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Post-wound infections have remained a serious threat to society and healthcare worldwide. Attempts are still being made to develop an ideal antibacterial wound dressing with high wound-healing potential and strong antibacterial activity against extensively drug-resistant bacteria (XDR). In this study, a biological-based sponge was made from decellularized human placenta (DPS) and then loaded with different concentrations (0, 16 µg/mL, 32 µg/mL, 64 µg/mL) of an antimicrobial peptide (AMP, CM11) to optimize an ideal antibacterial wound dressing. The decellularization of DPS was confirmed by histological evaluations and DNA content assay. The DPS loaded with different contents of antimicrobial peptides (AMPs) showed uniform morphology under a scanning electron microscope (SEM) and cytobiocompatibility for human adipose tissue-derived mesenchymal stem cells. Antibacterial assays indicated that the DPS/AMPs had antibacterial behavior against both standard strain and XDR Acinetobacter baumannii in a dose-dependent manner, as DPS loaded with 64 µg/mL showed the highest bacterial growth inhibition zone and elimination of bacteria under SEM than DPS alone and DPS loaded with 16 µg/mL and 32 µg/mL AMP concentrations. The subcutaneous implantation of all constructs in the animal model demonstrated no sign of acute immune system reaction and graft rejection, indicating in vivo biocompatibility of the scaffolds. Our findings suggest the DPS loaded with 64 µg/mL as an excellent antibacterial skin substitute, and now promises to proceed with pre-clinical and clinical investigations.
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Peptídeos Antimicrobianos , Pele Artificial , Gravidez , Animais , Feminino , Humanos , Placenta , Antibacterianos/farmacologia , Antibacterianos/química , Bandagens , BactériasRESUMO
Extracellular matrix (ECM)-based bioinks has attracted much attention in recent years for 3D printing of native-like tissue constructs. Due to organ unavailability, human placental ECM can be an alternative source for the construction of 3D print composite scaffolds for the treatment of deep wounds. In this study, we use different concentrations (1.5%, 3% and 5%w/v) of ECM derived from the placenta, sodium-alginate and gelatin to prepare a printable bioink biomimicking natural skin. The printed hydrogels' morphology, physical structure, mechanical behavior, biocompatibility, and angiogenic property are investigated. The optimized ECM (5%w/v) 3D printed scaffold is applied on full-thickness wounds created in a mouse model. Due to their unique native-like structure, the ECM-based scaffolds provide a non-cytotoxic microenvironment for cell adhesion, infiltration, angiogenesis, and proliferation. In contrast, they do not show any sign of immune response to the host. Notably, the biodegradation, swelling rate, mechanical property, cell adhesion and angiogenesis properties increase with the increase of ECM concentrations in the construct. The ECM 3D printed scaffold implanted into deep wounds increases granulation tissue formation, angiogenesis, and re-epithelialization due to the presence of ECM components in the construct, when compared with printed scaffold with no ECM and no treatment wound. Overall, our findings demonstrate that the 5% ECM 3D scaffold supports the best deep wound regeneration in vivo, produces a skin replacement with a cellular structure comparable to native skin.
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Objective: Epigenetic and genetic changes have important roles in stem cell achievements. Accordingly, the aim of this
study is the evaluation of the epigenetic and genetic alterations of different culture systems, considering their efficacy in
propagating human spermatogonial stem cells isolated by magnetic-activated cell sorting (MACS).
Materials and Methods: In this experimental study, obstructive azoospermia (OA) patient-derived spermatogonial cells were divided into two groups. The MACS enriched and non-enriched spermatogonial stem cells (SSCs) were cultured in the control and treated groups; co-culture of SSCs with Sertoli cells of men with OA, co-culture of SSCs with healthy Sertoli cells of fertile men, the culture of SSCs on PLA nanofiber and culture of testicular cell suspension. Gene-specific methylation by MSP, expression of pluripotency (NANOG, C-MYC and OCT-4), and germ cells specific genes (Integrin α6, Integrin ß1, PLZF) evaluated. Cultured SSCs from the optimized group were transplanted into the recipient azoospermic mouse.
Results: The use of MACS for the purification of human stem cells was effective at about 69% with the culture of the testicular suspension, being the best culture system. Upon purification, the germ-specific gene expression was significantly higher in testicular cell suspension and treated groups (P≤0.05). During the culture time, gene-specific methylation patterns of the examined genes did not show any changes. Our data from transplantation indicated the homing of the donor-derived cells and the presence of human functional sperm.
