984 resultados para Autologous endometrial coculture


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Waddlia chondrophila, an obligate intracellular bacterium belonging to the Chlamydiales order, is considered as an emerging pathogen. Some clinical studies highlighted a possible role of W. chondrophila in bronchiolitis, pneumonia and miscarriage. This pathogenic potential is further supported by the ability of W. chondrophila to infect and replicate within human pneumocytes, macrophages and endometrial cells. Considering that W. chondrophila might be a causative agent of respiratory tract infection, we developed a mouse model of respiratory tract infection to get insight into the pathogenesis of W. chondrophila. Following intranasal inoculation of 2 x 108 W. chondrophila, mice lost up to 40% of their body weight, and succumbed rapidly from infection with a death rate reaching 50% at day 4 post-inoculation. Bacterial loads, estimated by qPCR, increased from day 0 to day 3 post-infection and decreased thereafter in surviving mice. Bacterial growth was confirmed by detecting dividing bacteria using electron microscopy, and living bacteria were isolated from lungs 14 days post-infection. Immunohistochemistry and histopathology of infected lungs revealed the presence of bacteria associated with pneumonia characterized by an important multifocal inflammation. The high inflammatory score in the lungs was associated with the presence of pro-inflammatory cytokines in both serum and lungs at day 3 post-infection. This animal model supports the role of W. chondrophila as an agent of respiratory tract infection, and will help understanding the pathogenesis of this strict intracellular bacterium.

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Immunotherapy is emerging as a promising anti-cancer curative modality. However, in contrast to recent advances obtained employing checkpoint blockade agents and T cell therapies, clinical efficacy of therapeutic cancer vaccines is still limited. Most vaccination attempts in the clinic represent "off-the shelf" approaches since they target common "self" tumor antigens, shared among different patients. In contrast, personalized approaches of vaccination are tailor-made for each patient and in spite being laborious, hold great potential. Recent technical advancement enabled the first steps in the clinic of personalized vaccines that target patient-specific mutated neo-antigens. Such vaccines could induce enhanced tumor-specific immune response since neo-antigens are mutation-derived antigens that can be recognized by high affinity T cells, not limited by central tolerance. Alternatively, the use of personalized vaccines based on whole autologous tumor cells, overcome the need for the identification of specific tumor antigens. Whole autologous tumor cells could be administered alone, pulsed on dendritic cells as lysate, DNA, RNA or delivered to dendritic cells in-vivo through encapsulation in nanoparticle vehicles. Such vaccines may provide a source for the full repertoire of the patient-specific tumor antigens, including its private neo-antigens. Furthermore, combining next-generation personalized vaccination with other immunotherapy modalities might be the key for achieving significant therapeutic outcome.

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Co-culture techniques associating both dermal fibroblasts and epidermal keratinocytes have shown to have better clinical outcome than keratinocyte culture alone for the treatment of severe burns. Since fat grafting has been shown to improve scar remodelling, new techniques such as cell-therapy-assisted surgical reconstruction with isolated and expanded autologous adipose-derived stem cells (ASCs) would be of benefit to increase graft acceptation. Therefore, integrating ASCs into standardized procedures for cultured skin grafting could be of benefit for the patient if cell quality and quantity could be maintained. The purpose of this study was to evaluate ASC processing from adult tissue with simple isolation (without enzymatic steps), expansion (low density of 325-3,000 cells/cm2) and storage conditions to assure methods to enhance the cellular resistance when transferred back to the patient. Co-culture with cell-banked skin progenitor cells (FE002-SK2) showed an increase of 40-50% ASCs yield at high passages alongside with a better preservation of morphology, proper adipogenic and osteogenic differentiation and efficient biocompatibility with 3D collagen scaffolds. ASCs can be considered as a valuable additional cell source to be delivered in biological bandages to the patient in a need of tissue reconstruction such as burn patients.

