Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 458
Filter
1.
Einstein (Säo Paulo) ; 20: eAO6450, 2022. tab, graf
Article in English | LILACS | ID: biblio-1364794

ABSTRACT

ABSTRACT Objective To understand the feasibility of FGFR3 tests in the Brazilian public health context, and to sample the mutational burden of this receptor in high-grade muscle invasive bladder cancer. Methods A total of 31 patients with high-grade muscle-invasive bladder cancer were included in the present study. Either transurethral resection of bladder tumor or radical cystectomy specimens were analyzed. Formalin-fixed paraffin-embedded tissue blocks were sectioned, hematoxylin and eosin stained, and histologic sections were reviewed. Total RNA was extracted using the RNeasy DSP formalin-fixed paraffin-embedded kit. Qualitative results were displayed in Rotor-Gene AssayManager software. Results Six patients were excluded. From the samples analyzed, four (16.7%) were considered inadequate and could not have their RNA extracted. Two patients presented FGFR3 mutations, accounting for 9.5% of material available for adequate analysis. The two mutations detected included a Y373C mutation in a male patient and a S249C mutation in a female patient. Conclusion FGFR3 mutations could be analyzed in 84% of our cohort and occurred in 9.5% of patients with high-grade muscle invasive bladder cancer in this Brazilian population. FGFR3 gene mutations are targets for therapeutic drugs in muscle-invasive bladder cancer. For this reason, know the frequency of these mutations can have a significant impact on public health policies and costs provisioning.


Subject(s)
Urinary Bladder Neoplasms/genetics , Carcinoma, Transitional Cell/genetics , Carcinoma, Transitional Cell/metabolism , Carcinoma, Transitional Cell/pathology , Receptor, Fibroblast Growth Factor, Type 3/genetics , Receptor, Fibroblast Growth Factor, Type 3/metabolism , Brazil , RNA , Prevalence , Eosine Yellowish-(YS) , Hematoxylin , Muscles/metabolism , Muscles/pathology , Mutation
2.
Rev. bras. ortop ; 55(6): 778-782, Nov.-Dec. 2020. graf
Article in English | LILACS | ID: biblio-1156183

ABSTRACT

Abstract Objectives The present paper aims to evaluate and compare the histological features of fresh and frozen menisci stored in a tissue bank for 1 month and for 5 years. Methods The meniscal grafts were subjected to a histological study. A total of 10 menisci were evaluated; 2 were frozen for 5 years, 4 were frozen for 1 month, and 4 were fresh, recently harvested specimens. Histological properties were evaluated in sections stained with hematoxylin and eosin and Masson trichrome methods. Results The menisci frozen for 1 month showed partially preserved collagen fiber structure and no significant hydropic tissue degeneration. The menisci frozen for 5 years presented an evident dissociation of collagen fibers and multiple foci of hydropic degeneration. Discussion Degeneration was much more significant in menisci stored for 5 years, indicating that a long freezing period results in substantial progression of tissue deterioration. This may suggest that the 5-year period, considered the maximum time for graft storage before transplant, is too long. Conclusion Grafts stored for 1 month showed a slight degenerative change in collagen fibers, whereas menisci frozen for 5 years presented significant tissue degeneration.


Resumo Objetivos Avaliar e comparar as características histológicas de meniscos frescos e meniscos congelados armazenados em banco de tecidos por 1 mês e por 5 anos. Métodos Foi feito um estudo histológico com enxertos meniscais. Avaliamos 10 meniscos, sendo 2 que ficaram armazenados sob congelamento por 5 anos, 4 armazenados congelados por 1 mês, e 4 frescos, recém captados. Foram feitos cortes histológicos corados com hematoxilina e eosina e Tricrômico de Masson, para avaliação das propriedades histológicas. Resultados Os meniscos congelados por 1 mês apresentaram preservação parcial da estrutura das fibras colágenas, sem degeneração hidrópica significativa do tecido. Nos meniscos congelados por 5 anos, observamos dissociação evidente das fibras colágenas, com presença de múltiplos focos de degeneração hidrópica. Discussão Encontramos degeneração bem mais significativa nos meniscos armazenados por 5 anos, o que indica que o longo período de congelamento leva à progressão significativa da degeneração do tecido. Isto pode sugerir que o período de 5 anos, considerado período máximo que o enxerto pode permanecer armazenado antes de ser transplantado, é um período muito longo. Conclusão Nos enxertos armazenados por 1 mês, existiu apenas discreta alteração degenerativa das fibras colágenas, enquanto que nos meniscos com 5 anos de congelamento foi observada degeneração significativa do tecido. Tibiais


