RESUMO
BACKGROUND: Patients with inflammatory bowel disease (IBD) have a higher risk of developing colitis-associated colorectal cancer (CAC) with poor prognosis. IBD etiology remains undefined but involves environmental factors, genetic predisposition, microbiota imbalance (dysbiosis) and mucosal immune defects. Mesenchymal stromal cell (MSC) injections have shown good efficacy in reducing intestinal inflammation in animal and human studies. However, their effect on tumor growth in CAC and their capacity to restore gut dysbiosis are not clear. METHODS: The outcome of systemic administrations of in vitro expanded human intestinal MSCs (iMSCs) on tumor growth in vivo was evaluated using the AOM/DSS model of CAC in C57BL/6J mice. Innate and adaptive immune responses in blood, mesenteric lymph nodes (MLNs) and colonic tissue were analyzed by flow cytometry. Intestinal microbiota composition was evaluated by 16S rRNA amplicon sequencing. RESULTS: iMSCs significantly inhibited colitis and intestinal tumor development, reducing IL-6 and COX-2 expression, and IL-6/STAT3 and PI3K/Akt signaling. iMSCs decreased colonic immune cell infiltration, and partly restored intestinal monocyte homing and differentiation. iMSC administration increased the numbers of Tregs and IFN-γ+CD8+ T cells in the MLNs while decreasing the IL-4+Th2 response. It also ameliorated intestinal dysbiosis in CAC mice, increasing diversity and Bacillota/Bacteroidota ratio, as well as Akkermansia abundance, while reducing Alistipes and Turicibacter, genera associated with inflammation. CONCLUSION: Administration of iMSCs protects against CAC, ameliorating colitis and partially reverting intestinal dysbiosis, supporting the use of MSCs for the treatment of IBD.
Assuntos
Neoplasias Associadas a Colite , Colite , Doenças Inflamatórias Intestinais , Células-Tronco Mesenquimais , Humanos , Camundongos , Animais , Neoplasias Associadas a Colite/complicações , Neoplasias Associadas a Colite/patologia , Interleucina-6 , Camundongos Endogâmicos C57BL , Disbiose/complicações , Linfócitos T CD8-Positivos , RNA Ribossômico 16S , Fosfatidilinositol 3-Quinases , Colite/patologia , Inflamação , Colo/patologia , Doenças Inflamatórias Intestinais/patologia , Imunidade , Sulfato de Dextrana , Modelos Animais de DoençasRESUMO
BACKGROUND: and Purpose: Colorectal cancer (CRC) is one of the cancers with the highest incidence in which APC gene mutations occur in almost 80% of patients. This mutation leads to ß-catenin aberrant accumulation and an uncontrolled proliferation. Apoptosis evasion, changes in the immune response and microbiota composition are also events that arise in CRC. Tetracyclines are drugs with proven antibiotic and immunomodulatory properties that have shown cytotoxic activity against different tumor cell lines. EXPERIMENTAL APPROACH: The effect of tigecycline was evaluated in vitro in HCT116 cells and in vivo in a colitis-associated colorectal cancer (CAC) murine model. 5-fluorouracil was assayed as positive control in both studies. KEY RESULTS: Tigecycline showed an antiproliferative activity targeting the Wnt/ß-catenin pathway and downregulating STAT3. Moreover, tigecycline induced apoptosis through extrinsic, intrinsic and endoplasmic reticulum pathways converging on an increase of CASP7 levels. Furthermore, tigecycline modulated the immune response in CAC, reducing the cancer-associated inflammation through downregulation of cytokines expression. Additionally, tigecycline favored the cytotoxic activity of cytotoxic T lymphocytes (CTLs), one of the main immune defenses against tumor cells. Lastly, the antibiotic reestablished the gut dysbiosis in CAC mice increasing the abundance of bacterial genera and species, such as Akkermansia and Parabacteroides distasonis, that act as protectors against tumor development. These findings resulted in a reduction of the number of tumors and an amelioration of the tumorigenesis process in CAC. CONCLUSION AND IMPLICATIONS: Tigecycline exerts a beneficial effect against CRC supporting the use of this antibiotic for the treatment of this disease.