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Hormone-dependent diseases, e.g. cancers, rank high in mortality in the modern world, and thus, there is an urgent need for new drugs to treat these diseases. Although the diseases are clearly hormone-dependent, changes in circulating hormone concentrations do not explain all the pathological processes observed in the diseased tissues. A more inclusive explanation is provided by intracrinology – a regulation of hormone concentrations at the target tissue level. This is mediated by the expression of a pattern of steroid-activating and -inactivating enzymes in steroid target tissues, thus enabling a concentration gradient between the blood circulation and the tissue. Hydroxysteroid (17beta) dehydrogenases (HSD17Bs) form a family of enzymes that catalyze the conversion between low active 17-ketosteroids and highly active 17beta-hydroxysteroids. HSD17B1 converts low active estrogen (E1) to highly active estradiol (E2) with high catalytic efficiency, and altered HSD17B1 expression has been associated with several hormone-dependent diseases, including breast cancer, endometriosis, endometrial hyperplasia and cancer, and ovarian epithelial cancer. Because of its putative role in E2 biosynthesis in ovaries and peripheral target tissues, HSD17B1 is considered to be a promising drug target for estrogen-dependent diseases. A few studies have indicated that the enzyme also has androgenic activity, but they have been ignored. In the present study, transgenic mice overexpressing human HSD17B1 (HSD17B1TG mice) were used to study the effects of the enzyme in vivo. Firstly, the substrate specificity of human HSD17B1 was determined in vivo. The results indicated that human HSD17B1 has significant androgenic activity in female mice in vivo, which resulted in increased fetal testosterone concentration and female disorder of sexual development appearing as masculinized phenotype (increased anogenital distance, lack of nipples, lack of vaginal opening, combination of vagina with urethra, enlarged Wolffian duct remnants in the mesovarium and enlarged female prostate). Fetal androgen exposure has been linked to polycystic ovary syndrome (PCOS) and metabolic syndrome during adulthood in experimental animals and humans, but the genes involved in PCOS are largely unknown. A putative mechanism to accumulate androgens during fetal life by HSD17B1 overexpression was shown in the present study. Furthermore, as a result of prenatal androgen exposure locally in the ovaries, HSD17B1TG females developed ovarian benign serous cystadenomas in adulthood. These benign lesions are precursors of low-grade ovarian serous tumors. Ovarian cancer ranks fifth in mortality of all female cancers in Finland, and most of the ovarian cancers arise from the surface epithelium. The formation of the lesions was prevented by prenatal antiandrogen treatment and by transplanting wild type (WT) ovaries prepubertally into HSD17B1TG females. The results obtained in our non-clinical TG mouse model, together with a literature analysis, suggest that HSD17B1 has a role in ovarian epithelial carcinogenesis, and especially in the development of serous tumors. The role of androgens in ovarian carcinogenesis is considered controversial, but the present study provides further evidence for the androgen hypothesis. Moreover, it directly links HSD17B1-induced prenatal androgen exposure to ovarian epithelial carcinogenesis in mice. As expected, significant estrogenic activity was also detected for human HSD17B1. HSD17B1TG mice had enhanced peripheral conversion of E1 to E2 in a variety of target tissues, including the uterus. Furthermore, this activity was significantly decreased by treatments with specific HSD17B1 inhibitors. As a result, several estrogen-dependent disorders were found in HSD17B1TG females. Here we report that HSD17B1TG mice invariably developed endometrial hyperplasia and failed to ovulate in adulthood. As in humans, endometrial hyperplasia in HSD17B1TG females was reversible upon ovulation induction, triggering a rise in circulating progesterone levels, and in response to exogenous progestins. Remarkably, treatment with a HSD17B1 inhibitor failed to restore ovulation, yet completely reversed the hyperplastic morphology of epithelial cells in the glandular compartment. We also demonstrate that HSD17B1 is expressed in normal human endometrium, hyperplasia, and cancer. Collectively, our non-clinical data and literature analysis suggest that HSD17B1 inhibition could be one of several possible approaches to decrease endometrial estrogen production in endometrial hyperplasia and cancer. HSD17B1 expression has been found in bones of humans and rats. The non-clinical data in the present study suggest that human HSD17B1 is likely to have an important role in the regulation of bone formation, strength and length during reproductive years in female mice. Bone density in HSD17B1TG females was highly increased in femurs, but in lesser amounts also in tibias. Especially the tibia growth plate, but not other regions of bone, was susceptible to respond to HSD17B1 inhibition by increasing bone length, whereas the inhibitors did not affect bone density. Therefore, HSD17B1 inhibitors could be safer than aromatase inhibitors in regard to bone in the treatment of breast cancer and endometriosis. Furthermore, diseases related to improper growth, are a promising new indication for HSD17B1 inhibitors.

