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      Bovine Bone Matrix Action Associated With Morphogenetic Protein in Bone Defects in Rats Submitted to Alcoholism Translated title: Acción de la Matriz Ósea Bovina Asociada a la Proteína Morfogenética en Defectos Óseos en Ratones Sometidos a Alcoholismo

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          Abstract

          Extended excessive alcohol use causes changes in bone tissue, thus affecting osteogenesis. The objective of this study was to evaluate if demineralized bone matrix (Gen-ox®) associated with bone morphogenetic protein (Gen-pro®) changes bone neoformation in rats submitted to experimental alcoholism. Forty male rats (Rattus norvegicus) were separated into 2 groups of 20 animals each: Group E1, which received ethyl alcohol at 25% and had the surgical cavity filled in only with blood clot; and Group E2, which received ethyl alcohol at 25% and had the surgical cavity filled in with demineralized bovine cortical bone associated with bone morphogenetic protein. The animals were submitted to a three-week period of gradual adaptation to alcohol, and then continued receiving alcohol at 25% for 90 days, when the surgical cavity was made. After the surgery, the animals continued consuming alcohol until reaching the sacrifice periods of 10, 20, 40, and 60 days, when the tibias were removed for histological processing. Results showed that surgical cavity repair and bone marrow reorganization occurred faster in Group E1 than in Group E2. At the end of the experiment, it was observed that animals in Group E2 had thick bony trabeculae surrounding the implanted material particles and a small area of connective tissue in the surface region. In conclusion, the implanted material did not accelerate bone neoformation, rather it served as a structure for osteogenesis.

          Translated abstract

          El abuso prolongado del alcohol produce alteraciones en el tejido óseo, interfiriendo en el proceso de la osteogénesis. El estudio tiene como objetivo evaluar si la matriz ósea bovina desmineralizada (Gen-ox®) asociada a la proteína morfogenética ósea (Gen-pro®) altera la neoformación ósea en ratones sometidos a alcoholismo experimental. Fueron utilizados 40 ratones machos (Rattus norvegicus), separados en dos grupos de 20 animales cada uno: Grupo E1, que recibió alcohol etílico a 25% con cavidad quirúrgica rellenada solamente por coágulo sanguíneo, y Grupo E2, que recibió sólo alcohol etílico a 25% con cavidad quirúrgica rellenada con hueso bovino desmineralizado cortical asociado a proteína morfogenética ósea. Después de 3 semanas de adaptación gradual al alcohol, los animales continuaron recibiéndolo en concentración de 25% por 90 días, cuando fue realizada la cavidad quirúrgica. Luego de la cirugía, los animales continuaron la ingestión alcohólica hasta los períodos de sacrificio de 10, 20, 40 y 60 días, cuando las tibias fueron removidas para su procesamiento histológico. Los resultados mostraron que en el Grupo E1 hubo reparación de la cavidad quirúrgica y reorganización de la médula ósea en un menor lapso temporal que en el Grupo E2. En el período final del experimento, se observó en los animales del Grupo E2 la presencia de trabéculas óseas espesas alrededor de las partículas de material implantado y pequeña área de tejido conjuntivo en la región superficial. Se puede concluir en que el material implantado no aceleró el proceso de neoformación ósea, sirviendo como estructura de base para generar osteogénesis.

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          Alcohol exposure and mechanisms of tissue injury and repair.

          Tissue injury owing to acute and chronic alcohol consumption has extensive medical consequences, with the level and duration of alcohol exposure affecting both the magnitude of injury and the time frame to recovery. While the understanding of many of the molecular processes disrupted by alcohol has advanced, mechanisms of alcohol-induced tissue injury remain a subject of intensive research. Alcohol has multiple targets, as it affects diverse cellular and molecular processes. Some mechanisms of tissue damage as a result of alcohol may be common to many tissue types, while others are likely to be tissue specific. Here, we present a discussion of the alcohol-induced molecular and cellular disruptions associated with injury or recovery from injury in bone, muscle, skin, and gastric mucosa. In every case, the goal of characterizing the sites of alcohol action is to devise potential measures for protection, prevention, or therapeutic intervention.
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            Inhibitory effect of alcohol on osteogenic differentiation in human bone marrow-derived mesenchymal stem cells.