Assuntos
Antineoplásicos , Neoplasias Colorretais , Animais , Camundongos , Tigeciclina/efeitos adversos , beta Catenina/metabolismo , Neoplasias Colorretais/genética , Carcinogênese , Transformação Celular Neoplásica/metabolismo , Via de Sinalização Wnt , Antineoplásicos/efeitos adversos , Imunidade , Antibacterianos/efeitos adversos , Proliferação de CélulasRESUMO
The purpose of this study was to analyze the association between components of the diet, metabolic risks, and the serum concentrations of adiponectin and interleukin-6 (IL-6). With prior informed consent, an analytical cross-sectional study was carried out with 72 students in their first year of university. The subjects had a mean age of 19.2 ± 1.0 years and body mass index of 23.38 ± 4.2, and they were mainly women (80.6%). Sociodemographic, anthropometric, and dietary data and metabolic risk factors were evaluated, and biochemical parameters and adipocytokines were also considered. The data were analyzed using means, ranges, and correlations, as well as principal components. In general, the protein, fat, and sodium intake were higher than the international dietary recommendations, and deficiencies in vitamins B5 and E, potassium, phosphorus, selenium, and zinc were observed. The most frequently observed metabolic risks were insulin resistance and hypoalphalipoproteinemia. IL-6 was positively correlated with lipid and protein intake. Adiponectin showed a positive correlation with high-density lipoprotein and a negative correlation with insulin, weight, and waist, while the adiponectin pattern was similar to that of vitamins E and A, which decreased with increasing intake of calories, macronutrients, and sodium. In general, a hypercaloric diet that was high in protein, fat, and sodium and deficient in vitamins, mainly fat-soluble, was associated with a lower concentration of adiponectin and a higher concentration of IL-6, which favor the presence of metabolic risks, including insulin resistance. Intervention studies are required to evaluate the dietary intake of metabolic markers in young people without comorbidities, which will lay the foundation for implementing prevention strategies.
Assuntos
Ingestão de Energia , Universidades , Adiponectina , Adolescente , Adulto , Índice de Massa Corporal , Estudos Transversais , Feminino , Humanos , Estudantes , Adulto JovemRESUMO
Recent advances in the application of bone marrow mesenchymal stem cells (BMMSC) for the treatment of tendon and ligament injuries in the horse suggest improved outcome measures in both experimental and clinical studies. Although the BMMSC are implanted into the tendon lesion in large numbers (usually 10 - 20 million cells), only a relatively small number survive (<10%) although these can persist for up to 5 months after implantation. This appears to be a common observation in other species where BMMSC have been implanted into other tissues and it is important to understand when this loss occurs, how many survive the initial implantation process and whether the cells are cleared into other organs. Tracking the fate of the cells can be achieved by radiolabeling the BMMSC prior to implantation which allows non-invasive in vivo imaging of cell location and quantification of cell numbers. This protocol describes a cell labeling procedure that uses Technetium-99m (Tc-99m), and tracking of these cells following implantation into injured flexor tendons in horses. Tc-99m is a short-lived (t1/2 of 6.01 hr) isotope that emits gamma rays and can be internalized by cells in the presence of the lipophilic compound hexamethylpropyleneamine oxime (HMPAO). These properties make it ideal for use in nuclear medicine clinics for the diagnosis of many different diseases. The fate of the labeled cells can be followed in the short term (up to 36 hr) by gamma scintigraphy to quantify both the number of cells retained in the lesion and distribution of the cells into lungs, thyroid and other organs. This technique is adapted from the labeling of blood leukocytes and could be utilized to image implanted BMMSC in other organs.