Conclusion: Our in vivo and in vitro results confirmed that culture of testicular cell suspension and selection of
spermatogonial cells could be effective ways for purification and enrichment of the functional human spermatogonial cells. The epigenetic patterns showed that the specific methylation of the evaluated genes at this stage remained constant with no alteration throughout the entire culture systems over time.
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INTRODUCTION: It is well documented that some forced exercises can have bad effects on the genital system. Melatonin is a potent antioxidant that is effective in reducing the physical stress.
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Melatonina , Condicionamento Físico Animal , Animais , Antioxidantes/farmacologia , Masculino , Melatonina/farmacologia , Ratos , Espermatogênese , TestículoRESUMO
Improvement of culture system and increasing the proliferation of spermatogonia stem cells under in vitro condition are the essential treatment options for infertility before autologous transplantation. Therefore, the present study aimed to evaluate the proliferation of human spermatogonia stem cells on the electrospun polycaprolactone/gelatin nanocomposite. Therefore, for this purpose, nanofiber porous scaffolds were prepared using the electrospinning method and their structures were then confirmed by SEM. After performing swelling, biodegradability and cell adhesion tests, human spermatogonia stem cells were cultured on scaffolds. In addition, both cell viability and proliferation were assessed using immunocytochemistry, flow cytometry and real-time PCR techniques in culturing during a 3-week period. SEM images indicated the presence of fibres with suitable diameters and arrangement as well as a sufficient porosity in nanocomposite scaffolds, showing good biocompatibility and biodegradability. The results show a significant increase in the number of spermatogonia stem cells in the cultured group on scaffold compared with the control group (p ≤ 0.05). As well, the results show that the expressions of integrin É6 and ß1 and Plzf genes estimated using real-time PCR in nanofiber scaffolds were significantly higher than those of the control group (p ≤ 0.05). However, the expression of c-Kit gene in the 3D group showed a significant decrease compared with the 2D group. Flow cytometry analysis also showed that the number of Plzf-positive cells was significantly higher in nanofiber porous scaffolds compared with the control group (p ≤ 0.05). Additionally, immunocytochemistry findings confirmed the presence of human spermatogonia stem cell colonies. In general, it seems that the designed nanocomposite scaffold could provide a suitable capacity for self-renewal of human spermatogonia stem cells, which can have a good application potential in research and reconstructive medicine related to the field of male infertility.
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Nanofibras , Proliferação de Células , Gelatina , Humanos , Masculino , Nanofibras/química , Células-Tronco , Engenharia Tecidual/métodos , Alicerces Teciduais/químicaRESUMO
BACKGROUND: Therapy based stem cells have offered a novel therapeutic approach for the improvement of neurodegenerative diseases, specially Parkinson. Hence, developing a well-established culture model with appropriate stem cells is extremely crucial in regenerative engineering to provide efficient targeted cells. Human adult mesenchymal stem cells derived from adipose tissue (hADSCs) have emerged as a promising source of stem cells due to their unique potentials of self-renewal and differentiation into other stem cells. The purpose of this study was to investigate the differentiation capacity of hADSCs into dopaminergic and neuron-like cells in the 3D culture plate (Matrigel). METHODS AND MATERIALS: hADSCs were obtained from adipose tissues of patients and then characterized morphologically with flowcytometry. Isolated cells were harvested to perform differentiation on Matrigel and tissue culture plate (TCP) supplemented with induction factors. The survival rate of cells during neural induction was monitored by MTT. The expression of specific cell markers was analyzed by QRT-PCR and immunocytochemistry on days 2, 8 and 14. The level of released dopamine was measured using HPLC technique. RESULTS: Matrigel had a positive effect on maintaining cell growth compared to those on TCP. Moreover, the number of TH and MAPII positive cells is substantially higher in Matrigel than in TCP. Sox2 and Nestin had a prominent expression in hADSCs within the first days of differentiation. The gene expression of neural markers such as TH, Nurr1, LMX1A and DAT was detected and increased after day 8. Moreover, the dopamine released in the cell harvested on Matrigel was greater than those seeded on TCP. CONCLUSIONS: Overall, hADSCs could generate dopaminergic cells, which suggest its strong capability to serve as a tool for Parkinson disease model in the regenerative medicine.
