Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 10 de 10
Filtrar
1.
Minerva Obstet Gynecol ; 74(6): 504-515, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34851073

RESUMO

BACKGROUND: Intrauterine adhesions caused by postpartum curettage, spontaneous abortions, interrupted pregnancies, endometrial ablations, infections and inflammations, can lead to a loss of endometrial function, with consequent hypomenorrhea and infertility in women of reproductive age. In a non-negligible percentage of cases, the available surgical methods and hormone therapy, with sequential administration of estrogen and progesterone, are ineffective. In fact, severe damage to the basal layer of the endometrium causes the loss of endometrial cell precursors and leads to the failure of regeneration of the functional layer to which the endometrium is cyclically exposed. Today, many researchers are evaluating the use of stem cells of different origins as a potential therapy to restore endometrial function. METHODS: Our interest has been focused on adipose-derived stromal/stem cells (ADSCs) obtained by collecting subcutaneous adipose tissue and subsequently treating it with the MilliGraft® method. This procedure produces a cell suspension, the stromal vascular fraction (SVF), which includes ADSCs and soluble factors such as proteins and extracellular vesicles (exosomes). The SVF thus obtained was characterized in its cellular composition and its functional factors. Our clinical protocol for the future use of adipose tissue in endometrial regeneration in its different phases is presented. RESULTS: The data obtained, even though they still require further support and implementation, show the regenerative properties of SVF obtained from adipose tissue using a mechanical method. CONCLUSIONS: These findings can contribute to the development of cell therapies using stem cells of different derivations which are increasingly being utilized in the treatment of endometrial lesions from adherent or dysfunctional pathologies.


Assuntos
Endométrio , Doenças Uterinas , Gravidez , Feminino , Humanos , Endométrio/metabolismo , Transplante de Células-Tronco/métodos , Tecido Adiposo/metabolismo , Células Estromais/metabolismo , Doenças Uterinas/metabolismo , Aderências Teciduais/metabolismo
2.
Int J Mol Sci ; 22(4)2021 Feb 19.
Artigo em Inglês | MEDLINE | ID: mdl-33669517

RESUMO

Despite low levels of vascular endothelial growth factor (VEGF)-A, the secretome of human Wharton's jelly (WJ) mesenchymal stromal cells (MSCs) effectively promoted proangiogenic responses in vitro, which were impaired upon the depletion of small (~140 nm) extracellular vesicles (EVs). The isolated EVs shared the low VEGF-A profile of the secretome and expressed five microRNAs, which were upregulated compared to fetal dermal MSC-derived EVs. These upregulated microRNAs exclusively targeted the VEGF-A gene within 54 Gene Ontology (GO) biological processes, 18 of which are associated with angiogenesis. Moreover, 15 microRNAs of WJ-MSC-derived EVs were highly expressed (Ct value ≤ 26) and exclusively targeted the thrombospondin 1 (THBS1) gene within 75 GO biological processes, 30 of which are associated with the regulation of tissue repair. The relationship between predicted microRNA target genes and WJ-MSC-derived EVs was shown by treating human umbilical-vein endothelial cells (HUVECs) with appropriate doses of EVs. The exposure of HUVECs to EVs for 72 h significantly enhanced the release of VEGF-A and THBS1 protein expression compared to untreated control cells. Finally, WJ-MSC-derived EVs stimulated in vitro tube formation along with the migration and proliferation of HUVECs. Our findings can contribute to a better understanding of the molecular mechanisms underlying the proangiogenic responses induced by human umbilical cord-derived MSCs, suggesting a key regulatory role for microRNAs delivered by EVs.


Assuntos
Vesículas Extracelulares/metabolismo , Células-Tronco Mesenquimais/metabolismo , MicroRNAs/metabolismo , Neovascularização Fisiológica , Fator A de Crescimento do Endotélio Vascular/metabolismo , Geleia de Wharton/citologia , Movimento Celular , Proliferação de Células , Separação Celular , Feto/citologia , Fluoresceínas/metabolismo , Ontologia Genética , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Imunofenotipagem , MicroRNAs/genética , Nanopartículas/química , Reprodutibilidade dos Testes , Pele/citologia , Succinimidas/metabolismo , Trombospondina 1/metabolismo , Cordão Umbilical/citologia
3.
J Assist Reprod Genet ; 30(4): 497-503, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23435529

RESUMO

PURPOSE: An observational clinical and molecular study was designed to evaluate the effects of the administration of recombinant human FSH on sperm DNA fragmentation in men with a non-classical form of hypogonadotropic hypogonadism and idiopathic oligoasthenoteratozoospermia. METHODS: In the study were included 53 men with a non-classical form of hypogonadotropic hypogonadism and idiopathic oligoasthenoteratozoospermia. In all patients, sperm DNA fragmentation index (DFI), assessed by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate (dUTP) in situ DNA nick end-labelling (TUNEL) assay, was evaluated before starting the treatment with 150 IU of recombinant human FSH, given three times a week for at least 3 months. Patients' semen analysis and DNA fragmentation index were re-evaluated after the 3-month treatment period. RESULTS: After recombinant human FSH therapy, we did not find any differences in terms of sperm count, motility and morphology. The average DNA fragmentation index was significantly reduced (21.15 vs 15.2, p<0.05), but we found a significant reduction in patients with high basal DFI values (>15 %), while no significant variation occurred in the patients with DFI values ≤ 15 %. CONCLUSIONS: Recombinant human FSH administration improves sperm DNA integrity in hypogonadotropic hypogonadism and idiopathic oligoasthenoteratozoospermia men with DNA fragmentation index value >15 % .