Subject(s)
Tissue Banks , Wounds, Penetrating , Collagen , Eosine Yellowish-(YS) , Transplants , Meniscus , Freezing , Goals , Hematoxylin
3.
Rev. cuba. invest. bioméd ; 39(3): e640, jul.-set. 2020. tab, graf
Article in Spanish | LILACS, CUMED | ID: biblio-1138936

ABSTRACT

Introducción: La microscopía holográfica digital ha permitido a la microscopía óptica hacer uso de herramientas numéricas y computacionales; y esto, a su vez, ha favorecido múltiples avances en el estudio de las células y los tejidos en diferentes campos de la medicina y otras ciencias afines. Objetivo: Describir las características histológicas y morfométricas de los folículos tiroideos humanos con la microscopía holográfica digital. Métodos: Se realizó, desde el punto de vista histomorfométrico, un estudio descriptivo y transversal de folículos tiroideos humanos utilizando una instalación de microscopía holográfica digital. Se empleó la técnica de inclusión en parafina y tinción de hematoxilina-eosina para el procesamiento de las muestras. Se realizaron de 10 a 12 capturas de hologramas por muestra y el método de doble propagación para la reconstrucción de los hologramas. Se calculó el área, el perímetro, el diámetro mayor y menor de los folículos y cavidades foliculares y se realizaron reconstrucciones de imágenes holográficas en tres dimensiones. Se determinó como medida de tendencia central la media aritmética y como medida de dispersión la desviación típica o estándar. Resultados: Parámetros foliculares: área (5140,31 ± 1126,71 µm2); perímetro (2961,54 ± 71,2 µm); diámetro mayor:(921,17 ± 24,34 µm); diámetro menor: (746,67 ± 18,08 µm); altura del epitelio (7,92 ± 0,96). Cavidades foliculares: área (3686,18 ±1023,52 µm2); diámetro mayor: (698,86 ± 19,55 µm) y diámetro menor: (581,15 ± 13,82 µm). Conclusiones: Existen parámetros foliculares, determinados mediante la microscopía holográfica digital, no reportados por la literatura consultada, que resultan de interés en el estudio histológico de los folículos tiroideos humanos(AU)


Introduction: Digital holographic microscopy has made it possible to incorporate the use of numerical and computer tools into optical microscopy. This in turn has led to great progress in the study of cells and tissues in several fields of medicine and related sciences. Objective: Describe the histological and morphometric characteristics of human thyroid follicles using digital holographic microscopy. Methods: A descriptive cross-sectional histomorphometric study was conducted of human thyroid follicles using a digital holographic microscopy facility. Sample processing was based on inclusion technique by paraffin and hematoxylin-eosin staining. Ten to twelve holographic captures were made per sample, and the double propagation method was used for holographic reconstruction. Estimation was carried out of the area, perimeter, and greatest and smallest diameter of follicles and follicular cavities, and tri-dimensional reconstructions were made of holographic images. Arithmetic mean was determined as the measure of central tendency, and typical or standard deviation as the measure of dispersion. Results: Follicular parameters: area (5 140.31 ± 1 126.71 µm2); perimeter (2 961.54 ± 71.2 µm); greatest diameter (921.17 ± 24.34 µm); smallest diameter (746.67 ± 18.08 µm); epithelial height (7.92 ± 0.96). Follicular cavities: area (3 686.18 ± 1 023.52 µm2); greatest diameter (698.86 ± 19.55 µm); smallest diameter (581.15 ± 13.82 µm). Conclusions: A number of follicular parameters determined by digital holographic microscopy have not been reported by the literature consulted, and they are of interest to the histological study of human thyroid follicles(AU)


Subject(s)
Humans , Computers , Holography/methods , Hematoxylin/therapeutic use , Thyroid Gland/physiology , Eosine Yellowish-(YS)
4.
Arq. bras. med. vet. zootec. (Online) ; 72(3): 647-654, May-June, 2020. ilus, tab, graf
Article in English | LILACS, VETINDEX | ID: biblio-1128504

ABSTRACT

The elastic cartilage is composed by chondroblasts and chondrocytes, extracellular matrix and surrounded by perichondrium. It has a low regeneration capacity and is a challenge in surgical repair. One of obstacles in engineering a structurally sound and long-lasting tissue is selecting the most appropriate scaffold material. One of the techniques for obtaining biomaterials from animal tissues is the decellularization that decreases antigenicity. In this work, alkaline solution was used in bovine ear elastic cartilages to evaluate the decellularization and the architecture of the extracellular matrix. The cartilages were treated in alkaline solution (pH13) for 72 hours and lyophilized to be compared with untreated cartilages by histological analysis (hematoxylin-eosin, Masson's trichrome and Verhoeff slides). Areas of interest for cell counting and elastic fiber quantification were delineated, and the distribution of collagen and elastic fibers and the presence of non-fibrous proteins were observed. The results demonstrated that the alkaline solution caused 90% decellularization in the middle and 13% in the peripheral region, and maintenance of the histological characteristics of the collagen and elastic fibers and non-fibrous protein removal. It was concluded that the alkaline solution was efficient in the decellularization and removal of non-fibrous proteins from the elastic cartilages of the bovine ear.(AU)