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Colágeno , Neurônios Dopaminérgicos/metabolismo , Laminina , Células-Tronco Mesenquimais/fisiologia , Cultura Primária de Células/métodos , Proteoglicanas , Tecido Adiposo/citologia , Adulto , Diferenciação Celular , Separação Celular , Células Cultivadas , Dopamina/metabolismo , Combinação de Medicamentos , Humanos , Pessoa de Meia-IdadeRESUMO
There are several therapeutic options for spinal cord injury (SCI), among these strategies stem cell therapy is a potential treatment. The stem cells based therapies have been investigating in acute phase of clinical trials for promoting spinal repair in humans through replacement of functional neuronal and glial cells. The aim of this study was to evaluate the differentiation of Human Dental Pulp Stem Cells (hDPSCs) into functional motor neuron like cells (MNLCs) and promote neuroregeneration by stimulating local neurogenesis in the adult spinal cord slice culture. The immunocytochemistry analysis demonstrated that hDPSCs were positive for mesenchymal stem cell markers (CD73, CD90 and CD105) and negative for the hematopoietic markers (CD34 and CD45). hDPSCs were induced to neurospheres (via implementing B27, EGF, and bFGF) and then neural stem cells (NSC). The NSC differentiated into MNLCs in two steps: first by Shh and RA and ; then with GDNF and BDNF administration. The NS and the NSC were assessed for Oct4, nestin, Nanog, Sox2 expression while the MNLCs were evaluated by ISLET1, Olig2, and HB9 genes. Our results showed that hDPSC can be differentiated into motor neuron phenotype with expression of the motor neuron genes. The functionality of MNLCs was demonstrated by FM1-43, intracellular calcium ion shift and co- culture with C2C12. We co-cultivated hDPSCs with adult rat spinal slices in vitro. Immunostaining and hoechst assay showed that hDPSCs were able to migrate, proliferate and integrate in both the anterolateral zone and the edges of the spinal slices.
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Diferenciação Celular , Polpa Dentária/citologia , Células-Tronco/citologia , Células Cultivadas , Humanos , Neurônios Motores/citologia , Células-Tronco Neurais/citologia , Células-Tronco Pluripotentes/citologia , Células-Tronco Pluripotentes/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reprodutibilidade dos Testes , Esferoides Celulares/citologia , Medula Espinal/citologiaRESUMO
OBJECTIVES: Osteoporosis is a major public health problem that is appeared with increasing age. This study evaluated the effect of the algae Dichotomaria obtusata methanol extract on osteogenic differentiation of the cultured bone marrow mesenchymal stem cells (BMMSCs) in vitro and analyzed the algae methanol extract to find out the potent beneficial components. METHODS: Dichotomaria obtusata were collected from the coastal area of Bushehr City in the Persian Gulf, Iran. The expression of osteogenesis-related genes was examined using real-time PCR. The formation of calcium deposits in differentiated MSCs was examined by Alizarin R staining. Analyses of algae extract ingredients were performed by gas chromatography-mass spectrometry (GC-MS). KEY FINDINGS: Methanol extract of the algae caused the up-regulation of osteogenic genes that were significant for Osteopontin and Osteocalcin (P < 0.05) and also led to an increase in calcium deposits and matrix mineralization in BMMSCs. The GC-MS analyses of the algae extracts resulted in the identification of steroids and essential fatty acids. CONCLUSION: The results of the study indicated that the methanol extract of D. obtusata may possess significant potentials for the prevention of osteoporosis in vitro.
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Células-Tronco Mesenquimais/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Extratos Vegetais/farmacologia , Rodófitas/química , Animais , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Cromatografia Gasosa-Espectrometria de Massas , Oceano Índico , Células-Tronco Mesenquimais/citologia , Metanol/química , Camundongos , Camundongos Endogâmicos BALB C , Osteogênese/genética , Osteoporose/prevenção & controle , Regulação para Cima/efeitos dos fármacosRESUMO
Many studies have been devoted to investigating the potential of guided bone regeneration (GBR) membranes for bone defect reconstruction. Regardless of approaches for treating damaged bone tissues, a beneficial therapeutic strategy has remained a challenge. In this study, a novel GBR membrane with polycaprolactone (PCL) and poly(vinyl alcohol) (PVA) containing different concentrations of metformin (Met) for improving osteogenic properties was developed. The membranes were evaluated for their hydrophilicity, degradation rate, swelling ratio, drug release, mechanical properties, and biological responses. The results showed a significant increase in hydrophilicity, swelling ratio, and degradation rate and no significant changes in mechanical properties of PCL/PVA membranes with Met concentration enhancement. A decrease in cell viability cultured on the surface of the PCL/PVA membrane was seen when the amount of Met was changed from 10 to 15 wt %. The results of the in vitro quantitative real-time polymerase chain reaction (qRT-PCR) also confirmed the higher secretion of osteogenic-related genes in a PCL/PVA/Cell/10 wt % Met scaffold than in the PCL/PVA/Cell sample. Therefore, further in vivo studies were conducted using the electrospun PCL/PVA membrane containing human endometrial stem cells (hEnSCs) and 10% Met. Histopathological and histomorphometric results confirmed that PCL/PVA/hEnSCs/10 wt % Met has excellent potential to differentiate hEnSCs into osteogenic lineages and bone regeneration in calvarial defects of rats. The results of this study confirm the high potential of the PCL/PVA/10 wt % Met fibrous membrane preseeded with hEnSCs in GBR applications.
