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1.
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
2.
Methods Mol Biol ; 2259: 3-12, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33687705

RESUMO

In the present protocol, extracellular vesicles (EVs) released from a primary culture of human umbilical cord mesenchymal stem cells (MSCs) were isolated by ultracentrifugation processes, characterized by transmission electron microscopy (TEM) and measured by nanoparticle tracking analysis (NTA). Protein was extracted from EVs using RIPA buffer and then was assessed for integrity. The proteomic content of the total EV protein samples was analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) after labeling by tandem mass tag (TMT). This combined approach allowed the development of an effective strategy to study the protein cargo from MSC-derived EVs.


Assuntos
Vesículas Extracelulares/química , Vesículas Extracelulares/ultraestrutura , Células-Tronco Mesenquimais/citologia , Proteínas/análise , Células Cultivadas , Cromatografia Líquida/métodos , Meios de Cultura/química , Humanos , Células-Tronco Mesenquimais/química , Microscopia Eletrônica de Transmissão/métodos , Cultura Primária de Células/métodos , Proteínas/isolamento & purificação , Proteômica/métodos , Espectrometria de Massas em Tandem/métodos , Cordão Umbilical/citologia
3.
Medicine (Baltimore) ; 100(10): e25068, 2021 Mar 12.
Artigo em Inglês | MEDLINE | ID: mdl-33725897

RESUMO

RATIONALE: To describe the clinical effects of human umbilical cord mesenchymal stem cells (UCMSCs) combined with allogenic platelet-rich fibrin (PRF) for the treatment of lower limb ischemia in an elderly patient. PATIENT CONCERNS: The patient was a 93-year-old Chinese woman with bilateral foot gangrene and ulcers lasting for 6 months. She had a prior history of Behcet's disease. DIAGNOSES: The admitting diagnosis for this episode was atherosclerosis bilateral limb ischemia. INTERVENTIONS: First, treatment consisting of immunosuppressants, anticoagulation, antiplatelets, and anti-microbials were instituted. A UCMSC suspension was administered intravenously and injected into the lower limbs twice. An allogenic PRF membrane was externally applied 15 times over the lower limbs. OUTCOMES: The patient's pain improved and the 6 ulcers healed. LESSONS: The combination of UCMSCs with a PRF membrane for the treatment of lower limb ischemia in an elderly patient is effective and safe. More and larger trials are needed before incorporating this therapy into mainstream treatment.


Assuntos
Isquemia/terapia , Transplante de Células-Tronco Mesenquimais/métodos , Doença Arterial Periférica/terapia , Fibrina Rica em Plaquetas , Dedos do Pé/irrigação sanguínea , Administração Intravenosa , Idoso de 80 Anos ou mais , Terapia Combinada/métodos , Feminino , Humanos , Injeções Intralesionais , Isquemia/etiologia , Doença Arterial Periférica/complicações , Transplante Homólogo/métodos , Cordão Umbilical/citologia
4.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 38(2): 117-122, 2021 Feb 10.
Artigo em Chinês | MEDLINE | ID: mdl-33565061

RESUMO

OBJECTIVE: To compare the mRNA level of cell proliferation-related genes Twist1, SIRT1, FGF2 and TGF-ß3 in placenta mesenchymal stem cells (PA-MSCs), umbilical cord mensenchymals (UC-MSCs) and dental pulp mesenchymal stem cells (DP-MSCs). METHODS: The morphology of various passages of PA-MSCs, UC-MSCs and DP-MSCs were observed by microscopy. Proliferation and promoting ability of the three cell lines were detected with the MTT method. Real-time PCR (RT-PCR) was used to determine the mRNA levels of Twist1, SIRT1, FGF2, TGF-ß3. RESULTS: The morphology of UC-MSCs and DP-MSCs was different from that of PA-MSCs. Proliferation ability and promoting ability of the PA-MSCs was superior to that of UC-MSCs and DP-MSCs. In PA-MSCs, expression level of Twist1 and TGF-ß3 was the highest and FGF2 was the lowest. SIRT1 was highly expressed in UC-MSCs. With the cell subcultured, different expression levels of Twist1, SIRT1, FGF2, TGF-ß3 was observed in PA-MSCs, UC-MSCs and DP-MSCs. CONCLUSION: Up-regulated expression of the Twist1, SIRT1 and TGF-ß3 genes can promote proliferation of PA-MSCs, UC-MSCs and DP-MSCs, whilst TGF-ß3 may inhibit these. The regulatory effect of Twist1, SIRT1, FGF2 and TGF-ß3 genes on PA-MSCs, UC-MSCs and DP-MSCs are different.