A cartilagem elástica é composta por condroblastos e condrócitos, matriz extracelular e envolta por pericôndrio. Possui uma baixa capacidade de regeneração e é um desafio em reparos cirúrgicos. Um dos obstáculos na engenharia de tecido estruturalmente sólido e de longa duração é a seleção do material de arcabouço mais adequado. Uma das técnicas para obtenção de biomateriais oriundos de tecidos animais é a descelularização, que diminui a antigenicidade. Neste trabalho, foi utilizada solução alcalina em cartilagem elástica auricular bovina para avaliar a descelularização e a arquitetura da matriz extracelular. As cartilagens foram tratadas em solução alcalina (pH13) durante 72 horas e liofilizadas, e comparadas com cartilagens não tratadas por análise histológica (hematoxilina-eosina, tricrômio de Masson e Verhoeff). Foram determinadas as áreas de interesse para contagem celular e quantificação de fibras elásticas, observada a distribuição de colágeno e fibras elásticas e a presença de proteínas não fibrosas. Os resultados demonstraram que a solução alcalina causou 90% de descelularização na região central e 13% na região periférica, manutenção das características histológicas do colágeno e fibras elásticas e remoção das proteínas não fibrosas. Concluiu-se que a solução alcalina foi eficiente na descelularização e retirada de proteínas não fibrosas de cartilagens elásticas da orelha de bovinos.(AU)


Subject(s)
Biocompatible Materials , Chondrocytes , Tissue Engineering/veterinary , Elastic Cartilage , Extracellular Matrix , Cattle , Cartilage , Eosine Yellowish-(YS) , Alkalies
5.
Article in Chinese | WPRIM | ID: wpr-827542

ABSTRACT

OBJECTIVE@#To explore the treatment conditions of acid decalcified specimens and improve the poor quality of sections and unclear structure of hematoxylin-eosin (HE) staining caused by the change in pH in tooth and hard tissue after acid decalcification.@*METHODS@#A total of 20 cases of oral pathological specimens that contain hard tissues were decalcified and treated with routine treatment, concentrated ammonia water immersion treatment, and saturated lithium carbonate solution immersion treatment. The quality and HE staining effects of hard tissue sections treated with different methods were compared.@*RESULTS@#Compared with routine treatment, lithium carbonate saturated solution treatment showed complete sections. Hematoxylin is strongly stained, the nucleus is clear, and the cytoplasm is bright.@*CONCLUSIONS@#Soaking acid decalcified specimens in lithium carbonate saturated solution before embedding in dehydration can neutralize the acidic environment of the tissue. The quality of sections and HE staining effect are improved and are suitable for the pretreatment of acid decalcified tissue samples of oral pathology.


Subject(s)
Eosine Yellowish-(YS) , Hematoxylin , Staining and Labeling , Tooth
6.
Article in English | WPRIM | ID: wpr-742388

ABSTRACT

BACKGROUND: Tissue engineering is a multidisciplinary field which attracted much attention in recent years. One of the most important issue in tissue engineering is how to obtain high cell numbers and tissue regeneration while maintaining appropriate cellular characteristics in vitro for restoring damaged or dysfunctional body tissues and organs. These demands can be achieved by the use of three dimensional (3D) dynamic cultures of cells combined with cell-adhesive micro-carriers. METHODS: In this study, human mesenchymal stem cells (hMSCs) were cultured in a wave-bioreactor system for up to 100 days, after seeding on Cultisphere-S porous gelatin micro-carriers. Cell counting was performed at the time points of 7, 12, 17, 31 days and compared to those of hMSCs cultured under static condition. Higher growth and proliferation rates was achieved in wave-type dynamic culture, when cell culture continued to day 31. A scanning electron microscope (SEM) photographs, both live and dead and MTT assays were taken to confirm the survival and distribution of cells on porous gelatin micro-carrier surfaces. The results of histological stains such as hematoxylin and eosin, Masson’s trichrome, Alcian blue and Alizarin red S also showed improved proliferation and tissue regeneration of hMSCs on porous gelatin micro-carriers. CONCLUSION: The experimental results demonstrated the effect and importance of both micro-carriers and bioreactor in hMSC expansion on cell proliferation and migration as well as extracellular matrix formation on the superficial and pore surfaces of the porous gelatin micro-carriers, and then their inter-connections, leading to tissue regeneration.


