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1.
Arch. endocrinol. metab. (Online) ; 65(3): 342-351, May-June 2021. tab, graf
Article in English | LILACS | ID: biblio-1285166

ABSTRACT

ABSTRACT Objective: Adipose tissue-derived stromal/stem cells (ASCs) and vitamin D have immunomodulatory actions that could be useful for type 1 diabetes (T1D). We aimed in this study to investigate the safety and efficacy of ASCs + daily cholecalciferol (VIT D) for 6 months in patients with recent-onset T1D. Materials and methods: In this prospective, dual-center, open trial, patients with recent onset T1D received one dose of allogenic ASC (1 x 106 cells/kg) and cholecalciferol 2,000 UI/day for 6 months (group 1). They were compared to patients who received chol-ecalciferol (group 2) and standard treatment (group 3). Adverse events were recorded; C-peptide (CP), insulin dose and HbA1c were measured at baseline (T0), after 3 (T3) and 6 months (T6). Results: In group 1 (n = 7), adverse events included transient headache (all), mild local reactions (all), tachycardia (n = 4), abdominal cramps (n = 1), thrombophlebitis (n = 4), scotomas (n = 2), and central retinal vein occlusion at T3 (n = 1, resolution at T6). Group 1 had an increase in basal CP (p = 0.018; mean: 40.41+/-40.79 %), without changes in stimulated CP after mixed meal (p = 0.62), from T0 to T6. Basal CP remained stable in groups 2 and 3 (p = 0.58 and p = 0.116, respectively). Group 1 had small insulin requirements (0.31+/- 0.26 UI/kg) without changes at T6 (p = 0.44) and HbA1c decline (p = 0.01). At T6, all patients (100%; n = 7) in group 1 were in honeymoon vs 75% (n = 3/4) and 50% (n = 3/6) in groups 2 and 3, p = 0.01. Conclusions: Allogenic ASC + VIT D without immunosuppression was safe and might have a role in the preservation of β-cells in patients with recent-onset T1D. ClinicalTrials.gov: NCT03920397.


Subject(s)
Humans , Stem Cells/cytology , Cholecalciferol/therapeutic use , Mesenchymal Stem Cell Transplantation , Diabetes Mellitus, Type 1/drug therapy , Pilot Projects , Adipose Tissue/cytology , Prospective Studies
2.
Braz. j. med. biol. res ; 54(2): e9944, 2021. tab, graf
Article in English | LILACS, ColecionaSUS | ID: biblio-1142581

ABSTRACT

The aim of this study was to inhibit adipogenic differentiation by transfecting two growth factors, platelet-derived growth factor (PDGF-BB) and bone morphogenic protein 2 (BMP-2), into modified rat bone marrow mesenchymal stem cells (rBMSCs) and then compounded with platelet-rich plasma (PRP). To achieve rBMSCs, the osteoporosis model of rats was established, and then the rBMSCs from the rats were isolated and identified. Co-transfection of rBMSCs with PDGF-BB-GFP and BMP-2 and detection of PDGF-BB/BMP-2 expression in transfected BMSCs was assessed by qRT-PCR and western blot, respectively. Moreover, the effect of the two growth factors transfection of rBMSCs on adipogenic differentiation was evaluated by oil red O staining and western blot, respectively. Finally, construction of the two growth factors transfection of rBMSCs compounded with PRP and detection of adipogenic differentiation were assessed by oil red O staining, CCK-8, and western blot, respectively. In vitro studies revealed that the two growth factors transfection of rBMSCs compounded with PRP promoted cell viability and inhibited adipogenic differentiation and could be promising for inhibiting adipogenic differentiation.