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Metformina , Álcool de Polivinil , Animais , Regeneração Óssea , Feminino , Humanos , Metformina/farmacologia , Osteogênese , Ratos , Células-TroncoRESUMO
PURPOSE: Generating functional gametes for patients with male infertility is of great interest. We investigated different cultural systems for proliferation of SSCs derived from obstructive azoospermic patients. MATERIALS AND METHODS: Testicular cells were obtained from men with obstructive azoospermia. After enzymatic digestion process, cells were assigned to various groups: culture of SSCs in the dish without cover (control group), co-culture of SSCs with infertile Sertoli cells (I), co-culture of SSCs with fertile Sertoli cells (II), culture of SSCs on nanofiber (covered with laminin) (III), culture of testicular cell suspension (IV). Then cells were cultured and colony formation, gene-specific methylation (by MSP), quantitative genes expression of pluripotency (Nanog, C-Myc, Oct-4) and specific germ cell (Integrin α6, Integrin ß1, PLZF) genes were evaluated in five different culture systems. RESULTS: Our findings indicate a significant increase in the number and diameter of colonies in IV group in compare to control group and other groups. Expression of germ specific genes in IV group were significantly increased (P ≤ 0.05) and levels of expression of pluripotency genes were significantly decreased in this group (P ≤ 0.05) compared with other groups. Gene-specific pattern of methylation of examined genes showed no changes in culture systems during the culture era. CONCLUSION: A microenvironment capable of controlling the proliferation of cell colonies can be restored by testicular cell suspension.
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Células-Tronco Germinativas Adultas , Azoospermia , Técnicas de Cultura de Células/métodos , Testículo/citologia , Células Cultivadas , Epigênese Genética , Humanos , MasculinoRESUMO
Human dental pulp stem cells (hDPSCs) could be differentiated into neuron like-cells under particular microenvironments. It has been reported that a wide range of factors, presented in cerebrospinal fluid (CSF), playing part in neuronal differentiation during embryonic stages, we herein introduce a novel culture media complex to differentiate hDPSCs into neuron-like cells. The hDPSCs were initially isolated and characterized. The CSF was prepared from the Cisterna magna of 19-day-old Wistar rat embryos, embryonic cerebrospinal fluid (E-CSF). The hDPSCs were treated by 5% E-CSF for 2 days, then neurospheres were cultured in DMEM/F12 supplemented with 10-6 µm retinoic acid (RA), glial-derived neurotrophic factor and brain-derived neurotrophic factor for 6 days. The cells which were cultured in basic culture medium were considered as control group. Morphology of differentiated cells as well as process elongation were examined by an inverted microscope. In addition, the neural differentiation markers (Nestin and MAP2) were studied employing immunocytochemistry. Neuronal-like processes appeared 8 days after treatment. Neural progenitor marker (Nestin) and a mature neural marker (MAP2) were expressed in treated group. Moreover Nissl bodies were found in the cytoplasm of treated group. Taking these together, we have designed a simple protocol for generating neuron-like cells using CSF from the hDPSCs, applicable for cell therapy in several neurodegenerative disorders including Alzheimer's disease.
