (8.5%) were considered not suitable for surgery because of the great number of destroyed cells (cell density less than 1,500 cells/mm2) and the abnormal cell size and cell shape. Nine out of the 456 (1.97%) operated eyes exhibited primary donor failure and needed re-operation, probably because of boundary cell density of the graft (ranged in those cases from 1,600 to 1,800 cells/mm2). No infections of grafts were observed. CONCLUSION: Tissue evaluation of corneal grafts by the use of trypan blue staining is a simple, cost-effective, safe and accurate method that does not influence the vitality of the endothelial layer of the grafts.

Adolescent↗
PubMed2003

Contamination of trypan blue with Burkholderia cepacia in a cornea bank.

OBJECTIVE: To describe Burkholderia cepacia contamination of a cornea bank and the measures taken to identify and eliminate the source of infection. METHODS: Cultures were performed to assess the extent and source of contamination, and pulsed-field gel electrophoresis was used for molecular typing. RESULTS: Routine surveillance cultures identified 5 contaminated corneas during a 10-day period. Additional cultures showed that 28 of 88 samples were positive for this organism. Environmental investigation showed that an open bottle of trypan blue used to assess corneal morphology was contaminated with the epidemic strain. CONCLUSION: Trypan blue played a major role in this contamination of corneas. This episode shows that microbial contamination can affect transplanted corneas despite ongoing culture surveillance and suggests that new methods may be needed to avoid this risk.

Burkholderia Infections↗
PubMed1984

Trypan blue method for the identification of areas of damage to airway epithelium due to mechanical trauma.

A method has been developed for locating the regions of pulmonary epithelial damage due to mechanical trauma, based on the inability of the injured cells to exclude vital stains such as trypan blue. Scanning electron microscopy was used to confirm the sites, as well as the specific type of mechanical damage. The damaged area on rabbit tracheal epithelium due to gentle stroking was characterized by denudation of the ciliated epithelial cells, leaving only a layer of flat, non-ciliated cells. Repeated gentle stroking of the epithelial surface removed all the epithelial cells, exposing the underlying connective tissue layer. The trypan blue method provides a fast and sensitive means of locating damaged airway epithelium due to mechanical trauma. Because the dye is non-toxic, the repair process of the epithelium can also be studied, following the initial assessment of the extent of epithelial damage.

Animals↗
PubMed2004

Crystalline lens capsule staining with trypan blue.

A 1-step method for staining the anterior lens capsule with trypan blue is described. The dye is instilled via a paracentesis port at the start of cataract extraction. As aqueous humor is allowed to exit the anterior chamber (AC), which consequently shallows, the resulting pupil block confines the dye to the AC. An ophthalmic viscosurgical device (OVD) is used to flush dye-stained aqueous from the AC, circumventing the need for AC washout. Although the OVD may be tinged with dye, this does not impede performing capsulorhexis. This method does not add to the surgical time, requires no additional instruments or materials, and is safe.

Capsulorhexis↗
PubMed2003

In vitro assessment of the cytotoxicity of nisin, pediocin, and selected colicins on simian virus 40-transfected human colon and Vero monkey kidney cells with trypan blue staining viability assays.

Gram-positive bacterial bacteriocins (nisin and pediocin) and gram-negative bacterial bacteriocins (colicins [Col] E1, E3, E6, E7, and K) were evaluated for cytotoxicity against cultured simian virus 40-transfected human colon (SV40-HC) and Vero monkey kidney (Vero) cells. Bacteriocin-treated cells were assessed for viability by trypan blue staining. Monolayers of SV40-HC and Vero cells were cultured in tissue culture plates (35 degrees C, 10% CO2 in humidified air) with the use of Dulbecco's modified Eagle's medium supplemented with 10% (vol/vol) calf serum. Actively growing cells in the log phase (ca. 10(4) cells per ml) were treated with individual partially purified bacteriocin preparations at 170, 350, and 700 activity units per ml. Duplicate culture plates for each bacteriocin treatment and untreated controls were withdrawn after 16, 32, and 48 h of incubation. Cells were dissociated with trypsin and treated with trypan blue and were then counted in a hemocytometer with the use of a phase-contrast microscope. Viability assays indicated dose-dependent toxicity for some bacteriocins. Nisin, pediocin, and Col E6 were the most cytotoxic bacteriocins; SV40-HC cells demonstrated greater sensitivity than Vero cells did. Some bacteriocins can be toxic to mammalian cells; therefore, bacteriocins intended for use as biopreservatives must be evaluated for toxicity to mammalian cells and for other toxicities. Col E1, Col E3, Col E7, and Col K demonstrated little toxicity at the activities tested, indicating that they are safe and thus have potential for use as food biopreservatives.

Animals↗