Toluidine blue staining for cell and tissue biology applications

Vidal, Benedicto de Campos; Mello, Maria Luiza S.

doi:10.1016/j.acthis.2018.11.005

Cited by 30 publications

(28 citation statements)

References 76 publications

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“…, and COO − radicals present in GAGs and nucleic acids. 23,24 In the control group, we observed cells with metachromatic cytoplasm, slightly orthochromatic nuclei, and metachromatic nucleoli. On the scaffolds studied (NA, A150, and A300), we often found elongated cells and slightly more intense metachromasia.…”

Section: Cytochemical Analysismentioning

confidence: 77%

Morphological and biochemical analysis of cells cultured on fibrous poly(ε‐caprolactone) scaffolds with different degrees of fiber alignment produced by solution blow spinning

et al. 2023

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BackgroundBioresorbable materials are compounds that decompose in physiological mediums both in vitro and in vivo and are used as an alternative to temporary implants in injured tissues. The aim of this study was to analyze the morphology and cytochemistry of cells grown on fibrous poly(ε‐caprolactone) (PCL) scaffolds and to measure cell metabolism parameters by biochemical analysis of the conditioned culture medium from cells grown on the scaffolds.MethodsFibrous PCL scaffolds were used under the following conditions: unaligned fibers (NA), fibers aligned at 150 rpm (A150), and fibers aligned at 300 rpm (A300). Vero cells were cultured on these scaffolds for 24 h, 48 h, and 72 h. Samples were analyzed by SEM, MicroCT, cytochemistry, and culture medium biochemistry.ResultsThe results of the cytochemical analysis showed cells were confluent and well spread on the culture plate, while cells grown on the polymeric scaffold, exhibited an elongated morphology. In the biochemical analyses, no significant differences were observed in the expression of alkaline phosphatase or in the levels of cholesterol or total protein in the culture medium. The different materials do not seem to promote changes in the expression or metabolism of these molecules. Only glucose was markedly reduced in the culture medium of cells grown on either aligned or unaligned scaffolds for 48 h or 72 h. This finding indicates the intense energy requirements of cells grown on these scaffolds.ConclusionPCL fibers showed a great capacity to support cell growth. These data reinforce the interpretation that cells grow satisfactorily on PCL scaffolds.

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Section: Cytochemical Analysismentioning

confidence: 77%

Morphological and biochemical analysis of cells cultured on fibrous poly(ε‐caprolactone) scaffolds with different degrees of fiber alignment produced by solution blow spinning

et al. 2023

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“…To do so, 3 × 10 6 cells/ml ( n = 4) suspended in low glucose DMEM medium with 10% FBS were inoculated on the materials and the plate was cultivated for 24 h at 37 °C. The cells were then fixed in formaldehyde 10% for 24 h and stained with toluidine blue (TB) at pH 4.0 to detect the presence of nucleic acids and glycosaminoglycan acids [ 16 , 17 ]. The images were obtained with a Leica inverted microscope (model DM IL, Leica Microsystems, Wetzlar, Germany) with 10× and 20× objective lens.…”

Section: Methodsmentioning

confidence: 99%

Osteogenic differentiation of rat bone mesenchymal stem cells cultured on poly (hydroxybutyrate-co-hydroxyvalerate), poly (ε-caprolactone) scaffolds

Rodrigues

Batista

Malmonge

et al. 2021

J Mater Sci: Mater Med

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Bioresorbable biomaterials can fill bone defects and act as temporary scaffold to recruit MSCs to stimulate their differentiation. Among the different bioresorbable polymers studied, this work focuses on poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) and poly(ε-caprolactone) (PCL). Were prepared blends of PHBV and PCL to obtain PHBV based biomaterials with good tenacity, important for bone tissue repair, associated with biocompatible properties of PCL. This study assesses the viability of Vero cells on scaffolds of PHBV, PCL, and their blends and the osteogenic differentiation of mesenchymal stem cells (MSCs). Materials were characterized in surface morphology, DSC and Impact Strength (IS). Vero cells and MSCs were assessed by MTT assay, cytochemical and SEM analysis. MSC osteogenic differentiation was evaluated through alizarin red staining and ALP activity. We found some roughness onto surface materials. DSC showed that the blends presented two distinct melting peaks, characteristic of immiscible blends. IS test confirmed that PHBV-PCL blends is an alternative for increase the tenacity of PHBV. MTT assay showed cells with high metabolic activities on extract toxicity test, but with low activity in the direct contact test. SEM analysis showed spreading cells with irregular and flattened morphology on different substrates. Cytochemical study revealed that MSCs maintained their morphology, although in smaller number for MSCs. The development of nodules of mineralized organic matrix in MSC cultures was identified by alizarin red staining and osteogenic differentiation was confirmed by the quantification of ALP activity. Thus, our scaffolds did not interfere on viability of Vero cells or the osteogenic differentiation of MSCs.

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“…Moreover, toluidine blue is a known antidote used in patients with methemoglobinemia, which indicates the safety of intravenous and injection administration of this compound [ 27 ]. However, the histological staining of tissue sections remains the main field of its application [ 28 ].…”

Section: Introductionmentioning

confidence: 99%

Toluidine blue for the detection of sentinel lymph nodes in patients with thyroid cancer

Moskalenko¹,

Kurochkin²,

Vynnychenko³

et al. 2022

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Introduction Thyroid cancer (TC) demonstrates steady growth in incidence worldwide and remains an urgent problem in oncology. The detection of sentinel lymph nodes (SLN) with a selective dye and further histological examination in selecting the proper (personalized) surgical strategy and the volume of surgical intervention for clinically undetermined regional lymph nodes. The purpose of the study is to evaluate the effectiveness and safety of intraoperative detection of SLN with a 1% toluidine blue aqueous solution. Material and methods The significant tasks are to identify the pattern of TC metastases to cervical lymph nodes, to establish the prevalence of “skip” metastases, to compare the frequency of complications after total thyroidectomy and central neck dissection and lateral neck dissection with total thyroidectomy and central neck dissection, and to determine the feasibility of the application of lateral neck dissections in patients with papillary and follicular TC without metastases to regional lymph nodes (according to physical examination and ultrasound). Results According to our data the SLN identification rate was 97.6%. Sensitivity, specificity, positive predictive value, negative predictive value, and frequency of false negative and false positive results was 89.2, 94.6, 88.03, 95.16, 10.8, and 5.4%, respectively. The most common metastasis was in the central neck compartment (83.7%). Skip metastases were determined in 4.9% of patients. Conclusions The low prevalence of “skip” metastases and a significant risk of postoperative complications (wound exudation, lymphorrhagia, inflammation, hypoparathyroidism, paresis of the vocal cords) support the idea that lateral neck dissection is appropriate only in cases of confirmed metastases by physical examination and/or ultrasound at the preoperative stage.

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Toluidine blue staining for cell and tissue biology applications

Cited by 30 publications

References 76 publications

Morphological and biochemical analysis of cells cultured on fibrous poly(ε‐caprolactone) scaffolds with different degrees of fiber alignment produced by solution blow spinning

Morphological and biochemical analysis of cells cultured on fibrous poly(ε‐caprolactone) scaffolds with different degrees of fiber alignment produced by solution blow spinning

Osteogenic differentiation of rat bone mesenchymal stem cells cultured on poly (hydroxybutyrate-co-hydroxyvalerate), poly (ε-caprolactone) scaffolds

Toluidine blue for the detection of sentinel lymph nodes in patients with thyroid cancer

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