Subject(s)
Animals , Rats , Cell Differentiation , Adipose Tissue/cytology , Platelet-Rich Plasma , Bone Morphogenetic Protein 2/genetics , Mesenchymal Stem Cells/cytology , Becaplermin/genetics , Transfection , Cells, Cultured
3.
Acta Academiae Medicinae Sinicae ; (6): 755-765, 2020.
Article in Chinese | WPRIM | ID: wpr-878674

ABSTRACT

Objective To investigate the therapeutic effect of SPK1 gene transfected adipose derived mesenchymal stem cells(ADMSC)on experimental autoimmune encephalomyelitis mice and the effect on T helper cell 17(Th17)/regulatory T(Treg) cells balance. Methods EAE was induced by myelin oligodendrocyte glycoprotein 35-55 in mice.Totally 44 mice were randomly divided into four groups:normal control group(NC group),model group(EAE group),ADMSC group,and ADMSC-SPK1 group.Forty days after injection,the pathological changes of brain and spinal cord,Th17/Treg-related inflammatory markers in brain tissue,expressions of interleukin-17A(IL-17A)and forkhead box protein p3(Foxp3)in brain and spinal cord tissue,and flow cytometric results of spleen immune cells were detected. Results Forty days after the injection,serious inflammatory cell infiltration and demyelination occurred in the brain and spinal cord of EAE group,whereas demyelination and axonal injury were improved in ADMSC group and ADMSC-SPK1 group.Compared with EAE group,the ADMSC group and ADMSC-SPK1 group had significantly improved levels of IL-17A(


Subject(s)
Animals , Mice , Adipose Tissue/cytology , Cytokines , Encephalomyelitis, Autoimmune, Experimental/therapy , Interleukin-17 , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells/cytology , Mice, Inbred C57BL , Phosphotransferases (Alcohol Group Acceptor)/genetics , T-Lymphocytes, Regulatory/cytology , Th17 Cells/cytology , Transfection
4.
Braz. arch. biol. technol ; 63: e20190003, 2020. tab, graf
Article in English | LILACS | ID: biblio-1132227

ABSTRACT

Abstract Autologous fibrin matrices derived from the Leukocyte and Platelet Rich Plasma (L-PRP) and Leukocyte and Platelet Rich Fibrin (L-PRF) techniques present great potential to act as a bioactive scaffold in regenerative medicine, contributing to the maintenance of cell viability, proliferation stimulus and differentiation. In contrast, there are few studies that characterize the bioactive potential of these fibrin scaffolds by considering the process of production. The objective of this work was to characterize the intrinsic potential of maintaining cell viability of different fibrin scaffolds containing platelets and leukocytes. In order to achieve that, blood samples from a volunteer were collected and processed to obtain fibrin clots using the suggested techniques. To characterize the potential for in vitro viability, mesenchymal stem cells from human infrapatellar fat were used. The scaffolds were cellularized (1x105 cells/scaffolds) and maintained for 5 and 10 days under culture conditions with Dulbecco's Modified Eagle Medium, without addition of fetal bovine serum, and subsequently subjected to analyses by Fourrier transform infra-red spectroscopy, circular dichroism and fluorescence microscopy. The results demonstrated distinct intrinsic potential viability between the scaffolds, and L-PRP was responsible for promoting higher levels of viability in both periods of analysis. No viable cells were identified in the fibrin matrix used as controls. These results allow us to conclude that both fibrin substrates have presented intrinsic potential for maintaining cell viability, with superior potential exhibited by L-PRP scaffold, and represent promising alternatives for use as bioactive supports in musculoskeletal regenerative medicine.