Subject(s)
Alcian Blue , Bioreactors , Cell Count , Cell Culture Techniques , Cell Proliferation , Coloring Agents , Eosine Yellowish-(YS) , Extracellular Matrix , Gelatin , Hematoxylin , Humans , In Vitro Techniques , Mesenchymal Stem Cells , Regeneration , Tissue Engineering
7.
Article in English | WPRIM | ID: wpr-742383

ABSTRACT

BACKGROUND: Articular cartilage damage is still a troublesome problem. Hence, several researches have been performed for cartilage repair. The aim of this study was to evaluate the chondrogenicity of demineralized bone matrix (DBM) scaffolds under cyclic hydrostatic pressure (CHP) in vitro. METHODS: In this study, CHP was applied to human bone marrow mesenchymal stem cells (hBMSCs) seeded on DBM scaffolds at a pressure of 5 MPa with a frequency of 0.5 Hz and 4 h per day for 1 week. Changes in chondrogenic and osteogenic gene expressions were analyzed by quantifying mRNA signal level of Sox9, collagen type I, collagen type II, aggrecan (ACAN), Osteocalcin, and Runx2. Histological analysis was carried out by hematoxylin and eosin, and Alcian blue staining. Moreover, DMMB and immunofluorescence staining were used for glycosaminoglycan (GAG) and collagen type II detection, respectively. RESULTS: Real-time PCR demonstrated that applying CHP to hBMSCs in DBM scaffolds increased mRNA levels by 1.3-fold, 1.2-fold, and 1.7-fold (p < 0.005) for Sox9, Col2, and ACAN, respectively by day 21, whereas it decreased mRNA levels by 0.7-fold and 0.8-fold (p < 0.05) for Runx2 and osteocalcin, respectively. Additionally, in the presence of TGF-β1 growth factor (10 ng/ml), CHP further increased mRNA levels for the mentioned genes (Sox9, Col2, and ACAN) by 1.4-fold, 1.3-fold and 2.5-fold (p < 0.005), respectively. Furthermore, in histological assessment, it was observed that the extracellular matrix contained GAG and type II collagen in scaffolds under CHP and CHP with TGF-β1, respectively. CONCLUSION: The osteo-inductive DBM scaffolds showed chondrogenic characteristics under hydrostatic pressure. Our study can be a fundamental study for the use of DBM in articular cartilage defects in vivo and lead to production of novel scaffolds with two different characteristics to regenerate both bone and cartilage simultaneously.


Subject(s)
Aggrecans , Alcian Blue , Bone Marrow , Bone Matrix , Cartilage , Cartilage, Articular , Collagen Type I , Collagen Type II , Eosine Yellowish-(YS) , Extracellular Matrix , Fluorescent Antibody Technique , Gene Expression , Hematoxylin , Humans , Hydrostatic Pressure , In Vitro Techniques , Mesenchymal Stem Cells , Osteocalcin , Real-Time Polymerase Chain Reaction , RNA, Messenger
8.
Article in English | WPRIM | ID: wpr-741993

ABSTRACT

OBJECTIVES: To achieve pulp-dentin complex regeneration with tissue engineering, treatment efficacies and safeties should be evaluated using in vivo orthotopic transplantation in a sufficient number of animals. Mice have been a species of choice in which to study stem cell biology in mammals. However, most pulp-dentin complex regeneration studies have used large animals because the mouse tooth is too small. The purpose of this study was to demonstrate the utility of the mouse tooth as a transplantation model for pulp-dentin complex regeneration research. MATERIALS AND METHODS: Experiments were performed using 7-week-old male Institute of Cancer Research (ICR) mice; a total of 35 mice had their pulp exposed, and 5 mice each were sacrificed at 1, 2, 4, 7, 9, 12 and 14 days after pulp exposure. After decalcification in 5% ethylenediaminetetraacetic acid, the samples were embedded and cut with a microtome and then stained with hematoxylin and eosin. Slides were observed under a high-magnification light microscope. RESULTS: Until 1 week postoperatively, the tissue below the pulp chamber orifice appeared normal. The remaining coronal portion of the pulp tissue was inflammatory and necrotic. After 1 week postoperatively, inflammation and necrosis were apparent in the root canals inferior to the orifices. The specimens obtained after experimental day 14 showed necrosis of all tissue in the root canals. CONCLUSIONS: This study could provide opportunities for researchers performing in vivo orthotopic transplantation experiments with mice.


