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1.
Ann Transl Med ; 12(2): 29, 2024 Apr 22.
Artículo en Inglés | MEDLINE | ID: mdl-38721455

RESUMEN

Simulated operations (SOs) are a direct application of the Integral Theory (IT) mantras, "structure and function are related" and "restore the structure and you will improve the function". SOs performed in a clinic setting, are the most effective way possible to test the validity of the IT predictions: stress urinary incontinence (SUI) and urge are mainly caused by laxity in the vagina or its supporting ligaments. The SUI prediction of the IT is validated if a hemostat applied vaginally in the position of the midurethra to mechanically support the pubourethral ligament (PUL) immediately stops urine loss on coughing. The urge and chronic pelvic pain (CPP) predictions of the IT are similarly validated if a patient states her urge and pain symptoms are relieved by insertion of the bottom blade of a bivalve speculum which supports the uterosacral ligaments (USLs). An important use of SOs is to preoperatively assess (by the hemostat test) whether sling surgery for SUI is likely to cure the patient. Similarly, the speculum is very useful for diagnosing whether severe urge or pain symptoms in a woman with minimal prolapse are originating from weak USLs. If digital support of a cystocele relieves urge symptoms, the patient can reasonably be informed that a cystocele repair should improve the urge as well her cystocele prolapse. Used intraoperatively under spinal anesthesia, SOs can determine whether a sling is sufficiently tight to reverse the loose PUL which is causing the SUI. Approximating both cardinal ligaments (CLs) intraoperatively can result in a remarkable disappearance of a transverese defect cystocele; approximating USLs intraoperatively can give an indication of how effective a USL plication would be surgically.

4.
Stem Cell Res Ther ; 13(1): 331, 2022 07 23.
Artículo en Inglés | MEDLINE | ID: mdl-35870968

RESUMEN

BACKGROUND: Due to the large area and deep width of the artificial neovagina after vaginoplasty, it takes a considerable amount of time to achieve complete epithelization of the neovagina. Currently, the clinical therapies for vaginal epithelization after vaginoplasty are still dissatisfactory. Recent studies showed that small extracellular vesicles (sEVs) derived from stem cells could accelerate wound epithelization. The sustained release of sEVs from optimized hydrogels may be a promising strategy to accelerate vaginal epithelization after vaginoplasty. METHODS: The efficacy of phototriggered imine crosslink hydrogels (piGEL) containing sEVs derived from human urine-derived stem cells (hUSC-sEVs, piGEL-sEVs) on vaginal mucosa defects in rabbits was assessed by wound closure rates, histological analysis and immunofluorescence staining analysis. Cell counting kit-8, 5-ethynyl-2'-deoxyuridine and scratch wound assays were performed to assess the effects of hUSC-sEVs on the proliferation and migration ability of vaginal epithelial cells (VK2/E6E7). Quantitative real-time polymerase chain reaction (qRT-PCR) was carried out to test the expression of epithelial differentiation markers in VK2 cells. Moreover, a microRNA (miRNA) microarray was used to find hUSC-sEVs-specific miRNAs that potentially affected the proliferation, migration and differentiation ability of VK2 cells. RESULTS: The in vitro release profile revealed that the piGEL could ensure sustained release of hUSC-sEVs. The in vivo results showed that piGEL-sEVs effectively promoted epithelization and angiogenesis of vaginal mucosa defects in rabbits. According to miRNA microarray and qRT-PCR results, miR-126-3p might be the crucial molecule among the various miRNAs contained in hUSC-sEVs. The data showed that hUSC-sEVs promoted the migration and differentiation of VK2 cells by delivering miR-126-3p to suppress the expression of Spred1 and PIK3R2, thereby activating the ERK1/2 and ATK signaling pathways. CONCLUSION: The results indicated that piGEL-sEVs could be a novel promising approach for enhancing the epithelization of the neovagina after vaginoplasty and provided useful data for understanding the underlying mechanism of the effect of hUSC-sEVs on epithelization.