Assuntos
Proliferação de Células/genética , Fator 2 de Crescimento de Fibroblastos/genética , Células-Tronco Mesenquimais/citologia , Proteínas Nucleares/genética , Sirtuína 1/genética , Fator de Crescimento Transformador beta3/genética , Proteína 1 Relacionada a Twist/genética , Diferenciação Celular , Células Cultivadas , Polpa Dentária/citologia , Feminino , Humanos , Placenta/citologia , Gravidez , Cordão Umbilical/citologia
5.
Int J Mol Sci ; 22(2)2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-33467726

RESUMO

Mesenchymal stem cells (MSCs) have the potential to be a viable therapy against various diseases due to their paracrine effects, such as secretion of immunomodulatory, trophic and protective factors. These cells are known to be distributed within various organs and tissues. Although they possess the same characteristics, MSCs from different sources are believed to have different secretion potentials and patterns, which may influence their therapeutic effects in disease environments. We characterized the protein secretome of adipose (AD), bone marrow (BM), placenta (PL), and Wharton's jelly (WJ)-derived human MSCs by using conditioned media and analyzing the secretome by mass spectrometry and follow-up bioinformatics. Each MSC secretome profile had distinct characteristics depending on the source. However, the functional analyses of the secretome from different sources showed that they share similar characteristics, such as cell migration and negative regulation of programmed cell death, even though differences in the composition of the secretome exist. This study shows that the secretome of fetal-derived MSCs, such as PL and WJ, had a more diverse composition than that of AD and BM-derived MSCs, and it was assumed that their therapeutic potential was greater because of these properties.


Assuntos
Tecido Adiposo/citologia , Células-Tronco Mesenquimais/metabolismo , Placenta/citologia , Cordão Umbilical/citologia , Geleia de Wharton/citologia , Medula Óssea , Células da Medula Óssea/citologia , Diferenciação Celular , Proliferação de Células , Cromatografia Líquida , Análise por Conglomerados , Técnicas de Cocultura , Biologia Computacional , Meios de Cultivo Condicionados , Meios de Cultura Livres de Soro , Feminino , Humanos , Espectrometria de Massas , Osteogênese , Gravidez , Proteômica , Espectrometria de Massas em Tandem
6.
Stem Cells Transl Med ; 10(5): 660-673, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33400390

RESUMO

Acute respiratory distress syndrome (ARDS) in COVID-19 is associated with high mortality. Mesenchymal stem cells are known to exert immunomodulatory and anti-inflammatory effects and could yield beneficial effects in COVID-19 ARDS. The objective of this study was to determine safety and explore efficacy of umbilical cord mesenchymal stem cell (UC-MSC) infusions in subjects with COVID-19 ARDS. A double-blind, phase 1/2a, randomized, controlled trial was performed. Randomization and stratification by ARDS severity was used to foster balance among groups. All subjects were analyzed under intention to treat design. Twenty-four subjects were randomized 1:1 to either UC-MSC treatment (n = 12) or the control group (n = 12). Subjects in the UC-MSC treatment group received two intravenous infusions (at day 0 and 3) of 100 ± 20 × 106 UC-MSCs; controls received two infusions of vehicle solution. Both groups received best standard of care. Primary endpoint was safety (adverse events [AEs]) within 6 hours; cardiac arrest or death within 24 hours postinfusion). Secondary endpoints included patient survival at 31 days after the first infusion and time to recovery. No difference was observed between groups in infusion-associated AEs. No serious adverse events (SAEs) were observed related to UC-MSC infusions. UC-MSC infusions in COVID-19 ARDS were found to be safe. Inflammatory cytokines were significantly decreased in UC-MSC-treated subjects at day 6. Treatment was associated with significantly improved patient survival (91% vs 42%, P = .015), SAE-free survival (P = .008), and time to recovery (P = .03). UC-MSC infusions are safe and could be beneficial in treating subjects with COVID-19 ARDS.


Assuntos
Anti-Inflamatórios/uso terapêutico , Transplante de Células-Tronco Mesenquimais/métodos , Citocinas/sangue , Método Duplo-Cego , Feminino , Humanos , Masculino , Transplante de Células-Tronco Mesenquimais/efeitos adversos , Células-Tronco Mesenquimais , Pessoa de Meia-Idade , Índice de Gravidade de Doença , Resultado do Tratamento , Cordão Umbilical/citologia
7.
ACS Appl Mater Interfaces ; 13(3): 3512-3520, 2021 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-33435676