Subject(s)
Animals , Biology , Dental Pulp Cavity , Dental Pulp Necrosis , Edetic Acid , Eosine Yellowish-(YS) , Hematoxylin , Humans , Inflammation , Male , Mammals , Mice , Necrosis , Pulpitis , Regeneration , Safety , Stem Cells , Tissue Engineering , Tooth
9.
Anatomy & Cell Biology ; : 93-96, 2019.
Article in English | WPRIM | ID: wpr-738807

ABSTRACT

Thymus is an encapsulated organ having its bilateral origin from the third pharyngeal pouch. It appears to be a single organ but actually it is bilobed. It attains its maximum development at puberty and then it begins to involute. The parenchyma is replaced by adipocytes and lymphocyte production declines. Here we present a large thymus with a small area of persistent active tissue in it which was obtained during routine undergraduate dissection class. Tissues taken from different quadrants of the large thymic mass were processed, embedded in paraffin and sections were taken for hematoxylin and eosin staining which showed presence of thymic tissue in only one quadrant. Further sections from that quadrant was treated with cytokeratin to confirm its epithelial origin. Therefore knowledge of a large persistent thymus will be helpful to the radiologists and surgeons for making differential diagnosis and in avoiding unnecessary surgical intervention.


Subject(s)
Adipocytes , Adolescent , Aged , Cadaver , Diagnosis, Differential , Eosine Yellowish-(YS) , Hematoxylin , Humans , Immunohistochemistry , Keratins , Lymphocytes , Paraffin , Puberty , Surgeons , Thymus Gland
11.
Acta cir. bras ; 34(5): e201900504, 2019. tab, graf
Article in English | LILACS | ID: biblio-1010871

ABSTRACT

Abstract Purpose: To establish a new rat model, the pathogenesis of which is closer to the clinical occurrence of chronic obstructive jaundice with liver fibrosis. Methods: 90 SD rats were randomly divided into 3 groups. Group A common bile duct ligation, group B common bile duct injection compont and group C injection saline. The serum of three groups was extracted, and the liver function was detected by ELISA. HE staining, Masson staining and immunohistochemistry were used to detect liver pathology. Results: Group B showed a fluctuant development of jaundice, obstructive degree reached a peak at 2 weeks, and decreased from 3 weeks. HA, LA and PCIII were significantly higher than control group. 3 weeks after surgery, liver tissue fibrosis occurred in group B, and a wide range of fiber spacing was formed at 5 weeks. Immunohistochemistry showed that hepatic stellate cells were more active than the control group. Conclusion: Intra-biliary injection of Compont gel is different from the classic obstructive jaundice animal model caused by classic bile duct ligation, which can provide an ideal rat model of chronic obstructive jaundice with liver fibrosis.


Subject(s)
Animals , Female , Bile Ducts/drug effects , Disease Models, Animal , Gels/administration & dosage , Liver Cirrhosis/chemically induced , Aspartate Aminotransferases/blood , Reference Values , Azo Compounds , Time Factors , Bile Ducts/pathology , Bilirubin/analysis , Serum Albumin/analysis , Enzyme-Linked Immunosorbent Assay , Immunohistochemistry , Random Allocation , Reproducibility of Results , Rats, Sprague-Dawley , Eosine Yellowish-(YS) , Jaundice, Obstructive/chemically induced , Jaundice, Obstructive/pathology , Alkaline Phosphatase/blood , gamma-Glutamyltransferase/blood , Injections , Liver Cirrhosis/pathology , Methyl Green
12.
Article in English | WPRIM | ID: wpr-765938

ABSTRACT

BACKGROUND/AIMS: Functional dyspepsia (FD) is characterized as chronic recurrent upper gastrointestinal symptoms in the absence of any organic disorder. We hypothesized that duodenal low-grade inflammation activates superficial afferent nerve sprouting, thereby contributing to hypersensitivity in patients with FD. METHODS: A prospective case-control study was conducted in a tertiary referral center. FD was defined using the Rome III criteria. Standardized endoscopic biopsies were performed in the stomach and duodenum. Hematoxylin and eosin staining and immunohistochemical staining for major basic proteins were performed to detect granulated eosinophil-derived granules, and S-100 staining was performed to detect fine nerve fibers. RESULTS: A total of 51 patients with FD (82% female; mean age 35.8 ± 13.4 years) and 35 controls were enrolled. Activated eosinophil counts in the duodenum were significantly higher in patients with FD than in controls (41.4% vs 17.1%, P = 0.005). Microscopic duodenitis was more frequently detected in patients with FD than in controls. Fine nerve fibers were more abundant in patients with FD than in controls (45.1% vs 11.4%, P = 0.029). The abundance of fine nerve fibers highly correlated with the degree of activated eosinophils. CONCLUSION: Duodenal low-grade inflammation, such as mucosal eosinophilic accumulation with degranulation, promoted mucosal enteric nerve fiber density and sprouting in patients with FD.