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In this study, we synthesized thermo-responsive chitosan (TCTS) hydrogels, and loaded with different concentrations of antimicrobial peptide (AMP) (0, 4, 8 and 16⯵g·ml-1) to fabricate an antibacterial wound dressing against resistant clinical isolates. Physico-chemical properties, release behavior, cytobiocompatibility and antibacterial activity of the AMP-TCTS hydrogels against standard strain and resistant Acinetobacter baumannii were fully determined in vitro. The TCTS-40% ß-glycerolphosphate hydrogels showed a gelation time of 15â¯min at 37⯰C. 80% weight loss at day 35 with no changes in pH value was observed. AMP-TCTS hydrogels showed a burst release of AMP (around 40%) at day 1, and a controlled release up to day 7. A dramatic water uptake was observed at first 4â¯h, and then continued for 10â¯h in a steady manner. All the AMP-TCTS hydrogels showed excellent cytobiocompatibility for human fibroblasts. The TCTS showed no antibacterial activity against both standard strain and clinical isolates. All the AMP-TCTS hydrogels had strong antibacterial activity against standard strains, but only 16⯵g·ml-1 showed antibacterial behavior against resistant A. baumannii. Our results strongly suggest the 16⯵g·ml-1 AMP-TCTS hydrogel as an excellent antibacterial wound dressing against resistant A. baumannii, and now promises to proceed with pre-clinical investigations.
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Antibacterianos/farmacologia , Bandagens , Quitosana/química , Farmacorresistência Bacteriana , Hidrogéis/química , Proteínas Citotóxicas Formadoras de Poros/farmacologia , Acinetobacter baumannii , Materiais Biocompatíveis , Sobrevivência Celular , Células Cultivadas , Sistemas de Liberação de Medicamentos , Fibroblastos , Humanos , Concentração de Íons de Hidrogênio , Testes de Sensibilidade Microbiana , Temperatura , Água/químicaRESUMO
In the current study, electrospun poly(ε-caprolactone)-gelatin (PCL-Gel) fibrous scaffolds containing magnesium oxide (MgO) particles and preseeded with human endometrial stem cells (hEnSCs) were developed to use as wound care material in skin tissue engineering applications. Electrospun fibers were fabricated using PCL-Gel (1:1 [wt/wt]) with different concentrations of MgO particles (1, 2, and 4 wt%). The fibrous scaffolds were evaluated regarding their microstructure, mechanical properties, surface wettability, and in vitro and in vivo performances. The full-thickness excisional wound model was used to evaluate the in vivo wound healing ability of the fabricated scaffolds. Our findings confirmed that the wounds covered with PCL-Gel fibrous scaffolds containing 2 wt% MgO and preseeded with hEnSCs have nearly 79% wound closure ability while sterile gauze showed 11% of wound size reduction. Our results can be employed for biomaterials aimed at the healing of full-thickness skin wounds.
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Endométrio/metabolismo , Gelatina/química , Óxido de Magnésio/química , Poliésteres/química , Pele/metabolismo , Células-Tronco/metabolismo , Alicerces Teciduais/química , Cicatrização , Ferimentos e Lesões , Animais , Feminino , Xenoenxertos , Humanos , Masculino , Porosidade , Ratos , Ratos Wistar , Transplante de Células-Tronco , Ferimentos e Lesões/metabolismo , Ferimentos e Lesões/terapiaRESUMO
The original article was published with incorrect article title.
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Hybrid electrospun fiber containing bioactive molecules, which offer the ability to deliver the cells into the wound bed, will help to achieve a high therapeutic effect. In this study, an electrospun polycaperlactone (PCL) and gelatin (Gela) scaffold containing curcumin loaded chitosan nanoparticle (NCs/Cur) was used to evaluate in vivo wound healing ability of the fabricated scaffolds. The electrospun hybrid scaffold seeded with human endometrial stem cells (EnSCs) showed desirable biocompatibility with the host immune system and wound healing ability in a full-thickness excisional animal model. The constructs were characterized for structural, mechanical and biochemical properties. Fourier transform infrared spectroscopy (FTIR) confirmed all typical absorption characteristics of PCL and Gela polymers as well as NCs and Cur. The results showed the perfect contact angle, wettability and degradability of hybrid fiber scaffolds with the good mechanical and structural characteristics including shape uniformity, pore size and porosity. The cell attachment and proliferation on the PCL/Gela/NCs/Cur was higher than PCL and PCL/Gela scaffolds. In term of the capability of hybrid scaffold and EnSCs in histological analysis, this novel tissue-engineered construct could be suggested as a skin substitute to repair injured skin and regenerative medicine application.