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Oral implantology is a common procedure in dentistry, especially for fully or partially edentulous patients. The implants must be placed in the best location from both the aesthetic and functional point of view. Because of this it is increasingly more frequent to resort to regeneration techniques that use substitutes of the bone itself, in order to be able to insert the implants in the most appropriate location. Material and Methodology: A review was performed on the literature from the last ten years based on the following search limitations: "graft materials', 'allograft', 'xenograft', 'autologous graft" and 'dentistry". Results: 241 works were obtained that after reading their respective summaries, they were reduced to 38, and 9 previous works were included in order to summarize the concepts. Discussion: Autologous grafts are the 'gold standard' of the bone regeneration. They have obvious advantages, but they also have drawbacks. This is why allogeneic and xenogeneic tissues are used. The former because of their clear similarity with the recipient's tissue and the latter due to their wide availability. Given that these grafts also have drawbacks, the industry has developed synthetic materials that have properties similar to those of human bone tissue. However, as of today, the ideal material to substitute human bone has not yet been found. In recent years the tendency has been to combine these synthetic materials with the patient's own bone, which is extracted during drilling in implant placement, with bone marrow aspiration, or with bone morphogenetic proteins. Thus the intention is to equip these substances with the osteogenic capacity. Conclusions: There is currently no ideal graft material, with the exception of those materials that come directly from the patient. We hope that in the coming years we will have products that will allow us to perform rehabilitations with better results and provide a better quality of life for our patients, especially those who have more complex situations to resolve, like the patients that are operated on for head and neck cancer

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A 40-year-old female patient presented abdominal pain during her periods and progressing constipation during the last 6 years. Retosigmoidoscopy showed a stenotic lesion 14cm above the anal verge. The patient underwent a retosigmoidectomy and the specimen was sent to histopathologic exam. The diagnose was endometriosis with the involvement of the colon and rectum, associated with dissemination of the endometrial cells by the lymph nodes. The patient recovered well and was discharged on the 6th postoperative day. It is commented on the rarity of the lymphatic dissemination in these cases and according to the reviewed literature, this is the 5th case reported. Thus, dissemination of the disease to extra-abdominal sites could possibly occur by this route. Therefore, pathologists should carefully exam the lymph nodes of the mesocolon in these specimens because findings of endometrial "metastasis" may be present.

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The practice of auto-transfusion is highly considered due to its safety and economic beneficts. It comprises two basic modalities: "Reinfusion" and "Pre-Collection" of blood. The reinfusion uses the patient's own blood spurted either from the patient's previous hemorrhage or during the surgery. The auto-transfusion of Pre-Collect uses the patient's blood tajen before surgery and it consists two basic modalities: The multiple Pre-Collection in which the blood is taken from the patient much before the scheduled sate of the surgery and the Pre-Collection done 10-30 minutes before surgery, which we presently designated as "Immediate Pre-Collection" (or Pre-Deposit for Immediate Utilization) which constitutes the basic topic of this essay.

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OBJETIVO: Desenvolver um modelo de endometriose experimental em ratas. MÉTODOS: Foram utilizadas 30 ratas adultas da linhagem Wistar. A técnica cirúrgica consistiu em laparotomia mediana com identificação do útero bicorno e ressecção de um segmento de 2 cm do corno uterino direito. Um retalho de 0,25 cm² foi retirado dessa estrutura e suturado na parede abdominal com a face endometrial voltada para a cavidade peritoneal. As ratas foram divididas aleatoriamente em dois grupos de acordo com o tempo para a reoperação: grupo 1 (n=15), reoperado em 30 dias, e grupo 2 (n=15), em 60 dias. No momento da segunda laparotomia os implantes foram avaliados macroscopicamente, ressecados e encaminhados para análise microscópica com coloração hematoxilina-eosina e imunohistoquímica (HEMA, AE1 e AE2). RESULTADOS: Os implantes se desenvolveram em 83,3 % do Grupo 1 e 71,4% no Grupo 2. Não houve diferença estatisticamente significativa entre o peso dos animals dos dois grupos. Também não houve diferença estatisticamente significativa no tamanho da área das lesões induzidas: no Grupo 1 a média foi 0,37 cm² e no Grupo 2, de 0,25 cm². Segundo a classificação histológica semi-quantitativa de Keenan (de acordo com a preservação da camada epitelial de endométrio), o Grupo 1 teve média de 1,9 e o Grupo 2, de 2,4. CONCLUSÃO: A técnica utilizada para o desenvolvimento de endometriose em ratas foi satisfatória.