Subject(s)
Biopsy , Case-Control Studies , Duodenitis , Duodenum , Dyspepsia , Eosine Yellowish-(YS) , Eosinophils , Female , Hematoxylin , Humans , Hypersensitivity , Inflammation , Mucous Membrane , Nerve Fibers , Peripheral Nervous System , Prospective Studies , Stomach , Tertiary Care Centers
13.
Article in English | WPRIM | ID: wpr-765625

ABSTRACT

STUDY DESIGN: Animal study. OBJECTIVES: To investigate the effects of microelectric treatment by transcutaneous electrical nerve stimulation (TENS) on functional recovery and histological changes in a rat model of spinal cord injury (SCI). SUMMARY OF LITERATURE REVIEW: The effects of TENS on spasticity and its underlying mechanisms remain unclear. MATERIALS AND METHODS: SCI was induced by a 1.5-mm impactor with 200,000–260,000 dyne after laminectomy. Rats were divided into the following groups: group I (normal control), group II (microelectric treatment of 0 A), group III (microelectric treatment of 100 µA for 1 hr/day), group IV (microelectric treatment of 400 µA for 1 hr/day), and group V (microelectric treatment of 400 µA for 24 hr/day). After inducing SCI, rats were assessed by a sensory test with von Frey filaments and the locomotor recovery test (BBB rating scale) at 1, 4, 7, 14, 21, and 28 days. To evaluate spinal cord damage, histopathological studies were performed with hematoxylin and eosin. Brain-derived neurotrophic factor (BDNF) and TrkB immunohistochemistry studies were performed at 28 days. RESULTS: In groups IV and V, the BBB score had significantly improved on days 21 and 28 after SCI, and the TENS-treated groups showed significant neuronal recovery. After SCI, groups IV and V showed a significant recovery of locomotor function and the motor sensory response of the withdrawal threshold to 3.5 g. In addition, necrotic tissue and cystic spaces in the spinal cord were significantly reduced and BDNF/TrkB-positive cells were highly expressed in groups III, IV, and V. CONCLUSIONS: Microelectric treatment can play a role in facilitating the recovery of locomotion following SCI.


Subject(s)
Animals , Brain-Derived Neurotrophic Factor , Eosine Yellowish-(YS) , Hematoxylin , Immunohistochemistry , Laminectomy , Locomotion , Models, Animal , Muscle Spasticity , Neurons , Rats , Spinal Cord Injuries , Spinal Cord , Transcutaneous Electric Nerve Stimulation
14.
Article in English | WPRIM | ID: wpr-764061

ABSTRACT

BACKGROUND AND OBJECTIVE: The characteristics of human hematopoietic stem cells are conditioned by the microenvironment of the bone marrow, where they interact with other cell populations, such as mesenchymal stem cells and endothelial cells; however, the study of this microenvironment is complex. The objective of this work was to develop a 3D culture system by magnetic levitation that imitates the microenvironment of human HSC. METHODS AND RESULTS: Human bone marrow-mesenchymal stem cells, umbilical cord blood-hematopoietic stem cells and a non-tumoral endothelial cell line (CC2811, Lonza®) were used to develop organotypic multicellular spheres by the magnetic levitation method. We obtained viable structures with an average sphericity index greater than 0.6, an average volume of 0.5 mm3 and a percentage of aggregation greater than 70%. Histological studies of the organotypic multicellular spheres used hematoxylin and eosin stains, and an evaluation of vimentin expression by means of immunohistochemistry demonstrated an organized internal structure without picnotic cells and a high expression of vimentin. The functional capacity of human hematopoietic stem cells after organotypic multicellular spheres culture was evaluated by multipotency tests, and it was demonstrated that 3D structures without exogenous Flt3L are autonomous in the maintenance of multipotency of human hematopoietic stem cells. CONCLUSIONS: We developed organotypic multicellular spheres from normal human cells that mimic the microenvironment of the human hematopoietic stem cells. These structures are the prototype for the development of complex organoids that allow the further study of the biology of normal human stem cells and their potential in regenerative medicine.