Assuntos
Fragmentação do DNA , Hormônio Foliculoestimulante Humano/administração & dosagem , Hipogonadismo/metabolismo , Espermatozoides/metabolismo , Adulto , Astenozoospermia/genética , Astenozoospermia/metabolismo , Humanos , Hipogonadismo/genética , Masculino , Oligospermia/genética , Oligospermia/metabolismo , Proteínas Recombinantes/administração & dosagem , Contagem de Espermatozoides , Espermatozoides/citologia
4.
J Assist Reprod Genet ; 29(11): 1281-7, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22968514

RESUMO

OBJECTIVE: To evaluate the relationship between sperm nuclear vacuoles and sperm morphology and to investigate the influence of the rate of spermatozoa with head vacuolization (SVR) in a seminal sample on the clinical outcomes in couples undergoing intracytoplasmic sperm injection. MATERIALS: 26 patients undergoing infertility investigations were included and were divided in two groups according to an SVR ≤ 20,28 % (Group A) or > 20,28 % (Group B), and were investigated to verify the influence of SVR on the fertilization rate, embryo quality, pregnancy and implantation rates. RESULTS: Abnormal spermatozoa with nuclear vacuoles were significantly higher (p < 0.001) than the percentage of normal spermatozoa with nuclear vacuoles. Patients in group A had a percentage of abnormal sperm with nuclear vacuole significantly lower compared to group B (p < 0,001), but there was no difference in the percentage of normal sperm with nuclear vacuoles. Fertilization rates and the number of top quality embryos did not differ between the two groups. The pregnancy and implantation rates were significantly higher in Group A compared to Group B (respectively p < 0,05 and p < 0.001). CONCLUSIONS: For the first time, we propose a cut off value in the proportion of sperms with nuclear vacuolization on the total of sperm in seminal samples, and demonstrate a relationship between SNV and clinical outcomes after ICSI. The SNV rate could be introduced as an easy diagnostic evaluation prior to perform an ICSI cycle.


Assuntos
Núcleo Celular/ultraestrutura , Cabeça do Espermatozoide/ultraestrutura , Injeções de Esperma Intracitoplásmicas/normas , Espermatozoides/anormalidades , Adulto , Feminino , Humanos , Masculino , Gravidez , Técnicas de Reprodução Assistida/normas , Injeções de Esperma Intracitoplásmicas/métodos , Espermatozoides/citologia , Espermatozoides/ultraestrutura
5.
Fertil Steril ; 96(6): 1384-1390.e4, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21982727

RESUMO

OBJECTIVE: To test whether metformin administration reduces the incidence of ovarian hyperstimulation syndrome (OHSS) in infertile high-risk patients with polycystic ovary syndrome (PCOS) who have been treated with gonadotropins for IVF. DESIGN: Parallel, randomized, double-blind, placebo-controlled clinical trial. SETTING: Academic departments, general hospital, and IVF centers. PATIENT(S): One hundred twenty patients with PCOS at high risk for OHSS. INTERVENTION(S): Gonadotropins ovarian stimulation for IVF and metformin (500 mg three times daily) or placebo tablets (three times daily). MAIN OUTCOME MEASURE(S): The primary end point of the current clinical trial was the rate of OHSS. Anthropometric and reproductive data were evaluated. RESULT(S): The total OHSS and cancellation rates were significantly reduced in patients treated with metformin. The relative risk for OHSS was of 0.28 (95% confidence interval, 0.11-0.67). With metformin the stimulation length and the total amount of gonadotropins used were significantly increased, whereas the peak E(2) levels were significantly reduced. CONCLUSION(S): In patients with PCOS who are at high risk for OHSS and who have been stimulated with gonadotropins for IVF cycles, metformin reduces the risk of OHSS by modulating the ovarian response to the stimulation. REGISTRATION ID NUMBER FROM CLINICALTRIALS.GOV: NCT01233206.