RESUMO

Near-infrared conjugated polymer nanoparticles (NIR-CPNs) have been widely used in in vivo imaging fields. However, most of them face the aggregation-induced fluorescence quenching (ACQ) dilemma and serious dye leakage behavior, which impedes the long-term monitoring of transplanted cells in vivo. In the present work, a novel strategy of sandwich-type encapsulation of the conjugated polymer interlayer in the crystalline SiO2 core + shell (SSiO2@SPFTBT@CSiO2) is developed, which works well to avoid the ACQ problem by homogeneously dispersing poly((9,9-dioctylfluorene-2,7-diyl)-alt-(4,7-di(thiophene-2-yl)-2,1,3-benzothiadiazole)-5',5″-diyl) (PFTBT) and suppressing intermolecular π-π stacking. Furthermore, the unparalleled nanostructure efficiently stabilizes nanoparticles and successfully achieves long-term biocompatibility without interfering the biological characteristics of stem cells, indicating the potential of SSiO2@SPFTBT@CSiO2 in cell labeling. In addition, the fate of human umbilical cord mesenchymal stem cells (hucMSCs) in a mouse model with acute liver injury was disclosed. We found that the hucMSCs mainly migrated from the lungs to the injured liver and most transplanted hucMSCs were cleared up by the liver at 8 days post-injection. Revelation of the shuttle process and period will benefit in improving the clinical efficacy of hucMSCs, and the sandwich-type encapsulation strategy could also open a new avenue to obtain bright and robust NIR-CPNs for long-term fluorescence imaging.


Assuntos
Rastreamento de Células/métodos , Células-Tronco Mesenquimais/citologia , Nanopartículas/química , Imagem Óptica/métodos , Polímeros/química , Tiofenos/química , Lesão Pulmonar Aguda/diagnóstico por imagem , Lesão Pulmonar Aguda/terapia , Animais , Linhagem Celular , Humanos , Masculino , Transplante de Células-Tronco Mesenquimais , Camundongos Endogâmicos BALB C , Dióxido de Silício/química , Tiadiazóis/química , Cordão Umbilical/citologia
8.
Stem Cell Res Ther ; 12(1): 91, 2021 01 29.
Artigo em Inglês | MEDLINE | ID: mdl-33514427

RESUMO

BACKGROUND: Acute respiratory distress syndrome (ARDS) is a fatal complication of coronavirus disease 2019 (COVID-19). There are a few reports of allogeneic human mesenchymal stem cells (MSCs) as a potential treatment for ARDS. In this phase 1 clinical trial, we present the safety, feasibility, and tolerability of the multiple infusions of high dose MSCs, which originated from the placenta and umbilical cord, in critically ill COVID-19-induced ARDS patients. METHODS: A total of 11 patients diagnosed with COVID-19-induced ARDS who were admitted to the intensive care units (ICUs) of two hospitals enrolled in this study. The patients were critically ill with severe hypoxemia and required mechanical ventilation. The patients received three intravenous infusions (200 × 106 cells) every other day for a total of 600 × 106 human umbilical cord MSCs (UC-MSCs; 6 cases) or placental MSCs (PL-MSCs; 5 cases). FINDINGS: There were eight men and three women who were 42 to 66 years of age. Of these, six (55%) patients had comorbidities of diabetes, hypertension, chronic lymphocytic leukemia (CLL), and cardiomyopathy (CMP). There were no serious adverse events reported 24-48 h after the cell infusions. We observed reduced dyspnea and increased SpO2 within 48-96 h after the first infusion in seven patients. Of these seven patients, five were discharged from the ICU within 2-7 days (average: 4 days), one patient who had signs of acute renal and hepatic failure was discharged from the ICU on day 18, and the last patient suddenly developed cardiac arrest on day 7 of the cell infusion. Significant reductions in serum levels of tumor necrosis factor-alpha (TNF-α; P < 0.01), IL-8 (P < 0.05), and C-reactive protein (CRP) (P < 0.01) were seen in all six survivors. IL-6 levels decreased in five (P = 0.06) patients and interferon gamma (IFN-γ) levels decreased in four (P = 0.14) patients. Four patients who had signs of multi-organ failure or sepsis died in 5-19 days (average: 10 days) after the first MSC infusion. A low percentage of lymphocytes (< 10%) and leukocytosis were associated with poor outcome (P = 0.02). All six survivors were well with no complaints of dyspnea on day 60 post-infusion. Radiological parameters of the lung computed tomography (CT) scans showed remarkable signs of recovery. INTERPRETATION: We suggest that multiple infusions of high dose allogeneic prenatal MSCs are safe and can rapidly improve respiratory distress and reduce inflammatory biomarkers in some critically ill COVID-19-induced ARDS cases. Patients that develop sepsis or multi-organ failure may not be good candidates for stem cell therapy. Large randomized multicenter clinical trials are needed to discern the exact therapeutic potentials of MSC in COVID-19-induced ARDS.