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Quitosana/química , Curcumina/química , Gelatina/química , Nanopartículas/química , Poliésteres/química , Pele/efeitos dos fármacos , Cicatrização/efeitos dos fármacos , Animais , Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Cápsulas , Portadores de Fármacos/química , Liberação Controlada de Fármacos , Eletricidade , Masculino , Teste de Materiais , Fenômenos Mecânicos , Ratos , Ratos Wistar , Pele/fisiopatologia , Alicerces Teciduais , MolhabilidadeRESUMO
PURPOSE: To determine the role of remote perconditioning (RPeC) on renal function and histology in an animal model of unilateral renal ischemia and reperfusion (IR) injury. METHODS: Rats were subjected to 60 min unilateral renal ischemia. RPeC protocol was the application of four cycles of 5 min IR of left femoral artery during renal ischemia. Assessments of histological changes and renal function were made 24 h, 1 week, or 3 weeks later. 99mTc-DMSA scan was performed using a small-animals SPECT system. RESULTS: 24-h reperfusion decreased the 99mTc-DMSA uptake in the left kidney compared to the intact kidney of control animals. RPeC group has higher uptake compared to the IR group. After 1 week and 3 weeks, uptakes were gradually increased in both groups and no differences were observed. Severe morphological changes in the ischemic kidneys of both groups were observed after 24 h which attenuated after 1 week and 3 weeks. Moreover, no differences in creatinine and BUN levels between IR-treated and intact animals were observed. CONCLUSION: These data suggest that RPeC exerts a partially transient improvement in the renal function in the first day after reperfusion. However, long-term follow-up study showed no beneficial effects of RPeC. Moreover, noninvasive 99mTc-DMSA scan revealed a suitable tool in the follow-up evaluation of recovery process in the unilateral renal IR injury models.
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Precondicionamento Isquêmico , Rim/irrigação sanguínea , Traumatismo por Reperfusão/prevenção & controle , Animais , Modelos Animais de Doenças , Precondicionamento Isquêmico/métodos , Estudos Longitudinais , Masculino , Cintilografia , Ratos , Ratos Wistar , Traumatismo por Reperfusão/diagnóstico por imagemRESUMO
Mesenchymal stem cell (MSC) transplantation therapy has been proposed as a promising approach for the treatment of neurodegenerative disease. Chemical and pharmacological preconditioning before transplantation could optimize the therapeutic properties of transplanted MSCs. In this study, we hypothesized that preconditioning treatment with a prolyl hydroxylase inhibitor, dimethyloxalylglycine (DMOG), will increase MSC efficacy and paracrine effects in an amyloid-ß (Aß)-injected Alzheimer rat model. MSCs were incubated in different concentrations of DMOG for 24â¯h. Cell viability, migration, and antioxidant capacity was assessed in DMOG-treated and non-treated MSCs before transplantation into Aß-injected rats. In vitro analysis revealed that DMOG treatment increased cell viability, migration, and expression of CXCR4, CCR2, Nrf2, and HIF-1α in the MSCs. Our in vivo results show that DMOG preconditioning enhances a MSC-mediated rescue of learning and memory function in Aß-injected rats. Furthermore, we found an increased level of BDNF and total antioxidant capacity in the hippocampus of Aß-injected rats following transplantation of preconditioned relative to untreated MSCs. Our results suggest that preconditioning MSCs with DMOG before transplantation may enhance the efficacy of stem cell based therapy in neurodegenerative disease.
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Doença de Alzheimer/terapia , Aminoácidos Dicarboxílicos/uso terapêutico , Sobrevivência Celular/efeitos dos fármacos , Transplante de Células-Tronco Mesenquimais/métodos , Células-Tronco Mesenquimais/efeitos dos fármacos , Doença de Alzheimer/induzido quimicamente , Aminoácidos Dicarboxílicos/farmacologia , Peptídeos beta-Amiloides , Animais , Modelos Animais de DoençasRESUMO
INTRODUCTION: Cell therapy has been widely considered as a therapeutic approach for neurodegenerative diseases and nervous system damage. Cholinergic neurons as one of the most important neurons that play a significant role in controlling emotions, mobility, and autonomic systems. In this study, Human Dental Pulp Stem Cells (hDPSCs) were differentiated into the cholinergic neurons by ß-mercaptoethanol in the preinduction phase and also by the nerve growth factor (NGF) in the induction phase. METHODS: The hDPSCs were evaluated for CD73, CD31, CD34, and Oct-4. Concentration-time relationships for NGF were assessed by evaluating the viability rate of cells and the immune response to nestin, neurofilament 160, microtubule-associated protein-2, and choline acetyltransferase. RESULTS: The hDPSCs had a negative response to CD34 and CD31. The optimal dose for the NGF was 50 ng/mL seven days after the induction when the highest percentage of expressing markers for the Cholinergic neurons (ChAT) was detected. CONCLUSION: The results of this study provided a method for producing cholinergic neurons by hDPSCs, which can be used in cytotherapy for degenerative diseases of the nervous system and also spinal cord injury.