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OBJETIVO: Avaliar o efeito da terapia anti-TNF-α no tratamento de implantes endometriais no peritônio de ratas. MÉTODOS: Os implantes endometrióticos foram induzidos cirurgicamente em 120 ratas Wistar-Albino. Os animais foram aleatoriamente distribuídos em 4 grupos. O grupo C (n=36) recebeu uma injeção intraperitoneal de 0,2ml de solução salina. O grupo L (n=41) recebeu uma injeção subcutânea de 1mg/kg de leuprolide. O grupo I5 (n=20) recebeu uma injeção subcutânea de 5mg/kg de anticorpo monoclonal anti-fator de necrose tumoral (TNF) a (infliximab). O grupo I10 (n=20) recebeu uma injeção subcutânea de 10mg/kg de infliximab. As ratas foram sacrificadas após 21 dias para se avaliar o tamanho dos implantes e a expressão do TNF-α. RESULTADOS: O tratamento com leuprolide promoveu uma redução absoluta na área de superfície do implante comparado com o grupo C (+14mm vs. 0mm; p=0,013) e com o grupo I10 (+14mm vs. +5mm; p=0,018). Da mesma forma, uma redução percentual da area de superfície do implante foi observada comparando o grupo L com o grupo C (+33,3% vs. 0%; p=0,005) e com o grupo I10 (+33,3% vs. +18,3%; p=0,027). O tratamento com infliximab não foi capaz de diminuir a área de superfície do implante comparado com o grupo C. A expressão de TNF-α reduziu nos grupos L, I5 e I10 comparado com o grupo C (505,6µm² vs. 660,5µm² vs. 317,2µm² vs. 2519,3µm², respectivamente; p<0,001). CONCLUSÃO: A terapia anti-TNF-α reduziu a expressão de TNF-α nos implantes endometrióticos mas não reduziu a área de superfície da lesão.

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OBJECTIVE: To assess hematological and biochemical features of splenic effluent blood and their influence on the rise of hematological values after splenectomy.METHODS: we studied 20 patients undergoing surgical treatment for schistosomatic portal hypertension. We collected blood samples for CBC, coagulation, bilirubin and albumin in the splenic vein (perioperative) and peripheral blood (immediately pre and postoperative periods).RESULTS: the splenic blood showed higher values of red blood cells, hemoglobin, hematocrit, platelet count, total leukocytes, neutrophils, lymphocytes, monocytes, eosinophils and basophils, as well as reduction of laboratory coagulation parameters in relation to peripheral blood collected preoperatively. In the postoperative peripheral blood there was an increase in the overall leukocytes and in their neutrophil component, and decreased levels of basophils, eosinophils and lymphocytes. The other postoperative variables of complete blood count and coagulation tests were not different compared with the splenic blood. The albumin values were lower postoperatively when compared to preoperative and splenic blood. There were higher values of direct bilirubin in the postoperative period when compared with the preoperative and splenic blood. Postoperative indirect bilirubin was lower compared to its value in the splenic blood.CONCLUSION: hematological and biochemical values of splenic effluent blood are higher than those found in peripheral blood in the presence of schistosomal splenomegaly. However, the splenic blood effluent is not sufficient to raise the blood levels found after splenectomy.