Assuntos
/terapia , Transplante de Células-Tronco Mesenquimais , /terapia , Adulto , Idoso , Biomarcadores/sangue , Comorbidade , Cuidados Críticos , Estado Terminal , Feminino , Humanos , Hipóxia/virologia , Inflamação , Unidades de Terapia Intensiva , Pulmão/diagnóstico por imagem , Masculino , Células-Tronco Mesenquimais/citologia , Pessoa de Meia-Idade , Segurança do Paciente , Placenta/citologia , Gravidez , Respiração Artificial , Sepse/virologia , Tomografia Computadorizada por Raios X , Transplante Homólogo , Resultado do Tratamento , Cordão Umbilical/citologia
9.
Zhonghua Nan Ke Xue ; 26(6): 564-569, 2020 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-33356048

RESUMO

Mesenchymal stem cells (MSC), as a type of multifunction cells capable of self-renewal and multi-directional differentiation and with a low immunogenicity, have been widely applied in life sciences and medicine in recent years. Male infertility is very complicated pathogenically and there is no ideal method for its treatment. The potential ability of multi-differentiation and paracrine function of MSCs is bringing new hope to patients with male infertility. MSCs can be derived from the bone marrow, adipose tissue, and umbilical cord, on which lots of studies, either in vitro or in vivo, have been conducted and achieved satisfactory results. Many findings indicate that MSCs can differentiate into germ-like cells if specifically induced or transplanted into the testis, improve the local microenvironment of spermatogenesis and reconstruct the spermatogenic process by secreting nutritional factors after transplantation. This article presents an overview of the advances in the studies and application prospects of MSCs in male infertility.


Assuntos
Infertilidade Masculina , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/citologia , Tecido Adiposo/citologia , Células da Medula Óssea , Diferenciação Celular , Humanos , Infertilidade Masculina/terapia , Masculino , Cordão Umbilical/citologia
10.
Aging (Albany NY) ; 13(2): 2231-2250, 2020 12 09.
Artigo em Inglês | MEDLINE | ID: mdl-33318310

RESUMO

Myocardial infarction is a cardiovascular disease with high mortality. Human umbilical cord mesenchymal stem cells (hUC-MSCs) with strong self-renewal capacity and multipotency, provide the possibility of replacing injured cardiomyocytes. hUC-MSCs were cultured on polyacrylamide hydrogels with stiffnesses corresponding to Young's modulus of 13-16kPa and 62-68kPa which mimic the stiffnesses of healthy heart tissue and fibrotic myocardium. The expression of early myocardial markers Nkx2.5, GATA4, Mesp1 and the mature myocardial markers cTnT, cTnI, α-actin were detected by RT-PCR and Western Blot, which showed that soft matrix (13-16 kPa) tended to induce the differentiation of hUC-MSCs into myocardium, compared with stiff matrix (62-68 kPa). Piezos are mechanically sensitive non-selective cation channels. The expression of Piezo1 increased with the stiffness gradient of 1-10kPa, 13-16kPa, 35-38kPa and 62-68kPa on the 1st day, but Piezo2 expression was irregular. The expression of integrin ß1 and calcium ions were also higher on stiff substrate than on soft substrate. hUC-MSCs tend to differentiate into myocardium on the matrix stiffness of 13-16 kPa. The relationship among matrix stiffness, Piezo1 and myocardial differentiation needs further validation.


Assuntos
Diferenciação Celular/fisiologia , Matriz Extracelular/metabolismo , Células-Tronco Mesenquimais/citologia , Miocárdio/citologia , Miócitos Cardíacos/metabolismo , Cordão Umbilical/citologia , Células Cultivadas , Módulo de Elasticidade , Feminino , Humanos , Hidrogéis , Células-Tronco Mesenquimais/metabolismo , Infarto do Miocárdio/metabolismo , Miocárdio/metabolismo , Cordão Umbilical/metabolismo
11.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 36(10): 897-902, 2020 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-33148384

