RESUMEN
Ultrasound is widely used in clinical practice, mostly in diagnostic studies, but increasingly in therapeutic applications as well. This may be the case in acceleration of wound healing or treatment of cancer. Still, little is known about the direct effect of frequency or energy density of the ultrasound upon the cells themselves. We therefore investigated the impact of three different protocols using high, medium and low energy densities at three different frequencies on normal endothelial and epithelial as well as carcinoma cell lines (neuroblastoma and adenocarcinoma cell lines). Proliferation of endothelial and epithelial cell lines was significantly increased depending on the frequency and energy density applied. No influence on actin cytoskeleton formation was seen in these cells after treatment, while a significant decrease in the density of microvilli and the length of filopodia in the epithelial cell line could be noted. The proliferation rate of the carcinoma cell lines was reduced and cells destroyed. Apoptosis was induced in the adenocarcinoma cells after ultrasound exposure. Additionally, the expression of neurofilament was increased in neuroblastoma cells as evidence of beginning differentiation. So, different settings of frequency and energy density in an ultrasonic treatment protocol lead to different impacts on proliferation, morphology and differentiation and might be used to stimulate or inhibit the growth of individual cell types.
Asunto(s)
Células/efectos de la radiación , Ultrasonido , Animales , Apoptosis/efectos de la radiación , Diferenciación Celular/efectos de la radiación , Línea Celular , Línea Celular Tumoral , Proliferación Celular/efectos de la radiación , Perros , Relación Dosis-Respuesta en la Radiación , Electroforesis en Gel Bidimensional , Técnica del Anticuerpo Fluorescente , Humanos , Liposomas , Ratones , Microscopía Electrónica de Rastreo , Proteoma , Ultrasonido/instrumentaciónRESUMEN
A morphometric and immunohistochemical study was performed on trephine biopsies of the bone marrow in patients showing idiopathic thrombocytopenic purpura (ITP) and reactive thrombocytosis (RTH). Features of megakaryocytopoiesis were determined not only by using the periodic acid Schiff reaction (PAS), but also by immunostaining with the monoclonal antibody Y2/51, detecting a formalin-resistant epitope of glycoprotein IIIa (CD61). An increased number of megakaryocytes was observed in both ITP and in RTH as compared with a control group. Immunostaining revealed a predominance of small immature elements of this cell lineage, particularly in cases of ITP. Following previous experimental cell culture studies on megakaryocyte precursors and features derived from corresponding smear preparations, we characterized promegakaryoblasts immunomorphometrically. According to these calculations, ITP, as opposed to RTH, features a disproportionate expansion of the megakaryocyte precursor pool.
Asunto(s)
Médula Ósea/patología , Hematopoyesis , Megacariocitos/patología , Púrpura Trombocitopénica/patología , Anticuerpos Monoclonales , División Celular , Núcleo Celular/ultraestructura , Células Madre Hematopoyéticas/patología , Humanos , Reacción del Ácido Peryódico de Schiff , Glicoproteínas de Membrana Plaquetaria/inmunología , Trombocitosis/patologíaRESUMEN
To determine the number of megakaryocyte precursors (pro- and megakaryoblasts), an immunomorphometric study was performed on paraffin-embedded trephine biopsies of the bone marrow using a monoclonal antibody against platelet glycoprotein IIIa. Eighteen control specimens from patients with no evidence of any hematological disorder and a normal platelet count were selected and assessed together with the same number of specimens from patients with reactive thrombocytosis, polycythemia vera rubra (P. vera) or primary (essential) thrombocythemia (PTH). A strikingly proportionate increase in early megakaryocytes occurred in all patients enrolled in this study, compared with the controls. Moreover, there were no significant correlations between counts for precursors or total megakaryocytes per square millimeter of bone marrow with the corresponding values for platelets. This indicates that despite an orderly increase in immature forms in the bone marrow, the number of platelets circulating in the blood is influenced by other additional factors, such as the expanded platelet pool in the enlarged spleen. The non-disproportionate expansion of megakaryocyte precursors extends previous findings on progenitor cells of this lineage in vitro, particularly in PTH. Histological evaluation of the bone marrow of patients with P. vera and PTH indicated that megakaryopoiesis proceeded to the production of appropriate mature forms with no obvious excess of very small or blastic elements.