Asunto(s)
Vesículas Extracelulares , MicroARNs , Animales , Preparaciones de Acción Retardada/metabolismo , Vesículas Extracelulares/metabolismo , Femenino , Humanos , Hidrogeles/farmacología , Iminas/metabolismo , MicroARNs/genética , MicroARNs/metabolismo , Conejos , Células Madre/metabolismo
5.
Mol Cell Biochem ; 477(5): 1629-1643, 2022 May.
Artículo en Inglés | MEDLINE | ID: mdl-35229243

RESUMEN

Precise differentiation of glucokinase (GCK) monogenic diabetes from gestational diabetes mellitus (GDM) is critical for accurate management of the pregnancy outcome. We screened GCK-MODY complicating pregnancies in Chinese GDM patients, explored the pathogenesis of novel GCK mutations, and evaluated the patients' pregnancy outcome and management. The GCK gene from 411 GDM patients was screened with PCR-direct sequencing and multiplex ligation-dependent probe amplification (MLPA) and 15 GCK mutations were identified. We also retrospectively analyzed a total of 65 pregnancies from 21 GCK-MODY families, wherein 41 were from 15 maternal families and 24 were from six paternal families. Bioinformatic analysis and biochemical functional study were conducted to identify novel GCK mutations. In total, we identified 21 GCK mutations: 15 from the 411 GDM patients and six from 24 fathers. Of th Asp78Asn (GAC → AAC), Met87Arg (ATG → AGG), Leu451Val (CTT → GTT), Leu451Pro (CTG → CCG) and 1019 + 20G > A e mutations, five, i.e., were novel and deleterious, with markedly decreased enzyme activity and thermal stability. The unaffected offspring of GCK mutation-affected mothers were heavier than affected offspring (p < 0.001). Of 21 insulin-treated affected mothers, 10 had maternal hypoglycemia (47.6%) and seven had perinatal complications (33.3%), and the affected offspring of the insulin-treated affected mothers had significantly lower birth weights than that of the 20 diet-control affected mothers (p = 0.031). In this study, the prevalence of GCK-MODY complicating pregnancy in Chinese GDM patients was 3.6% (15/411). The defective GCK may contribute to the hyperglycemia in GCK-MODY. Insulin therapy is not beneficial for GCK-MODY complicating pregnancy and therefore should not be recommended.


Asunto(s)
Diabetes Mellitus Tipo 2 , Diabetes Gestacional , Embarazo en Diabéticas , China , Diabetes Mellitus Tipo 2/genética , Diabetes Gestacional/genética , Femenino , Glucoquinasa/genética , Humanos , Insulina/genética , Mutación , Embarazo , Resultado del Embarazo , Embarazo en Diabéticas/epidemiología , Embarazo en Diabéticas/genética , Embarazo en Diabéticas/terapia , Estudios Retrospectivos
6.
Chin Med J (Engl) ; 134(2): 200-205, 2021 Jan 20.
Artículo en Inglés | MEDLINE | ID: mdl-33443938

RESUMEN

BACKGROUND: It has been a global trend that increasing complications related to pelvic floor surgeries have been reported over time. The current study aimed to outline the development of Chinese pelvic floor surgeries related to pelvic organ prolapse (POP) over the past 14 years and investigate the potential influence of enhanced monitoring conducted by the Chinese Association of Urogynecology since 2011. METHODS: A total of 44,594 women with POP who underwent pelvic floor surgeries between October 1, 2004 and September 30, 2018 were included from 22 tertiary academic medical centers. The data were reported voluntarily and obtained from a database. We compared the proportion of each procedure in the 7 years before and 7 years after September 30, 2011. The data were analyzed by performing Z test (one-sided). RESULTS: The number of different procedures during October 1, 2011-September 30, 2018 was more than twice that during October 1, 2004-September 30, 2011. Regarding pelvic floor surgeries related to POP, the rate of synthetic mesh procedures increased from 38.1% (5298/13,906) during October 1, 2004-September 30, 2011 to 46.0% (14,107/30,688) during October 1, 2011-September 30, 2018, whereas the rate of non-mesh procedures decreased from 61.9% (8608/13,906) to 54.0% (16,581/30,688) (Z = 15.53, P < 0.001). Regarding synthetic mesh surgeries related to POP, the rates of transvaginal placement of surgical mesh (TVM) procedures decreased from 94.1% (4983/5298) to 82.2% (11,603/14,107) (Z = 20.79, P < 0.001), but the rate of laparoscopic sacrocolpopexy (LSC) procedures increased from 5.9% (315/5298) to 17.8% (2504/14,107). CONCLUSIONS: The rate of synthetic mesh procedures increased while that of non-mesh procedures decreased significantly. The rate of TVM procedures decreased while the rate of LSC procedures increased significantly. TRIAL REGISTRATION NUMBER: NCT03620565, https://register.clinicaltrials.gov.