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The objective is to reinforce the importance of blood reinfusion as a cheap, safe and simple method, which can be used in small hospitals, especially those in which there is no blood bank. Moreover, even with the use of devices that perform the collection and filtration of blood, more recent studies show that the cost-benefit ratio is much better when autologous transfusion is compared with blood transfusions, even when there is injury to hollow viscera and blood contamination. It is known that the allogeneic blood transfusion carries a number of risks to patients, among them are the coagulation disorders mediated by excess enzymes in the conserved blood, and deficiency in clotting factors, mainly the Factor V, the proacelerin. Another factor would be the risk of contamination with still unknown pathogens or that are not investigated during screening for selection of donors, such as the West Nile Fever and Creutzfeldt-Jacob, better known as "Mad Cow" disease. Comparing both methods, we conclude that blood autotransfusion has numerous advantages over heterologous transfusion, even in large hospitals. We are not against blood transfusions, just do not agree that the patient's own blood is discarded without making sure there will be enough blood in stock to get him out of the hemorrhagic shock.

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The use of autologous platelet concentrates, represent a promising and innovator tools in the medicine and dentistry today. The goal is to accelerate hard and soft tissue healing. Among them, the platelet-rich plasma (PRP) is the main alternative for use in liquid form (injectable). These injectable form ofplatelet concentrates are often used in regenerative procedures and demonstrate good results. The aim of this study is to present an alternative to these platelet concentrates using the platelet-rich fibrin in liquid form (injectable) and its use with particulated bone graft materials in the polymerized form.

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Objetivo: determinar o papel da histerossonografia na avaliação das anormalidades da cavidade uterina, em pacientes com sangramento uterino anormal, selecionadas previamente pela ultra-sonografia transvaginal. Métodos: foram selecionadas 48 pacientes na menacme e pós-menopausa, portadoras de sangramento uterino anormal e anormalidades da cavidade uterina, diagnosticadas inicialmente pela ultra-sonografia transvaginal. Todas as pacientes se submeteram à histerossonografia e, posteriormente, aos métodos "padrão ouro", a histeroscopia e/ou histerectomia. As histerossonografias foram avaliadas por dois diferentes médicos e os diagnósticos confrontados. Resultados: a histerossonografia mostrou ter alta sensibilidade e especificidade no diagnóstico das patologias benignas da cavidade uterina. Na presença de pólipo a sensibilidade e a especificidade do método foram, respectivamente, de 100 e 97%, seguido do mioma submucoso cuja sensibilidade e especificidade foram, respectivamente, de 83 e 100%. Já na hiperplasia endometrial e no endométrio normal a sensibilidade e especificidade foram de 100%. Diagnosticamos 33 casos de pólipos, 13 casos de miomas submucosos, quatro casos de hiperplasia endometrial e três casos normais na análise dos métodos "padrão ouro". Houve alta correlação entre os diagnósticos dados pelos dois examinadores. Conclusões: a histerossonografia mostrou ser um exame reprodutível, com alta sensibilidade e especificidade no diagnóstico das patologias benignas da cavidade uterina em pacientes com sangramento uterino anormal.

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Objetivos: avaliar a eficácia do acetato de medroxiprogesterona e do acetato de megestrol nas hiperplasias de endométrio. Métodos: foram incluídas, retrospectivamente 47 pacientes com sangramento uterino anormal, submetidas a curetagem uterina diagnóstica e/ou biópsia de endométrio, cujo achado histopatológico foi de hiperplasia de endométrio. Nas pacientes com hiperplasia sem atipia foi iniciado a terapêutica com acetato de medroxiprogesterona por via oral, na dose de 10 mg/dia durante 10-12 dias por mês. Nas com atipia, era utilizado o acetato de megestrol por via oral, dose de 160 mg/dia, uso contínuo. O período de tratamento variou de 3 a 18 meses. Biópsia de endométrio e/ou curetagem uterina de controle foram realizadas entre três e seis meses do início do tratamento e periodicamente para avaliar a resposta terapêutica. Resultados: foram analisadas 42 pacientes com hiperplasia endometrial sem atipia e cinco com atipia. A média de idade das pacientes foi de 49,5 ± 10,6 anos, sendo 70,2% com idade superior a 45 anos. O acetato de medroxiprogesterona foi eficaz em fazer regredir as hiperplasias sem atipias em 83,2% (35/42) e o acetato de megestrol em 80% (4/5) das hiperplasias com atipia. Em 16,8% (7 casos) das hiperplasias sem atipia e em 20% (1 caso) das com atipia, ocorreu persistência das lesões, apesar do tratamento. Em nenhum caso ocorreu progressão para câncer de endométrio, durante o período de seguimento que foi de 3 meses a 9 anos. No acompanhamento dessas pacientes, verificamos que 18 (38,3%) apresentaram amenorréia, em 12 (25,5%) ocorreu regularização do ciclo menstrual e 17 (36,2%) permaneceram com sangramento uterino anormal, sendo submetidas a histerectomia total abdominal. O exame anatomopatológico mostrou a persistência da lesão hiperplásica em oito casos, leiomioma em quatro, adenomiose em três, mio-hipertrofia uterina difusa em um caso e útero normal em outro, tendo havido regressão das lesões hiperplásicas nesses últimos nove casos. Conclusões: o tratamento das hiperplasias de endométrio com acetato de medroxiprogesterona e/ou acetato de megestrol, representa uma alternativa satisfatória para mulheres que desejam preservar o útero ou que tenham risco cirúrgico elevado. Entretanto, é necessário monitorização cuidadosa do endométrio, o que deve ser realizado pela avaliação dos sintomas, ultra-sonografia transvaginal e biópsia periódica.