Assuntos
Células da Medula Óssea/diagnóstico por imagem , Doenças dos Cavalos/diagnóstico por imagem , Transplante de Células-Tronco Mesenquimais/veterinária , Células-Tronco Mesenquimais/diagnóstico por imagem , Tecnécio/química , Tendinopatia/veterinária , Tendões/diagnóstico por imagem , Animais , Células da Medula Óssea/patologia , Doenças dos Cavalos/patologia , Doenças dos Cavalos/terapia , Cavalos , Transplante de Células-Tronco Mesenquimais/métodos , Células-Tronco Mesenquimais/patologia , Cintilografia/métodos , Cintilografia/veterinária , Tecnécio/análise , Tendinopatia/diagnóstico por imagem , Tendinopatia/terapia , Tendões/patologiaRESUMO
Osteogenesis imperfecta (OI) types V and VI are caused, respectively, by a unique dominant mutation in IFITM5, encoding BRIL, a transmembrane ifitm-like protein most strongly expressed in the skeletal system, and recessive null mutations in SERPINF1, encoding pigment epithelium-derived factor (PEDF). We identified a 25-year-old woman with severe OI whose dermal fibroblasts and cultured osteoblasts displayed minimal secretion of PEDF, but whose serum PEDF level was in the normal range. SERPINF1 sequences were normal despite bone histomorphometry consistent with type VI OI and elevated childhood serum alkaline phosphatase. We performed exome sequencing on the proband, both parents, and an unaffected sibling. IFITM5 emerged as the candidate gene from bioinformatics analysis, and was corroborated by membership in a murine bone co-expression network module containing all currently known OI genes. The de novo IFITM5 mutation was confirmed in one allele of the proband, resulting in a p.S40L substitution in the intracellular domain of BRIL but was absent in unaffected family members. IFITM5 expression was normal in proband fibroblasts and osteoblasts, and BRIL protein level was similar to control in differentiated proband osteoblasts on Western blot and in permeabilized mutant osteoblasts by microscopy. In contrast, SERPINF1 expression was decreased in proband osteoblasts; PEDF was barely detectable in conditioned media of proband cells. Expression and secretion of type I collagen was similarly decreased in proband osteoblasts; the expression pattern of several osteoblast markers largely overlapped reported values from cells with a primary PEDF defect. In contrast, osteoblasts from a typical case of type V OI, with an activating mutation at the 5'-terminus of BRIL, have increased SERPINF1 expression and PEDF secretion during osteoblast differentiation. Together, these data suggest that BRIL and PEDF have a relationship that connects the genes for types V and VI OI and their roles in bone mineralization.
Assuntos
Proteínas do Olho/biossíntese , Proteínas de Membrana/genética , Mutação/genética , Fatores de Crescimento Neural/biossíntese , Osteoblastos/metabolismo , Osteogênese Imperfeita/genética , Serpinas/biossíntese , Adulto , Fosfatase Alcalina/metabolismo , Sequência de Aminoácidos , Sequência de Bases , Diferenciação Celular/genética , Criança , Pré-Escolar , Colágeno Tipo I/metabolismo , DNA Complementar/genética , Exoma/genética , Proteínas do Olho/metabolismo , Feminino , Redes Reguladoras de Genes , Humanos , Proteínas de Membrana/química , Dados de Sequência Molecular , Fatores de Crescimento Neural/metabolismo , Osteocalcina/metabolismo , Osteogênese Imperfeita/diagnóstico por imagem , Estabilidade Proteica , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Radiografia , Reprodutibilidade dos Testes , Análise de Sequência de DNA , Serpinas/metabolismo , Adulto JovemRESUMO
INTRODUCTION: Histologic studies of teeth from animal models of revascularization/revitalization are available; however, specimens from human studies are lacking. The nature of tissues formed in the canal of human revascularized/revitalized teeth was not well established. METHODS: An immature mandibular premolar with infected necrotic pulp and a chronic apical abscess was treated with revascularization/revitalization procedures. At both the 18-month and 2-year follow-up visits, radiographic examination showed complete resolution of the periapical lesion, narrowing of the root apex without root lengthening, and minimal thickening of the canal walls. The revascularized/revitalized tooth was removed because of orthodontic treatment and processed for histologic examination. RESULTS: The large canal space of revascularized/revitalized tooth was not empty and filled with fibrous connective tissue. The apical closure was caused by cementum deposition without dentin. Some cementum-like tissue was formed on the canal dentin walls. Inflammatory cells were observed in the coronal and middle third of revascularized/revitalized tissue. CONCLUSIONS: In the present case, the tissue formed in the canal of a human revascularized/revitalized tooth was soft connective tissue similar to that in the periodontal ligament and cementum-like or bone-like hard tissue, which is comparable with the histology observed in the canals of teeth from animal models of revascularization/revitalization.