Asunto(s)
Diafragma Pélvico , Prolapso de Órgano Pélvico , China , Femenino , Procedimientos Quirúrgicos Ginecológicos/efectos adversos , Humanos , Diafragma Pélvico/cirugía , Prolapso de Órgano Pélvico/cirugía , Mallas Quirúrgicas/efectos adversos , Resultado del Tratamiento , Vagina
7.
Stem Cell Res Ther ; 11(1): 3, 2020 01 03.
Artículo en Inglés | MEDLINE | ID: mdl-31900201

RESUMEN

BACKGROUND: Premature ovarian failure (POF) has a great impact on reproductive endocrine function in females, and it is an important cause of infertility. Previous studies have demonstrated that small extracellular vesicles (sEVs) derived from stem cells play an important role in tissue regeneration. This study aimed to investigate the therapeutic effect of sEVs derived from embryonic stem cells (ESCs-sEVs) on damaged ovaries and explore the underlying molecular mechanisms. METHODS: Mice POF models were established by injecting mice with cyclophosphamide and busulfan. Then, ESCs-sEVs were intravenously transplanted into POF mice. The plasma of mice was harvested at 1 and 2 weeks after treatment to analyze the levels of anti-Mullerian hormone (AMH), estradiol (E2), and follicle stimulating hormone (FSH) by ELISA. The morphology of ovaries and follicles was observed by H&E staining, and apoptosis of granulosa cells was detected by TUNEL. In vitro, EdU and CCK-8 tests were used to evaluate the proliferation of cultured granulosa cells stimulated by ESCs-sEVs. Western blotting was used to determine the expression of PI3K/AKT and apoptotic-related proteins. RESULTS: After transplantation of ESCs-sEVs, the levels of serum sex hormones recovered to normal levels. In addition, the number of follicles was significantly increased, and the number of apoptotic cells was decreased. The results in vitro revealed that ESCs-sEVs could significantly improve the proliferation rate of granulosa cells and increase the expression of phosphorylated PI3K and AKT. Meanwhile, the positive effect on proliferation and the negative effect on apoptosis observed in granulosa cells were obviously decreased when the PI3K/AKT signaling pathway was inhibited. CONCLUSION: Our findings suggested that ESCs-sEVs could improve ovarian function by regulating the PI3K/AKT signaling pathway, which could provide a promising clinical therapy for POF.


Asunto(s)
Células Madre Embrionarias/metabolismo , Vesículas Extracelulares/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Insuficiencia Ovárica Primaria/terapia , Proteínas Proto-Oncogénicas c-akt/metabolismo , Animales , Femenino , Humanos , Ratones , Transducción de Señal
8.
Biochem Biophys Res Commun ; 516(4): 1167-1174, 2019 09 03.
Artículo en Inglés | MEDLINE | ID: mdl-31284954

RESUMEN

Promotion of wound healing is one of the most important fields in clinical medical research. This study aimed to evaluate the potential use of a new surface-structured bacterial cellulose(S-BC) biomaterial with human urine-derived stem cells (hUSCs) for wound healing. In vitro, EA.hy926 were inoculated on structured/non-structured bacterial cellulose, and the growth of EA.hy926 on bacterial cellulose in medium with/without conditioned medium of the hUSCs were observed to explore the effect of bacterial cellulose's surface structure and hUSCs-CM on vascular endothelial cell growth. In vivo, we covered wound surface with various BC materials and/or injected the hUSCs into the wound site on group BC, group S-BC, group hUSCs, group BC + hUSCs, group S-BC + hUSCs to evaluate the effect of S-BC and hUSCs on wound healing in rat full-thickness skin defect model. In vitro study, surface structure of S-BC could promote the growth and survival of EA.hy926, and the hUSCs-CM could further promote the proliferation of EA.hy926 on S-BC. In vivo study, wound healing rate of the group BC, group S-BC, group hUSCs was significantly accelerated, accompanied by faster re-epithelialization, collagen production and neovascularization than control group. It is note worthy that the effect of S-BC on wound healing was better than BC, the effect of S-BC + hUSCs on wound healing was better than BC + hUSCs. Moreover, the effect of S-BC combined with hUSCs on wound is better than treated with S-BC or hUSCs alone. All the findings suggest that the combination of S-BC and hUSCs could facilitate skin wound healing by promoting angiogenesis. This combination of the role of stem cells and biomaterial surface structures may provide a new way to address clinical wound healing problems.