Subject(s)
Humans , Male , Adult , Adipose Tissue/cytology , Tissue Engineering/methods , Platelet-Rich Plasma/cytology , Mesenchymal Stem Cells/physiology , Platelet-Rich Fibrin/cytology , Cell Survival , Spectroscopy, Fourier Transform Infrared , Tissue Scaffolds , Flow Cytometry
5.
Braz. arch. biol. technol ; 63: e20190260, 2020. graf
Article in English | LILACS | ID: biblio-1132251

ABSTRACT

Abstract Creating experimental models for obtaining stem cells from adipose tissue is necessary to elucidate their peculiar features. Objective: This study proposed a reliable reproducible and consistent experimental model for obtaining mesenchymal stem cells from adipose tissue. Material and Method: Lines of New Zealand rabbits, Wistar rats and CaviaPorcellus guinea pigs (4 animals per species) were used. Fatty tissue mesenchymal stem cells were removed from dorsal, epididymal and inguinal regions. Percentage viable cells and percentage cells expanded and submitted to chondrogenic differentiation were compared by animal species and collection site. Results: Chondrogenic differentiation occurred in a similar manner across all samples, independently of animal species or collection site. Among samples assessed, the inguinal region of rats yielded the highest percentage of viable and expanded cells. Conclusion: A reliable, reproducible and consistent model for obtaining mesenchymal stem cells was produced. Of the several variables analysed, the best results were obtained from the inguinal region of the rat.


Subject(s)
Animals , Guinea Pigs , Rats , Adipose Tissue/cytology , Mesenchymal Stem Cells/cytology , Rats, Wistar , Models, Animal
6.
Rev. bras. oftalmol ; 78(6): 355-363, nov.-dez. 2019. graf
Article in Portuguese | LILACS | ID: biblio-1057917

ABSTRACT

Resumo Objetivo: Verificar a presença das células-tronco mesenquimais (MSC) na área próxima ao nervo óptico de coelhos previamente lesado com álcool absoluto. Métodos: Os 12 coelhos da raça Nova Zelândia foram distribuídos em 2 lotes. Após sedação, cada olho do animal recebeu uma injeção retrobulbar de 1 ml de álcool absoluto em um dos olhos e de 1 ml de solução fisiológica 0,9% (SF) no olho contralateral. Após 15 dias deste procedimento inicial todos os olhos dos animais pertencentes ao lote A, receberam via retrobulbar, uma solução contendo MSC de tecido adiposo humano e previamente marcadas com Qdots,. Todos os olhos dos animais do lote B receberam solução PBS. Resultados: Após 15 dias desta última aplicação os animais foram sacrificados e as lâminas foram analisadas. A presença das MSC foi observada em 100% dos olhos dos animais do lote A. Conclusão: Os resultados sugerem que a marcação prévia das MSC com Qdots permitiu o acompanhamento das mesmas na região aplicada e em áreas mais internas do nervo óptico. A permanência de MSC após 15 dias de aplicação ao redor do nervo óptico sugere a viabilidade e possível participação das mesmas no processo de regeneração do tecido lesado. Nas condições deste estudo, a via de aplicação retrobulbar permitiu a mobilização das células tronco do local de aplicação até áreas centrais dos nervos ópticos nos animais do lote A, sugerindo que esta poderá ser uma via de acesso eficaz para as MSC no processo de regeneração de neuropatias ópticas.


Abstract Obtective: To verify the presence of mesenchymal stem cells (MSC) in the area close to the optic nerve of previously injured with absolute alcohol. Methods: Twelve New Zealand breed rabbits were divided into two groups, and after sedation, each eye of the animal received a retrobulbar injection of 1 ml of absolute ethanol in one eye, and 1 ml of physiological solution 0.9 % (PS) in the contralateral eye. After 15 days all eyes of animals belonging to group A, received via retrobulbar a solution containing MSCs from human adipose tissue (AT) and previously marked with Qdots, while all eyes of animals from group B received solution containing PBS. Results: The presence of MSC was observed in 100% of the eyes of the animals of group A and the more central areas near and into the optic nerve. Conclusion: The results suggest that the appointment of MSC with Qdots allowed their follow-up applied in the region and in the inner areas of the optic nerve. The MSC permanence after 15 days of application around the optic nerve suggests the feasibility and possible involvement of the same during the damaged tissue regeneration process. Under the conditions of this study, the route of retrobulbar application and the presence of the stem cells to the central areas of the optic nerves in animals of group A, suggests that this might be an effective approach for MSCs in regeneration process of optic neuropathies.