Assuntos
Apexificação/métodos , Dente Pré-Molar/patologia , Abscesso Periapical/terapia , Compostos de Alumínio/uso terapêutico , Antibacterianos/administração & dosagem , Coagulação Sanguínea/fisiologia , Calcificação Fisiológica/fisiologia , Compostos de Cálcio/uso terapêutico , Cementogênese/fisiologia , Criança , Ciprofloxacina/administração & dosagem , Tecido Conjuntivo/patologia , Fístula Dentária/terapia , Cavidade Pulpar/patologia , Necrose da Polpa Dentária/terapia , Dentina/patologia , Combinação de Medicamentos , Feminino , Seguimentos , Humanos , Metilmetacrilatos/uso terapêutico , Metronidazol/administração & dosagem , Minociclina/administração & dosagem , Óxidos/uso terapêutico , Materiais Restauradores do Canal Radicular/uso terapêutico , Preparo de Canal Radicular/métodos , Silicatos/uso terapêutico , Ápice Dentário/patologia , Cimento de Óxido de Zinco e Eugenol/uso terapêuticoRESUMO
This study aimed to investigate immediate cell survival and distribution following different administration routes of mesenchymal stem cells (MSCs) into naturally occurring tendon injuries. Ten million MSCs, labeled with technetium-99m hexamethylpropyleneamine oxime, were implanted into 13 horses with naturally occurring tendon or ligament injuries intra-lesionally, intravenously and by regional perfusion, and traced for up to 48 h using planar gamma scintigraphy. Labeling efficiencies varied between 1.8% and 18.5% (mean 9.3%). Cells were retained in the damaged area after intra-lesional administration but only 24% of cells were still present within the tendon after 24 h. After intravenous injection, cells largely distributed to the lung fields, with no detectable cells in the tendon lesions. Significant labeling of the tendon lesions was observed in 11/12 horses following regional perfusion but at a lower level to intra-lesional injection. The highest cell numbers were retained after intra-lesional injection, although with considerable cell loss, while regional perfusion may be a viable alternative for MSC delivery. Cells did not "home" to damaged tendon in large numbers after intravenous administration. Cells were detected in the lungs most frequently after intravascular administration, although with no adverse effects. Low cell retention has important implications for designing effective clinical therapies for human clinical use.
Assuntos
Doenças dos Cavalos/terapia , Transplante de Células-Tronco Mesenquimais/métodos , Células-Tronco Mesenquimais/metabolismo , Tecnécio/metabolismo , Tendinopatia/veterinária , Traumatismos dos Tendões/veterinária , Tendões/metabolismo , Animais , Células Cultivadas , Feminino , Doenças dos Cavalos/patologia , Cavalos , Injeções Intralesionais , Injeções Intravenosas , Masculino , Células-Tronco Mesenquimais/citologia , Cintilografia , Tendinopatia/patologia , Tendinopatia/terapia , Traumatismos dos Tendões/patologia , Traumatismos dos Tendões/terapia , Tendões/citologia , Fatores de Tempo , Resultado do TratamentoRESUMO
Se estudiaron 100 recién nacidos de muy bajo peso (RNMBP) sobrevivientes, nacidos entre 1986-89 en el Hospital Edgardo Rebagliati Martins, IPSS, Lima-Perú, alimentados con leche materna y fórmula para prematuros suplementada con 1,000 UI de vitamina D, en los cuales se evaluaron ocho indicadores nutricionales. Estudiando la relación variable peso al nacimiento y tres variables dependientes (Indicadores Nutricionales): día de máxima pérdida de peso, número de días para recuperar el peso al nacimiento, y número de días para alcanzar 2,000 g; y con el uso del análisis de Regresión Múltiple se confeccionó una Curva de Crecimiento intrahospitalario. Se encontró que la ganancia de peso (g/día) era similar entre los niños de nuestro estudio y el reportado en estudios efectuados en países desarrollados a paratir de los 1,350 g. Los indicadores nutricionales son excelentes parámetros para estudiar el patrón evolutivo nutricional por ser fáciles de reproducir, confiables y accesibles al uso diario en las nurserías.