Asunto(s)
Materiales Biocompatibles/uso terapéutico , Celulosa/uso terapéutico , Neovascularización Fisiológica , Trasplante de Células Madre , Cicatrización de Heridas , Animales , Materiales Biocompatibles/química , Línea Celular , Células Cultivadas , Celulosa/química , Células Endoteliales/citología , Humanos , Polisacáridos Bacterianos/química , Polisacáridos Bacterianos/uso terapéutico , Ratas , Trasplante de Células Madre/métodos , Células Madre/citología , Propiedades de Superficie , Andamios del Tejido/química
9.
Stem Cell Res Ther ; 10(1): 80, 2019 03 08.
Artículo en Inglés | MEDLINE | ID: mdl-30849996

RESUMEN

BACKGROUND: Previous studies revealed that urine-derived stem cells (USCs) could promote myogenesis after the impairment of the sphincter muscles. However, the effects of exosomes secreted by USCs (USCs-Exo) were not elucidated. Exosomes are nanosized membrane vesicles secreted by the cells. They have been proved to be effective in protecting against tissue injury and therapeutic in tissue repair. USCs are ideal sources of exosomes because of the noninvasive obtaining method and self-renewal abilitiy. This study aimed to show the therapeutic effects of USCs-Exo on improving stress urinary incontinence (SUI). METHODS: Rat SUI models were established in this study using vaginal balloon inflation, and urodynamic and histological examination were carried out after exosome application. The proliferation and differentiation of muscle satellite cells (SCs) were evaluated using EdU, Cell Counting Kit 8, immunofluorescence staining, and Western blot analysis. mRNAs and proteins related to the activation of SCs were detected by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) and Western blot analysis. RESULTS: After exosome injection, the urodynamic parameters significantly improved and the injured muscle tissue recovered well. The activation, proliferation, and differentiation of SCs were promoted. The phosphorylation of extracellular-regulated protein kinases (ERK) was enhanced. When ERK was inhibited, the promoting effect of USCs-Exo treatment disappeared. CONCLUSION: The findings of this study elucidated the functional roles of USCs-Exo in satellite cell ERK phosphorylation and identified a novel agent for skeletal muscle regeneration, providing a basis for further exploring a cell-free correction for SUI.


Asunto(s)
Diferenciación Celular , Exosomas , Desarrollo de Músculos , Músculo Liso , Células Satélite del Músculo Esquelético , Incontinencia Urinaria de Esfuerzo , Orina , Animales , Exosomas/metabolismo , Exosomas/patología , Exosomas/trasplante , Femenino , Humanos , Músculo Liso/lesiones , Músculo Liso/metabolismo , Músculo Liso/patología , Ratas , Ratas Sprague-Dawley , Células Satélite del Músculo Esquelético/metabolismo , Células Satélite del Músculo Esquelético/patología , Incontinencia Urinaria de Esfuerzo/metabolismo , Incontinencia Urinaria de Esfuerzo/patología , Incontinencia Urinaria de Esfuerzo/terapia
10.
Immunology ; 156(3): 235-248, 2019 03.
Artículo en Inglés | MEDLINE | ID: mdl-30418673

RESUMEN

This study focuses on investigating the role of interleukin-1ß (IL-1ß) in functional regeneration following nerve injury in mice. A microarray-based mRNA profiling study was used to analyze the expression level of IL-1ß in peripheral nerve regeneration. Quantitative real-time polymerase chain reaction and Western blot were applied to assess the IL-1ß expressions of C57BL/6J-crush and C57BL/6J-crush+IL-1ß mice at different post-injury time-points after the standard sciatic nerve crush injury. The outcomes of nerve regeneration were evaluated by behavioral tests. IL-1ß was found to be up-regulated in peripheral nerve regeneration and significantly raised on the 3rd day and returned to normal levels on the 14th day after nerve injury. Compared with C57BL/6J-crush+IL-1ß mice, the nerve regeneration of C57BL/6J-crush mice was worse after nerve crush injury. IL-1ß increased mechanical sensitivity and stimulated amplitude. IL-1ß could benefit the recovery of sciatic nerve crush injury by facilitating nerve regeneration.