Subject(s)
Biology , Bone Marrow , Coloring Agents , Endothelial Cells , Eosine Yellowish-(YS) , Hematopoietic Stem Cells , Hematoxylin , Humans , Immunohistochemistry , Mesenchymal Stem Cells , Methods , Organoids , Regenerative Medicine , Stem Cells , Umbilical Cord , Vimentin
15.
Natural Product Sciences ; : 261-267, 2019.
Article in English | WPRIM | ID: wpr-760562

ABSTRACT

The rhizomes of Dioscorea japonica Thunb. are widely consumed as food and also used to treat diabetes and polyuria in Korea. This study was undertaken to study the anti-atopic dermatitis effects of a 95% ethanolic extract (DJE) of D. japonica in an oxazolone-stimulated murine model of atopic dermatitis (AD). The therapeutic effects of DJE on AD-like skin lesions were assessed on both ears. DJE (1%) or dexamethasone (0.5%; the positive control) were applied to skin lesions for three weeks. Serum levels of IgE and IL-4 were assessed by ELISA (enzyme-linked immunosorbent assay). Histopathological examinations were performed by hematoxylin and eosin (H&E) and toluidine blue staining and revealed DJE significantly reduced dermal thickness and inflammatory cell infiltration when applied to oxazolone-treated ear skin. DJE-treated AD mice also showed lower serum levels of IgE and IL-4 than oxazolone-stimulated controls. Our findings demonstrate DJE might be a useful safe, topical agent for the treatment of atopic diseases.


Subject(s)
Animals , Dermatitis , Dermatitis, Atopic , Dexamethasone , Dioscorea , Ear , Enzyme-Linked Immunosorbent Assay , Eosine Yellowish-(YS) , Ethanol , Hematoxylin , Immunoglobulin E , Interleukin-4 , Korea , Mice , Oxazolone , Polyuria , Rhizome , Skin , Therapeutic Uses , Tolonium Chloride
16.
Article in English | WPRIM | ID: wpr-760362

ABSTRACT

Autophagy is a fundamental cellular process that maintains homeostasis and cell integrity, under stress conditions. Although the involvement of autophagy in various conditions has been elucidated, the role of autophagy in renal structure is not completely clarified. Our aim was to investigate the cytoprotective effect of autophagy against acute kidney injury (AKI) through cisplatin deteriorative pathway, which leads to AKI via renal cell degradation. For in vivo experiments, male Sprague Dawley rats were divided in to 2 groups (n = 6/group) as control, Cis-5D. Following a single intraperitoneal injection of cisplatin, rats were sacrificed after 5 days. Blood urea nitrogen (BUN), creatinine (Cr) and histological alterations were examined. Further, expression of key regulators of autophagy, light-clain 3 (LC3), p62, and Beclin1, was evaluated by immunohistochemistry (IHC). The rats exhibited severe renal dysfunction, indicated by elevated BUN, Cr. Hematoxylin and eosin staining revealed histological damages in cisplatin-treated rats. Furthermore, IHC analysis revealed increased expression of LC3, Beclin1 and decreased expression of p62. Furthermore, expression of aforementioned autophagy markers was restricted to proximal tubule. Taken together, our study demonstrated that cisplatin can cause nephrotoxicity and lead to AKI. This phenomenon accelerated autophagy in renal proximal tubules and guards against AKI.


Subject(s)
Acute Kidney Injury , Animals , Autophagy , Blood Urea Nitrogen , Cisplatin , Creatinine , Eosine Yellowish-(YS) , Hematoxylin , Homeostasis , Humans , Immunohistochemistry , Injections, Intraperitoneal , Male , Rats , Rats, Sprague-Dawley
17.
Article in English | WPRIM | ID: wpr-785606

ABSTRACT

OBJECTIVE: Sequelae of behavioral impairments associated with human traumatic brain injury (TBI) include neurobehavioral problems. We compared exploratory, cognitive, and depressive-like behaviors in pediatric and adult male mice exposed to controlled cortical impact (CCI).METHODS: Pediatric (21 to 25 days old) and adult (8 to 12 weeks old) male C57Bl/6 mice underwent CCI at a 2-mm depth of deflection. Hematoxylin and eosin staining was performed 3 to 7 days after recovery from CCI, and injury volume was analyzed using ImageJ. Neurobehavioral characterization after CCI was performed using the Barnes maze test (BMT), passive avoidance test, open-field test, light/dark test, tail suspension test, and rotarod test. Acutely and subacutely (3 and 7 days after CCI, respectively), CCI mice showed graded injury compared to sham mice for all analyzed deflection depths.RESULTS: Time-dependent differences in injury volume were noted between 3 and 7 days following 2-mm TBI in adult mice. In the BMT, 2-mm TBI adults showed spatial memory deficits compared to sham adults (P < 0.05). However, no difference in spatial learning and memory was found between sham and 2-mm CCI groups among pediatric mice. The open-field test, light/dark test, and tail suspension test did not reveal differences in anxiety-like behaviors in both age groups.CONCLUSION: Our findings revealed a graded injury response in both age groups. The BMT was an efficient cognitive test for assessing spatial/non-spatial learning following CCI in adult mice; however, spatial learning impairments in pediatric mice could not be assessed.