RESUMO

Objective To compare the efficiency of four methods for extracting extracellular vesicles (EVs) from human umbilical cord mesenchymal stem cells(hUCMSCs). Methods EVs were isolated from the conditioned medium of hUCMSCs by ultracentrifugation (group A), or ultrafiltration combined with ultracentrifugation (group B), or ultrafiltration combined with polyethylene glycol precipitation (group C), or ultrafiltration combined with aqueous two phase system (group D). The total protein concentration of EVs in each group was determined by BCA method. The expression of Alix, CD9, and calnexin were detected by Western blotting. The morphology of EVs was analyzed by transmission electron microscopy. The particle size distribution and particle concentration of EVs were measured by nanoparticle tracking analysis. Results The total protein concentrations of EVs extracted by the above four methods were (1.92±1.77) µg/µL, (18.1±1.07) µg/µL, (6.33±1.02) µg/µL, (36.48±23.13) µg/µL from group A to D respectively. We observed the expression of CD9 and Alix, but not calnexin, in EVs from group A, B and C. However, the expression levels of CD9 and Alix were lowest in group C. In addition, the expression of CD9, Alix and calnexin were undetectable in EVs from group D. The particle concentrations of EVs in group A, B and C were 0.85×1011 particles/mL, 0.63×1011 particles/mL, 1.83×1011 particles/mL, respectively. Meanwhile, the particle distributions were all within the size range of EVs. We also observed the typical saucer-like membrane structure in EVs from group A, B and C. Conclusion The method of ultrafiltration combined with ultracentrifugation could be applied to the experiments demanding large amounts of EVs. The method of ultracentrifugation is recommended for the extraction of little amounts of EVs due to the lower risk of EV fragmentation.


Assuntos
Meios de Cultivo Condicionados , Vesículas Extracelulares , Células-Tronco Mesenquimais , Humanos , Microscopia Eletrônica de Transmissão , Ultracentrifugação , Ultrafiltração , Cordão Umbilical/citologia
12.
Cell Transplant ; 29: 963689720965980, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33040594

RESUMO

Novel therapies are urgently needed to combat the severe cytokine storm syndromes induced by coronavirus disease 2019 (COVID-19). An increasing number of preclinical and clinical investigations of stem cell and derivatives therapy for COVID-19 were being carried out, among which several studies have preliminarily demonstrated the safety and possible efficacy of stem cell transplantation therapy, providing a hint to solve the tricky situation of anti-COVID-19.


Assuntos
Infecções por Coronavirus/terapia , Citocinas/metabolismo , Transplante de Células-Tronco Mesenquimais , Pneumonia Viral/terapia , Betacoronavirus/isolamento & purificação , Infecções por Coronavirus/patologia , Infecções por Coronavirus/virologia , Humanos , Linfócitos/citologia , Linfócitos/metabolismo , Pandemias , Peptidil Dipeptidase A/genética , Peptidil Dipeptidase A/metabolismo , Pneumonia Viral/patologia , Pneumonia Viral/virologia , Cordão Umbilical/citologia
13.
Stem Cell Res Ther ; 11(1): 361, 2020 08 18.
Artigo em Inglês | MEDLINE | ID: mdl-32811531

RESUMO

BACKGROUND: COVID-19 is a highly infectious respiratory disease. No therapeutics have yet been proven effective for treating severe COVID-19. OBJECTIVES: To determine whether human umbilical cord mesenchymal stem cell infusion may be effective and safe for the treatment of severe COVID-19. METHODS: Patients with severe COVID-19 were randomly divided into 2 groups: the standard treatment group and the standard treatment plus hUC-MSC infusion group. The incidence of progression from severe to critical illness, 28-day mortality, clinical symptom improvement, time to clinical symptom improvement, hematologic indicators including C-reactive protein, lymphocyte number, and interleukin 6, and imaging changes were observed and compared between the two groups. MEASUREMENTS AND MAIN RESULTS: The incidence of progression from severe to critical illness and the 28-day mortality rate were 0 in the hUC-MSC treatment group, while 4 patients in the control group deteriorated to critical condition and received invasive ventilation; 3 of them died, and the 28-day mortality rate was 10.34%. In the hUC-MSC treatment group, the time to clinical improvement was shorter than that in the control group. Clinical symptoms of weakness and fatigue, shortness of breath, and low oxygen saturation obviously improved beginning on the third day of stem cell infusion and reached a significant difference on day 7. CRP and IL-6 levels were significantly lower from day 3 of infusion, the time for the lymphocyte count to return to the normal range was significantly faster, and lung inflammation absorption was significantly shorter on CT imaging in the hUC-MSC group than in the control group. CONCLUSIONS: Intravenous transplantation of hUC-MSCs is a safe and effective method that can be considered a salvage and priority treatment option for severe COVID-19. TRIAL REGISTRATION: Chinese Clinical Trial Registration; ChiCTR2000031494; Registered on 2 April 2020; http:// www.medresman.org.