Asunto(s)
Interleucina-1beta/inmunología , FN-kappa B/inmunología , Regeneración Nerviosa/inmunología , Enfermedades del Sistema Nervioso/inmunología , Transducción de Señal/inmunología , Animales , Modelos Animales de Enfermedad , Femenino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Regulación hacia Arriba/inmunología
11.
Stem Cell Res Ther ; 9(1): 348, 2018 12 13.
Artículo en Inglés | MEDLINE | ID: mdl-30545407

RESUMEN

BACKGROUND: This study was aimed to investigate the role and specific molecular mechanism of HIF1A-AS2/miR-665/IL6 axis in regulating osteogenic differentiation of adipose-derived stem cells (ASCs) via the PI3K/Akt signaling pathway. METHODS: RNAs' expression profile in normal/osteogenic differentiation-induced ASCs (osteogenic group) was from the Gene Expression Omnibus database. The analysis was carried out using Bioconductor of R. Gene Set Enrichment Analysis and Kyoto Encyclopedia of Genes and Genomes dataset were applied to identify up- and downregulated signaling pathways. Co-expression network of specific lncRNAs and mRNAs was structured by Cytoscape, while binding sites amongst lncRNA, mRNA, and miRNA were predicted by TargetScan and miRanda. ASCs were derived from human adipose tissue and were authenticated by flow cytometry. ASC cell function was surveyed by alizarin red and alkaline phosphatase (ALP) staining. Molecular mechanism of HIF1A-AS2/miR-665/IL6 axis was investigated by RNAi, cell transfection, western blot, and qRT-PCR. RNA target relationships were validated by dual-luciferase assay. RESULTS: HIF1A-AS2 and IL6 were highly expressed while miR-665 was lowly expressed in induced ASCs. HIF1A-AS2 and IL6 improved the expression level of osteoblast markers Runx2, Osterix, and Osteocalcin and also accelerated the formation of calcium nodule and ALP activity, yet miR-665 had opposite effects. HIF1A-AS2 directly targeted miR-665, whereas miR-665 repressed IL6 expression. Moreover, the HIF1A-AS2/miR-665/IL6 regulating axis activated the PI3K/Akt signaling pathway. CONCLUSIONS: LncRNA HIF1A-AS2 could sponge miR-665 and hence upregulate IL6, activate the PI3K/Akt signaling pathway, and ultimately promote ASC osteogenic differentiation.


Asunto(s)
Tejido Adiposo/metabolismo , Subunidad alfa del Factor 1 Inducible por Hipoxia/genética , Interleucina-6/metabolismo , MicroARNs/genética , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , ARN Largo no Codificante/genética , Células Madre/metabolismo , Diferenciación Celular , Humanos , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , MicroARNs/metabolismo , Osteogénesis , Transducción de Señal , Células Madre/citología , Transfección
12.
Sci Rep ; 7: 41404, 2017 01 23.
Artículo en Inglés | MEDLINE | ID: mdl-28112250

RESUMEN

3ß-Hydroxysteroid-Δ24 reductase (DHCR24), the final enzyme of the cholesterol biosynthetic pathway, has been associated with urogenital neoplasms. However, the function of DHCR24 in endometrial cancer (EC) remains largely elusive. Here, we analyzed the expression profile of DHCR24 and the progesterone receptor (PGR) in our tissue microarray of EC (n = 258), the existing EC database in GEO (Gene Expression Omnibus), and TCGA (The Cancer Genome Atlas). We found that DHCR24 was significantly elevated in patients with EC, and that the up-regulation of DHCR24 was associated with advanced clinical stage, histological grading, vascular invasion, lymphatic metastasis, and reduced overall survival. In addition, DHCR24 expression could be induced by insulin though STAT3, which directly binds to the promoter elements of DHCR24, as demonstrated by ChIP-PCR and luciferase assays. Furthermore, genetically silencing DHCR24 inhibited the metastatic ability of endometrial cancer cells and up-regulated PGR expression, which made cells more sensitive to progestin. Taken together, we have demonstrated for the first time the crucial role of the insulin/STAT3/DHCR24/PGR axis in the progression of EC by modulating the metastasis and progesterone response, which could serve as potential therapeutic targets for the treatment of EC with progesterone receptor loss.