Subject(s)
Adult , Animals , Brain Injuries , Eosine Yellowish-(YS) , Hematoxylin , Hindlimb Suspension , Humans , Learning , Male , Memory , Mice , Rotarod Performance Test , Spatial Learning , Spatial Memory
18.
Annals of Dermatology ; : 662-665, 2019.
Article in English | WPRIM | ID: wpr-762390

ABSTRACT

Hair follicle nevus (HFN) is a rare, benign, follicular hamartoma that most frequently presents as a congenital nodule on the face. We experienced a rare case of HFN on the neck of a 14-year-old boy and performed a pilot immunohistochemical study with cytokeratin 19 (CK19) to compare the staining pattern of hair follicles in HFN and its differential diagnoses, accessory tragus, cervical chondrocutaneous branchial remnants (CCBR) and trichofolliculoma. With hematoxylin and eosin stain, HFN showed numerous tiny hair follicles in the dermis with several sebaceous and eccrine glands, and perifollicular fibrous thickening. With CK19 stain, some hair follicles in HFN and CCBR showed positive expression, a few hair follicles in accessory tragus showed weak expression, and no hair follicles in trichofolliculoma showed expression. The present report supports the view that HFN, accessory tragus and CCBR are within the same spectrum of hamartomas.


Subject(s)
Adolescent , Dermis , Diagnosis, Differential , Eccrine Glands , Eosine Yellowish-(YS) , Hair Follicle , Hair , Hamartoma , Hematoxylin , Humans , Keratin-19 , Male , Neck , Nevus
19.
Annals of Coloproctology ; : 174-180, 2019.
Article in English | WPRIM | ID: wpr-762320

ABSTRACT

PURPOSE: Analysis of the sentinel lymph node (SLN) in colorectal cancer (CRC) patients was proposed for more accurate staging and tailored lymphadenectomy. The aim of this study was to assess the ability to predict lymph node (LN) involvement through analysis of the SLN with a one-step nucleic acid (OSNA) technique in combination with peritumoral injection of indocyanine green (ICG) and near-infrared (NIR) lymphangiography in CRC patients. METHODS: A total of 34 patients were enrolled. Overall, 51 LNs were analyzed with OSNA. LNs of 17 patients (50%) were examined simultaneously with hematoxylin and eosin (H&E) and OSNA. RESULTS: SLN analysis of 17 patients examined with H&E and OSNA revealed that OSNA had a higher sensitivity (1 vs. 0.55), higher negative predictive value (1 vs. 0.66) and higher accuracy (100% vs. 76.4%) in predicting LN involvement. Overall, OSNA showed a sensitivity of 0.69, specificity of 1, accuracy of 88.2%, and stage migration of 8.8%. Compared to those who were OSNA (−), OSNA (+) patients had a greater number of LN metastases (4.8 vs. 0.16, P = 0.04), higher G3 rate (44.4% vs. 4%, P = 0.01), more advanced stage of disease (stage III: 77.8% vs. 16%; P = 0.00) and were more rapidly subjected to adjuvant chemotherapy (39.1 days vs. 50.2 days, P = 0.01). CONCLUSION: SLN analysis with OSNA in combination with ICG-NIR lymphangiography is feasible and can detect LN involvement in CRC patients. Furthermore, it allows for more accurate staging reducing the delay between surgery and adjuvant chemotherapy.


Subject(s)
Chemotherapy, Adjuvant , Colorectal Neoplasms , Eosine Yellowish-(YS) , Fluorescence , Hematoxylin , Humans , Indocyanine Green , Lymph Node Excision , Lymph Nodes , Lymphography , Neoplasm Metastasis , Sensitivity and Specificity
20.
Anatomy & Cell Biology ; : 204-207, 2019.
Article in English | WPRIM | ID: wpr-762211

ABSTRACT

A 51-year-old male was routinely biopsied during a paraspinal muscle study. The biopsy sample was taken from the right erector spinae muscle at the fourth lumbar vertebra. The patient had no history of (diagnosed) major back trauma. The obtained sample was histologically analyzed (hematoxylin and eosin, safranin O), and complementary magnetic resonance imaging was performed. The biopsied sample contained chondroid tissue. Based on its location, the biopsy sample was appointed as chondroid metaplasia. Although chondroid metaplasia is not uncommon in humans, this is the first report of chondroid metaplasia within the paraspinal connective tissue. We propose a novel mechanism to explain the paraspinal chrondrogenic changes, related to spinal degeneration.


Subject(s)
Biopsy , Connective Tissue , Eosine Yellowish-(YS) , Humans , Magnetic Resonance Imaging , Male , Metaplasia , Middle Aged , Paraspinal Muscles , Spine
SELECTION OF CITATIONS
SEARCH DETAIL