Assuntos
Infecções por Coronavirus/terapia , Transplante de Células-Tronco Mesenquimais , Pneumonia Viral/terapia , Cordão Umbilical/citologia , Adulto , Idoso , Betacoronavirus/isolamento & purificação , Betacoronavirus/patogenicidade , Proteína C-Reativa/metabolismo , China , Infecções por Coronavirus/mortalidade , Infecções por Coronavirus/virologia , Feminino , Humanos , Interleucina-6/metabolismo , Contagem de Linfócitos , Masculino , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Pessoa de Meia-Idade , Pandemias , Pneumonia Viral/mortalidade , Pneumonia Viral/virologia , Taxa de Sobrevida , Tórax/diagnóstico por imagem , Tomografia Computadorizada por Raios X , Transplante Homólogo , Resultado do Tratamento
14.
Int J Nanomedicine ; 15: 5911-5926, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32848396

RESUMO

Purpose: Chronic refractory wounds are a multifactorial comorbidity of diabetes mellitus with the characteristic of impaired vascular networks. Currently, there is a lack of effective treatments for such wounds. Various types of mesenchymal stem cell-derived exosomes (MSC-exos) have been shown to exert multiple therapeutic effects on skin regeneration. We aimed to determine whether a constructed combination of human umbilical cord MSC (hUCMSC)-derived exosomes (hUCMSC-exos) and Pluronic F-127 (PF-127) hydrogel could improve wound healing. Materials and Methods: We topically applied human umbilical cord-derived MSC (hUCMSC)-derived exosomes (hUCMSC-exos) encapsulated in a thermosensitive PF-127 hydrogel to a full-thickness cutaneous wound in a streptozotocin-induced diabetic rat model. The material properties and wound healing ability of the hydrogel and cellular responses were analyzed. Results: Compared with hUCMSC-exos, PF-127-only or control treatment, the combination of PF-127 and hUCMSC-exos resulted in a significantly accelerated wound closure rate, increased expression of CD31 and Ki67, enhanced regeneration of granulation tissue and upregulated expression of vascular endothelial growth factor (VEGF) and factor transforming growth factor beta-1 (TGFß-1). Conclusion: The efficient delivery of hUCMSC-exos in PF-127 gel and improved exosome ability could promote diabetic wound healing. Thus, this biomaterial-based exosome therapy may represent a new therapeutic approach for cutaneous regeneration of chronic wounds.


Assuntos
Diabetes Mellitus Experimental/tratamento farmacológico , Exossomos/química , Hidrogéis/farmacologia , Células-Tronco Mesenquimais/citologia , Poloxâmero/química , Cicatrização/efeitos dos fármacos , Animais , Diabetes Mellitus Experimental/complicações , Pé Diabético/tratamento farmacológico , Pé Diabético/patologia , Células Endoteliais da Veia Umbilical Humana , Humanos , Hidrogéis/química , Masculino , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Ratos Sprague-Dawley , Regeneração , Pele/efeitos dos fármacos , Pele/lesões , Fenômenos Fisiológicos da Pele , Estreptozocina , Cordão Umbilical/citologia , Fator A de Crescimento do Endotélio Vascular/genética , Cicatrização/fisiologia
15.
Medicine (Baltimore) ; 99(28): e21222, 2020 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-32664175

RESUMO

Cervus and cucumis peptides (Lugua polypeptides, LG) are traditional Chinese medicine, which are active components of polypeptide extracted from Sika deer bone and melon seed, and they contain bone induced polypeptide biological factors. Umbilical cord mesenchymal stem cell, (UC-MSC) have tissue repair multiple effects, anti-inflammatory, and immune regulation function, which become a very promising start in rheumatoid arthritis (RA) treatment. Hence, LG combined UC-MSC can significantly enhance the UC-MSC treatment of rheumatoid arthritis (RA).To explore the clinical curative effect and therapeutic mechanism of LG combined UC-MSC for treating RA.119 patients were divided into control and treatment groups, and both groups were treated with methotrexate tablets, leflunomide, and UC-MSC. But, LG were added to the treatment group. In vitro, the effects of LG on UC-MSC cell secretion of anti-inflammatory factors were also performed.The Health Assessment Questionnaire; the 28 joint disease activity score; C reactive protein; the erythrocyte sedimentation rate; rheumatoid factor; and anti-cyclic citrullinated peptide antibody were significantly reduced in treatment group 1 year after treatment (P < .05). In vitro, compared with the control group, the number of hepatocyte growth factor (HGF), the secretion of prostaglandin E2 (PGE2) and tumor necrosis factor-inducible gene 6 protein (TSG6) increased significantly (P < .05).LG combined UC-MSCs can significantly improve the curative effect of RA patients, while LG may reduce inflammatory cytokines, regulate immunity, improve microcirculation, and are conducive to UC-MSCs migration and the repair of damaged tissue.