Asunto(s)
Neoplasias Endometriales/enzimología , Neoplasias Endometriales/patología , Endometrio/anomalías , Insulina/efectos adversos , Proteínas del Tejido Nervioso/biosíntesis , Oxidorreductasas actuantes sobre Donantes de Grupo CH-CH/biosíntesis , Regulación hacia Arriba/genética , Enfermedades Uterinas/enzimología , Anciano , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Neoplasias Endometriales/tratamiento farmacológico , Neoplasias Endometriales/genética , Endometrio/enzimología , Endometrio/patología , Inducción Enzimática/efectos de los fármacos , Femenino , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Silenciador del Gen/efectos de los fármacos , Humanos , Acetato de Medroxiprogesterona/farmacología , Acetato de Medroxiprogesterona/uso terapéutico , Persona de Mediana Edad , Invasividad Neoplásica , Proteínas del Tejido Nervioso/genética , Proteínas del Tejido Nervioso/metabolismo , Oxidorreductasas actuantes sobre Donantes de Grupo CH-CH/genética , Oxidorreductasas actuantes sobre Donantes de Grupo CH-CH/metabolismo , Pronóstico , Receptores de Progesterona/genética , Receptores de Progesterona/metabolismo , Factor de Transcripción STAT3/metabolismo , Regulación hacia Arriba/efectos de los fármacos , Enfermedades Uterinas/genética , Enfermedades Uterinas/patología
13.
Exp Cell Res ; 345(1): 93-9, 2016 07 01.
Artículo en Inglés | MEDLINE | ID: mdl-27207585

RESUMEN

Hypertrophic scar (HS) is a fibroproliferative disorder caused by abnormal wound healing, which is characterized by excessive deposition of extracellular matrix (ECM) secreted by fibroblasts. We previous have found that expression of microRNA-21(miR-21) was increased in tissues and fibroblasts of HS. However, the underlying molecular mechanism remains to be further elucidated. In this study, we identified the miR-21 was a marker for the phenotype of HS fibroblasts, as anti-miR-21 reduced expression of fibrosis markers such as Col1A1, Col3A1, Fn and α-SMA in fibroblasts and overexpression of miR-21 promoted fibroproliferative expression in fibroblasts. Furthermore, we also found that miR-21 promoted TGF-ß1 induced fibroproliferative expression by repressing Smad7 expression in vitro. In addition, the miR-21 inhibitor inhibited the growth of hypertrophic scar tissue in vivo (nude mice experimental model). These results indicated that miR-21 was a critical regulator for HS formation and TGF- ß1/miR-21/Smad7 pathway could be a useful therapeutic target for the treatment of HS.


Asunto(s)
Cicatriz Hipertrófica/genética , Cicatriz Hipertrófica/patología , Fibroblastos/metabolismo , Fibroblastos/patología , MicroARNs/metabolismo , Animales , Proliferación Celular , Modelos Animales de Enfermedad , Humanos , Ratones Endogámicos BALB C , Ratones Desnudos , MicroARNs/genética , Proteína smad7/metabolismo , Factor de Crecimiento Transformador beta1/metabolismo , Regulación hacia Arriba/genética
14.
Exp Biol Med (Maywood) ; 241(1): 31-9, 2016 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-26253192