Assuntos
Fertilização in vitro , Gonadotropinas/uso terapêutico , Metformina/uso terapêutico , Síndrome de Hiperestimulação Ovariana/prevenção & controle , Síndrome do Ovário Policístico/tratamento farmacológico , Administração Oral , Adulto , Método Duplo-Cego , Regulação para Baixo/efeitos dos fármacos , Esquema de Medicação , Feminino , Fertilização in vitro/métodos , Humanos , Hipoglicemiantes/administração & dosagem , Hipoglicemiantes/farmacologia , Hipoglicemiantes/uso terapêutico , Metformina/administração & dosagem , Metformina/farmacologia , Síndrome de Hiperestimulação Ovariana/epidemiologia , Síndrome de Hiperestimulação Ovariana/etiologia , Indução da Ovulação/métodos , Placebos , Síndrome do Ovário Policístico/complicações , Gravidez , Fatores de Risco
7.
Fertil Steril ; 87(3): 542-6, 2007 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-17126339

RESUMO

OBJECTIVE: To investigate the effects of recombinant (r-) LH supplementation in "low responder" patients undergoing ovarian stimulation with r-FSH for an IVF program. The apoptosis rate in cumulus cells was used as an indicator of oocyte quality. DESIGN: Comparison of the rate of DNA fragmentation and caspase-3 activity in cumulus cells in women stimulated with r-LH and r-FSH, versus patients treated with r-FSH alone (control). SETTING: In vitro fertilization (IVF) laboratory. PATIENT(S): Forty patients undergoing assisted fertilization programs treated with a GnRH agonist, or r-FSH treatment begun on day 3 of the cycle (control). In the r-LH group, from day 8 of gonadotropin stimulation, 150 IU per day of r-LH were administered. INTERVENTION(S): Terminal deoxynucleotidyl transferase-mediated digoxigenin-deoxyuridine-triphosphate (dUTP) nick-end labeling (TUNEL) assay, and anti-caspase-3 cleaved immunoassay, to detect apoptosis in human cumulus cells. MAIN OUTCOME MEASURE(S): Difference in DNA fragmentation rate between cumulus cells derived from r-LH treatment and cumulus cells derived from control patients. RESULT(S): No differences were observed between the two groups in the total amount of r-FSH administered and in the number of retrieved oocytes per patient. A statistically significant increase in the number of immature oocytes and in the E(2) serum peak was observed in the control group. The number of transferred embryos was significantly higher in the r-LH group. Pregnancy and implantation rates were higher in the r-LH group, but without statistical significance. The apoptosis rate in cumulus cells was higher in the control group than in the r-LH group. CONCLUSION(S): This study suggests that supplementation with r-LH improves the chromatin quality of cumulus cells involved in the control of oocyte maturation.


Assuntos
Apoptose , Células da Granulosa/citologia , Hormônio Luteinizante/uso terapêutico , Indução da Ovulação/métodos , Adulto , Caspase 3/análise , Fragmentação do DNA , Feminino , Fertilização in vitro , Hormônio Foliculoestimulante/uso terapêutico , Humanos , Marcação In Situ das Extremidades Cortadas , Gravidez , Taxa de Gravidez , Proteínas Recombinantes/uso terapêutico
9.
Fertil Steril ; 84(5): 1417-23, 2005 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-16275238

RESUMO

OBJECTIVE: To investigate the presence of programmed cell death in unfertilized oocytes after intracytoplasmic sperm injection (ICSI), assuming that previous apoptotic events could be correlated with the fertilization failure. DESIGN: Comparison of the rate of DNA fragmentation in human oocytes at different stages of maturation soon after pick-up (control) and in unfertilized oocytes after ICSI treatment. SETTING: In vitro fertilization (IVF) laboratory with extensive ICSI experience. PATIENT(S): Sixty-three patients undergoing assisted fertilization by ICSI. INTERVENTION(S): Terminal deoxynucleotidyl transferase-mediated digoxigenin-dUTP nick-end labeling (TUNEL) assay and anticaspase-3 cleaved immunoassay to detect apoptosis in control and ICSI-treated oocytes. MAIN OUTCOME MEASURE(S): Differences in the percentage of oocytes demonstrating DNA fragmentation between control oocytes and unfertilized ICSI treated oocytes at different stages of maturation. RESULT(S): The DNA fragmentation, by TUNEL assay, appeared in all the immature control oocytes, but only 37% of mature oocytes showed DNA fragmentation. This DNA fragmentation was observed in 88.8% of the oocytes unfertilized after ICSI; furthermore, DNA fragmentation appeared as well in the sperm injected into the cytoplasm. CONCLUSION(S): The study has shown DNA fragmentation in human oocytes unfertilized after ICSI. The evidence is confirmed as well in control oocytes, free from in vitro culture or manipulation stress. Caspase-3 immunoassay suggests the presence of apoptosis. The high percentage of oocytes demonstrating DNA fragmentation in the unfertilized oocytes could be correlated with fertilization failure.


Assuntos
Apoptose/fisiologia , Fragmentação do DNA/fisiologia , Oócitos/citologia , Oócitos/fisiologia , Injeções de Esperma Intracitoplásmicas/métodos , Distribuição de Qui-Quadrado , Intervalos de Confiança , Feminino , Humanos
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...