Assuntos
Artrite Reumatoide/terapia , Transplante de Células-Tronco Mesenquimais/métodos , Células-Tronco Mesenquimais/química , Adulto , Animais , Artrite Reumatoide/sangue , Sedimentação Sanguínea , Proteína C-Reativa/análise , Cucumis , Cervos , Feminino , Fator de Crescimento de Hepatócito/sangue , Humanos , Masculino , Pessoa de Meia-Idade , Fator Reumatoide/sangue , Resultado do Tratamento , Cordão Umbilical/citologia
17.
Zhejiang Da Xue Xue Bao Yi Xue Ban ; 40(7): 988-994, 2020 May 25.
Artigo em Chinês | MEDLINE | ID: mdl-32701245

RESUMO

OBJECTIVE: To study the anti- fibrotic effect of human umbilical cord mesenchymal stem cell-derived exosomes (hUCMSC-EXOs) and explore the mechanism. METHODS: Twenty-four C57 BL/6 mice were divided into 4 groups (n=6), including the control group treated with intratracheal injection of saline (3 mg/kg); lung fibrosis model group with intratracheal injection of 1.5 mg/mL bleomycin solution (prepared with saline, 3 mg/kg); EXOs1 group with intratracheal injection of 1.5 mg/mL bleomycin solution (3 mg/kg) and hUCMSC-EXOs (100 µg/250 µL, given by tail vein injection on the next day after modeling); and EXOs2 group with intratracheal injection of 1.5 mg/mL bleomycin solution (3 mg/kg) and hUCMSC-EXOs (100 µg/250 µL, given by tail vein injection on the 10th day after modeling). At 21 days after modeling, pulmonary index, lung tissue pathology and collagen deposition in the mice were assessed using HE staining and Masson staining. The expression level of TGF-ß1 was detected using ELISA, and vimentin, E-cadherin and phosphorylated Smad2/3 (p-Smad2/3) were detected using immunohistochemical staining. CCK8 assay was used to evaluate the effect of hUCMSCEXOs on the viability of A549 cells, and Western blotting was used to detect the expression levels of p-Smad2/3, vimentin, and E-cadherin in the cells. RESULTS: Compared with those in the model group, the mice treated with hUCMSC-EXOs showed significantly reduced the pulmonary index (P < 0.05), collagen deposition, lung tissue pathologies, lowered expressions of TGF-ß1 (P < 0.05), vimentin, and p-Smad2/3 and increased expression of E-cadherin. hUCMSC-EXOs given on the second day produced more pronounced effect than that given on the 11th day (P < 0.05). CCK8 assay results showed that hUCMSC-EXOs had no toxic effects on A549 cells (P > 0.05). Western blotting results showed that hUCMSC-EXOs treatment significantly increased the expression of E-cadherin and decreased the expressions of p-Smad2/3 and vimentin in the cells. CONCLUSIONS: hUCMSC-EXOs can alleviate pulmonary fibrosis in mice by inhibiting epithelialmesenchymal transition activated by the TGF-ß1/Smad2/3 signaling pathway, and the inhibitory effect is more obvious when it is administered on the second day after modeling.


Assuntos
Exossomos , Células-Tronco Mesenquimais , Fibrose Pulmonar , Animais , Transição Epitelial-Mesenquimal , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Humanos , Células-Tronco Mesenquimais/citologia , Camundongos , Fibrose Pulmonar/terapia , Fator de Crescimento Transformador beta1/genética , Cordão Umbilical/citologia
18.
Cell Prolif ; 53(8): e12862, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32597552

RESUMO

OBJECTIVE: Longitudinal studies have indicated VCAM-1+ mesenchymal stem/stromal cells (MSCs) as promising resources in regenerative medicine, yet the abundance in gene expression is far from adequate in the advantaged and "discarded" hUC-MSCs. Thus, high-efficient preparation and systematic dissection of the signatures and biofunctions of the subpopulation is the prerequisite for large-scale clinical applications. MATERIALS AND METHODS: We primarily took advantage of a cytokine-based programming strategy for large-scale VCAM-1+ hUC-MSC generation (III-MSCs). Thereafter, we conducted multifaceted analyses including cytomorphology, immunophenotype, cell vitality, multilineage differentiation, whole-genome analysis, tube formation and Matrigel plug assay, lymphocyte activation and differentiation, and systemic transplantation for aplastic anaemia (AA) treatment. RESULTS: III-MSCs with high-proportioned VCAM-1 expression were obtained by combining IL-1ß, IL-4 with IFN-γ, which exhibited comparable immunophenotype with untreated hUC-MSCs (NT-MSCs) but revealed multidimensional superiorities both at the cellular and molecular levels. Simultaneously, systemic infusion of III-MSCs could significantly ameliorate clinicopathological features and finally help facilitate haematopoietic reconstruction and immunoregulation in AA mice. CONCLUSIONS: We have established a high-efficient procedure for large-scale generation of III-MSCs with preferable signatures and efficacy upon aplastic anaemia in mice. Our findings suggested that III-MSCs were advantageous sources with multifaceted characteristics for regenerative medicine.