RESUMEN

Tissue-engineered biologic products may be a viable option in the reconstruction of pelvic organ prolapse (POP). This study was based on the hypothesis that human adipose-derived stem cells (hASCs) are viable in acellular bovine pericardium (ABP), when reseeded by two different techniques, and thus, aid in the reconstruction. To investigate the reseeding of hASCs on ABP grafts by using non-invasive bioluminescence imaging (BLI), and to identify the effective hASCs-scaffold combinations that enabled regeneration. Thirty female athymic nude mice were randomly divided into three groups: In the VIVO group, ABPs were implanted in the subcutaneous pockets and enhanced green fluorescent protein luciferase (eGFP·Luc)-hASCs (1 × 10(6) cells/50 µL) were injected on the ABP at the same time. In the VITRO group, the mice were implanted with grafts that ABP were co-cultured with eGFP·Luc-hASCs in vitro. The BLANK group mice were implanted with ABP only. The eGFP·Luc-hASCs reseeded on ABP were analyzed by BLI, histology, and immunohistochemistry. The eGFP·Luc-hASCs reseeded on ABP could be visualized at 12 weeks in vivo. Histology revealed that the VIVO group displayed the highest cell ingrowths, small vessels, and percent of collagen content per unit area. Desmin and α-smooth muscle actin were positive at the same site in the VIVO group cells. However, few smooth muscles were observed in the VITRO and BLANK groups. These results suggest that hASCs reseeded on ABP in vivo during surgery may further enhance the properties of ABP and may promote regeneration at the recipient site, resulting in a promising treatment option for POP.


Asunto(s)
Tejido Adiposo/citología , Pericardio/citología , Células Madre/fisiología , Ingeniería de Tejidos/métodos , Trasplante de Tejidos/métodos , Adulto , Animales , Bovinos , Diferenciación Celular/fisiología , Femenino , Histocitoquímica , Humanos , Mediciones Luminiscentes , Ratones Desnudos , Imagen Óptica , Resultado del Tratamiento
15.
Exp Ther Med ; 9(2): 372-376, 2015 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-25574200

RESUMEN

The aim of the present study was to track in vivo the distribution and survival of adipose-derived mesenchymal stem cells (ASCs) transplanted into female BALB/c nude mice following simulated childbirth injury, using green fluorescent protein and luciferase dual labeling, bioluminescent imaging (BLI) and histological evaluation. The results demonstrated that the dually labeled ASCs could be detected for up to eight weeks in vivo. The number of implanted cells decreased during the first three weeks, and then stabilized until the end of the experiment. According to the linear regression plot, ~27,621 implanted cells survived until eight weeks after implantation. Transplanted ASCs predominantly existed at the inoculation site of the vagina, with little or no spread to other organs. Histological analysis confirmed the survival of the engrafted ASCs. The study provided basic evidence that BLI techniques can be used to monitor ASCs in vivo in real time and in the long term. Through local administration, ASCs could survive in the long term to facilitate repair following pelvic-floor injury.

16.
Biomed Mater ; 10(1): 015005, 2014 Dec 29.
Artículo en Inglés | MEDLINE | ID: mdl-25546879

RESUMEN

Potential scaffolds for repair of the female pelvic floor require new materials and fabrication by novel methods to improve cellular infiltration. An 'ideal' engineered scaffold for pelvic-floor tissue should mimic the three-dimensional (3D) network of the extracellular matrix (ECM), which possesses intricate macro- and nano-architecture. In this study, a series of blended poly(l-lactide-co-ecaprolactone) P(LLA-CL)/thermoplastic polyurethane (TPU) microyarn/microfibrous scaffolds were produced with different weight ratios via dynamic liquid electrospinning and electrospinning. Both biopolymers were dissolved in 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP). Our data showed the mean diameter of microyarn scaffolds to be significantly larger than that of microfibers. Microyarn scaffolds possessed large pore sizes and high porosity. There was no significant difference between the mechanical properties of microyarn and microfibrous scaffolds. Fourier-transform infrared spectroscopy suggested that intermolecular bonds were not present between the molecules of TPU and P(LLA-CL). Morphologic observations using scanning electron microscopy and inverted fluorescence microscopy showed that adipose-derived stem cells labeled with enhanced green fluorescent protein could grow well along or within blend microyarns and migrate within the novel 3D scaffolds. Hematoxylin and eosin staining demonstrated that cell infiltration on microyarn scaffolds was significantly enhanced. The CCK-8 assay showed that microyarns could significantly facilitate cell proliferation compared with microfibrous scaffolds. These results suggested that blend microyarns of P(LLA-CL)/TPU designed to mimic the ECM for female pelvic-floor tissue may be excellent macroporous scaffolds for tissue repair.