            Alcohol-induced osteoporosis is characterized by a considerable suppression of osteogenesis. The objective of this investigation was to determine the effect of alcohol on gene expression, protein synthesis, and mineralization in human bone marrow-derived mesenchymal stem cells induced toward osteogenic differentiation in vitro. Human bone marrow-derived mesenchymal stem cells induced toward osteogenesis were cultured in the presence or absence of 50 mM alcohol. Stem cells were characterized by using SH2 antibody to the cell-surface antigen CD105/endoglin, and their proliferation in the presence of alcohol was quantified. The expression of genes for early, middle, and late markers of the osteogenic lineage was quantified by Northern analysis, and bone matrix protein synthesis was assayed. The effect of alcohol on cell-mediated matrix mineralization in terminally differentiated cultures was determined by von Kossa staining. Fluorescence-activated cell sorting analysis of human mesenchymal stem cells separated with a Percoll gradient proved 99% homogeneity by using SH2 antibody to the surface antigen CD105. Dose-dependent inhibition of proliferation of these stem cells occurred at concentrations greater than 50 mM alcohol. Gene expression of osteoblast-specific factor 2/core binding factor a1 (Osf2/Cbfa1), type I collagen, alkaline phosphatase, and osteocalcin (early, middle, and late markers for osteogenesis, respectively) was analyzed with and without osteogenic induction and treatment with 50 mM alcohol. After induction, Osf2/Cbfa1 levels were unresponsive to alcohol. To determine the effect of alcohol on human mesenchymal stem cell progression along the osteogenic pathway, messenger RNA (mRNA) levels for type I collagen, alkaline phosphatase, and osteocalcin were examined after osteogenic induction. After osteogenic induction, alcohol down-regulated the gene expression of type I collagen and significantly reduced its synthesis. Alcohol did not alter mRNA expression of alkaline phosphatase, a midstage marker for osteogenesis, but significantly decreased its activity compared with osteogenic induction alone. After induction, osteocalcin remained unchanged by alcohol at both the mRNA and protein levels. Histochemistry revealed decreased alkaline phosphatase staining and fewer alkaline phosphatase-positive cells in alcohol-treated human mesenchymal stem cell cultures. von Kossa staining revealed a reduction in the number of mineralizing nodules in stem cell cultures after alcohol treatment. Collectively, the data suggest that alcohol alters osteogenic differentiation in human bone marrow-derived mesenchymal stem cell cultures during lineage progression and provide further insight into alcohol-induced reduced bone formation.
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              What is the interest of normal and pathological morphological research to be quantitative: The exemple of the stereology

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                Author and article information

                Journal
                ijmorphol
                International Journal of Morphology
                Int. J. Morphol.
                Sociedad Chilena de Anatomía (Temuco, , Chile )
                0717-9502
                March 2012
                : 30
                : 1
                : 266-271
                Affiliations
                [02] Marília SP orgnameUniversity of Marília Brazil
                [03] Bauru SP orgnameSacred Heart University Brazil
                [01] Bauru SP orgnameUniversity of São Paulo orgdiv1Bauru School of Dentistry orgdiv2Departament of Biological Science Brazil
                Article
                S0717-95022012000100048 S0717-9502(12)03000148
                e9669d35-6417-49ab-9e60-9137265dd46d

                This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

                History
                : 20 July 2011
                : 21 November 2011
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 22, Pages: 6
                Product

                SciELO Chile


                Injerto xenogénico,Bone repair,Alcoholism,Xenograft,Bone morphogenetic proteins,Reparación ósea,Alcoholismo,Proteína morfogenética ósea

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