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Objetivo: avaliar criticamente os achados histopatológicos e a relação custo/benefício da dilatação e curetagem uterina (D&C) no rastreio do sangramento uterino anormal (SUA). Método: análise retrospectiva dos resultados histopatológicos de 542 D&C praticadas por SUA na Disciplina de Ginecologia da Faculdade de Ciências Médicas da Universidade do Estado do Rio de Janeiro (FCM-UERJ), de janeiro de 1984 a janeiro de 1994. As pacientes foram divididas em dois grupos: Grupo 1 - pacientes com idade igual ou inferior a 50 anos (n = 385) e Grupo 2 - pacientes com mais de 50 anos (n = 157). Foram excluídos desse estudo os casos de curetagens de urgência. Todas as curetagens foram realizadas sob narcose. O tempo médio de internação foi de três dias. Considerou-se resultado patológico negativo quando o laudo histopatológico mostrou endométrio tipo proliferativo, secretor, atrófico ou iatrogênico. Este último termo refere-se a endométrio sob possível influência de medicação hormonal. Considerou-se resultado patológico positivo quando o laudo histopatológico evidenciou algum tipo de lesão. Resultados: no Grupo 1 encontrou-se resultado patológico negativo em 50,2% dos casos, resultado patológico positivo em 39,7% dos casos e material insuficiente para diagnóstico (MIPD) em 10,1% dos casos. Pólipo endometrial e mioma submucoso foram observados em apenas 5,5% e 4,4%, respectivamente. O câncer foi de observação incomum nesse grupo, sendo encontrado o adenocarcinoma do endométrio (ACE) em apenas 1,3% dos casos (n = 5), numa relação de 77 D&C para um ACE. No Grupo 2 observou-se resultado patológico negativo em 38,3% dos casos, resultado patológico positivo em 38,1% dos casos e MIPD em 23,6% dos casos. Pólipo endometrial e mioma submucoso foram diagnosticados em somente 5,1% e 0,6%, respectivamente. Lesões malignas foram encontradas em 12% dos casos, sendo 9,5% (15 casos) de ACE, mostrando relação de um ACE para 10 D&C. Conclusões: consoante o conhecimento atual sobre a etiopatogenia do SUA este estudo mostrou que a D&C diagnóstica tradicional tem baixa acurácia na avaliação daquele sangramento e relação custo/benefício incompatível com a medicina atual. Portanto, não deve ser o exame de primeira escolha. Considerando, contudo, que o ACE foi encontrado em uma de cada 10 D&C em mulheres com mais de 50 anos com queixa de sangramento uterino, pode-se indicar D&C com mais liberalidade nesse grupo, uma vez que não se disponha de histeroscopia com biópsia dirigida. Atualmente, a D&C não tem mais um papel significante no rastreio do SUA como tinha há alguns anos. Entretanto, o procedimento ainda encontra indicação em algumas situações e não pode ser de todo abandonado, devendo sua indicação obedecer a critérios restritos.