Asunto(s)
Materiales Biocompatibles/química , Trastornos del Suelo Pélvico/terapia , Poliuretanos/química , Andamios del Tejido/química , Adipocitos/citología , Adhesión Celular , Proliferación Celular , Matriz Extracelular/metabolismo , Femenino , Proteínas Fluorescentes Verdes/química , Hematoxilina/química , Humanos , Ensayo de Materiales , Microscopía Electrónica de Rastreo , Microscopía Fluorescente , Diafragma Pélvico/fisiopatología , Polímeros/química , Porosidad , Propanoles/química , Espectroscopía Infrarroja por Transformada de Fourier , Células Madre/química , Estrés Mecánico , Resistencia a la Tracción , Ingeniería de Tejidos/métodos , Transfección
17.
Zhonghua Fu Chan Ke Za Zhi ; 45(5): 326-30, 2010 May.
Artículo en Chino | MEDLINE | ID: mdl-20646439

RESUMEN

OBJECTIVE: To identify the morphological characteristics of pelvic diaphragm hiatus in pregnant women with stress urinary incontinence (SUI) by transperineal three-dimensional (3-D) ultrasound. METHODS: From Oct. 2008 to Mar. 2009, 145 pregnant women (third trimester group) at 37-41 weeks of gestation underwent transperineal 3-D ultrasound investigation at Department of Obstetrics and Gynecology, the Sixth People's Hospital, Shanghai Jiaotong University, including 38 pregnant women with stress urinary incontinence (SUI) and the other 107 non SUI pregnant women. In the mean time, 50 normal nulliparous healthy women were chosen as control group. The morphological characteristics of pelvic diaphragm hiatus, the diameter of pelvic diaphragm hiatus, pubovisceral muscle thickness and genitohiatal and levator ani angle were measured at rest, on maximum Valsalva and maximum pelvic floor contraction by 3-D ultrasound, respectively. RESULTS: Loosen connective tissue and pubococcygeus avulsion were observed in some pregnant women at third trimester. The area of pelvic diaphragm hiatus were (15.2+/-1.9), (16.4+/-2.0) and (13.6+/-1.9) cm2, pubovisceral muscle thickness were (0.72+/-0.11), (0.68+/-0.14) and (0.77+/-0.11) cm, levator ani angle were (60+/-8) degrees, (57+/-10) degrees and (64+/-14) degrees at rest, on maximum Valsalva and maximum pelvic floor contraction respectively. These parameters were significantly increased than those in control group [(11.2+/-2.6), (14.5+/-4.5) and (9.2+/-2.6) cm2; (0.66+/-0.10), (0.67+/-0.14) and (0.71+/-0.14) cm; (50+/-4) degrees, (51+/-5) degrees and (46+/-5) degrees] at three maneuvers, respectively (P<0.05). And those parameters of the anteroposterior hiatal diameter, lateral hiatal diameter, perimeter of pelvic diaphragm hiatus and area of pelvic diaphragm hiatus in SUI pregnant women were increased than those in non SUI pregnant women at three maneuvers, respectively (P<0.05). Pubovisceral muscle thickness in SUI pregnant women was significantly lower than that in non SUI pregnant women at maximum pelvic floor contraction (P<0.05), but there were not significant difference between SUI and non SUI pregnant women at rest and on maximum Valsalva in pubovisceral muscle thickness and genitohiatal and levator ani angle (P>0.05). CONCLUSIONS: Pelvic floor anatomic remodeling is identified in late pregnant women. When compared with non pregnant women, the loosen pelvic floor connective tissue and the bigger diameters of pelvic diaphragm are observed in late pregnant women. It is observed that the increased diameters of pelvic diaphragm and decreased thickness of pubovisceral muscle in later pregnant SUI women than those in non SUI pregnant women.


Asunto(s)
Diafragma Pélvico/diagnóstico por imagen , Complicaciones del Embarazo/diagnóstico por imagen , Ultrasonografía/métodos , Incontinencia Urinaria de Esfuerzo/diagnóstico por imagen , Adulto , Estudios de Casos y Controles , Femenino , Humanos , Imagenología Tridimensional/métodos , Contracción Muscular , Diafragma Pélvico/anatomía & histología , Perineo/anatomía & histología , Perineo/diagnóstico por imagen , Embarazo , Complicaciones del Embarazo/fisiopatología , Tercer Trimestre del Embarazo , Estudios Retrospectivos , Incontinencia Urinaria de Esfuerzo/fisiopatología , Adulto Joven
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