Assuntos
Anemia Aplástica/terapia , Células-Tronco Mesenquimais/citologia , Cordão Umbilical/citologia , Molécula 1 de Adesão de Célula Vascular/metabolismo , Animais , Diferenciação Celular/fisiologia , Proliferação de Células/fisiologia , Células Cultivadas , Imunofenotipagem/métodos , Transplante de Células-Tronco Mesenquimais/métodos , Camundongos
19.
Medicine (Baltimore) ; 99(25): e20628, 2020 Jun 19.
Artigo em Inglês | MEDLINE | ID: mdl-32569193

RESUMO

Traditional Chinese medicines are used in promotion of fractured bone healing and bone diseases. Some studies reported total flavonoids from plant can be used as an auxiliary source of exogenous.Use different methods to identify and verify effects of total flavonoids from Arachniodes exilis (TFAE) on human umbilical cord mesenchymal stem cells (HUCMSCs) in vitro.Concentrations of 1 and 5 µg/mL TFAE significantly increased ALPase activity in HUCMSCs compared to the other concentrations at days 3 and 7 (P < .05). RT-PCR showed that expression levels of osteogenic genes (Col1a1, OPN, Runx2 and Osx) were remarkably enhanced in HUCMSCs following treatment with different concentrations of TFAE for 9 days compared with 0 µg/mL TFAE group (control). The results showed that concentration < 5 µg/mL of TFAE induced osteogenic differentiation in HUCMSCs Alizarin red staining assays revealed that both TFAE and S1191 was significantly decreased (7.80 ±â€Š0.66) compared with the TFAE group (16.00 ±â€Š0.97) (P < .01). ALPase activity on days 3 and 7 was relatively lower in HUCMSCs grown in media supplemented with both S1191 and TFAE than that of in TFAE group only. The results indicated that osteogenic markers (Col1a1, OPN, Runx2 and Osx) were significantly downregulated in the TFAE + S1191 group in comparison to the control group. The expressions of Col1a and OPN in the TFAE + S1191 group decreased significantly (P < .01) by Western blotting.TFAE promotes the odonto/osteogenic differentiation of human UCMSCs via activation of ER.


Assuntos
Flavonoides/farmacologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Extratos Vegetais/farmacologia , Plantas Medicinais/química , Cordão Umbilical/citologia , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Humanos , Técnicas In Vitro , Medicina Tradicional Chinesa
20.
Exp Mol Pathol ; 115: 104468, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32445750

RESUMO

OBJECTIVE: Exosomes originated from mesenchymal stem cells (MSCs) benefit wound healing. This study investigated effects of exosomes originated from human umbilical cord MSCs (hUC-MSCs) on dermal fibroblasts-myofibroblasts transition via the TGF-ß1/Smad2/3 signaling pathway. METHODS: Firstly, hUC-MSCs were collected and identified. Alizarin red, oil red O staining and toluidine blue staining were used to determine the osteogenic, adipogenic and chondrogenic differentiation abilities of hUC-MSCs. Then exosomes from hUC-MSCs were extracted and identified. To figure out the roles of exosomes and TGF-ß1 in dermal fibroblasts-myofibroblasts transition, dermal fibroblasts were treated with TGF-ß1 or/and exosomes at different concentrations. RT-qPCR, Western blot analyses were employed to examine levels of Collagen I, Collagen III, α-smooth muscle actin (α-SMA), and Smad2/3 phosphorylation, and immunofluorescence was employed to test α-SMA content and the localization and nucleation of Smad2/3 protein in cells. RESULTS: hUC-MSCs and exosomes were successfully cultured and extracted. Levels of Collagen I, Collagen III, α-SMA, and Smad2/3, and Smad2/3 phosphorylation in fibroblasts treated with exosomes decreased markedly. After treatment with exosomes and TGF-ß1 together, levels of Collagen I, Collagen III, α-SMA, and Smad2/3, and Smad2/3 phosphorylation in fibroblasts decreased significantly as compared to TGF-ß1-treated fibroblasts. Exosome treatment reduced the entry of Smad2/3 into fibroblasts. CONCLUSION: Our data suggested that hUC-MSCs-derived exosomes could inhibit dermal fibroblasts-myofibroblasts transition by inhibiting the TGF-ß1/Smad2/3 signaling pathway.


Assuntos
Exossomos/metabolismo , Células-Tronco Mesenquimais/metabolismo , Miofibroblastos/metabolismo , Transdução de Sinais , Proteínas Smad/metabolismo , Fator de Crescimento Transformador beta1/metabolismo , Cordão Umbilical/citologia , Células Cultivadas , Derme/citologia , Exossomos/ultraestrutura , Humanos , Recém-Nascido
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