Subject(s)
Animals , Female , Rabbits , Optic Nerve Diseases/therapy , Adipose Tissue/cytology , Adipocytes , Mesenchymal Stem Cell Transplantation/methods , Mesenchymal Stem Cells/cytology , Nerve Regeneration , Optic Nerve/cytology , Semiconductors , Cell Differentiation , Cells, Cultured , Optic Nerve Diseases/chemically induced , Double-Blind Method , Quantum Dots , Injections, Intraocular
7.
Arq. bras. med. vet. zootec. (Online) ; 71(5): 1571-1581, set.-out. 2019. graf, ilus
Article in English | VETINDEX, LILACS | ID: biblio-1038673

ABSTRACT

There is a growing interest in the study of unspecialized mesenchymal stem cells, for there are still some discussions about their in vitro behavior. Regenerative medicine is a science undergoing improvement which develops treatments as cell therapy using somatic stem cells. In several studies, adipose tissue is presented as a source of multipotent adult cells that has several advantages over other tissue sources. This study aimed to characterize and evaluate the tagging of mesenchymal stem cells from the agoutis adipose tissue (Dasyprocta prymonolopha), with fluorescent intracytoplasmic nanocrystals. Fibroblast cells were observed, plastic adherent, with extended self-renewal, ability to form colonies, multipotency by differentiation into three lineages, population CD90 + and CD45 - expression, which issued high red fluorescence after the tagging with fluorescent nanocrystals by different paths and cryopreserved for future use. It is possible to conclude that mesenchymal stem cells from agouti adipose tissue have biological characteristics and in vitro behavior that demonstrate its potential for use in clinical tests.(AU)


Há um interesse crescente no estudo das células estaminais mesenquimais, não especializadas, pois ainda existem algumas discussões sobre seu comportamento in vitro. A medicina regenerativa é uma ciência em fase de crescimento que desenvolve tratamentos como terapia celular utilizando células estaminais somáticas. Em vários estudos, o tecido adiposo é apresentado como uma fonte de células adultas multipotentes que tem várias vantagens em relação a outras fontes de tecido. Este estudo teve como objetivo caracterizar e avaliar a marcação de células estaminais mesenquimais do tecido adiposo de cutias (Dasyprocta prymnolopha) com nanocristais intracitoplasmáticos fluorescentes. Observaram-se células fibroblásticas, aderentes ao plástico, com autorrenovação prolongada, capacidade de formar colônias, diferenciação em três linhagens, população CD90 + e expressão CD45, que emitiram alta fluorescência vermelha após a marcação com nanocristais fluorescentes por diferentes vias, e criopreservadas para uso futuro. É possível concluir que as células estaminais mesenquimais do tecido adiposo de cutias têm características biológicas e comportamentos in vitro que demonstram seu potencial para uso em testes clínicos.(AU)


Subject(s)
Animals , Adipose Tissue/cytology , Immunophenotyping/veterinary , Regenerative Medicine/methods , Nanoparticles , Mesenchymal Stem Cells , Dasyproctidae/genetics
8.
ABCD (São Paulo, Impr.) ; 32(4): e1465, 2019. graf
Article in English | LILACS | ID: biblio-1054591

ABSTRACT

ABSTRACT Background: Crohn's disease is a pathological condition that has different options of treatment, but there are patients who need other therapeutic approach, such as the use of adipose-derived mesenchymal stem cells. Aim: Systematic literature review to determine the different ways of adipose-derived mesenchymal stem cells administration in humans with luminal refractory and perianal fistulizing Crohn's disease. Methods: It was conducted a search for articles (from 2008 to 2018) on PubMed and ScienceDirect databases using the keywords Crohn's disease, fistulizing Crohn's disease, luminal Crohn's disease and transplantation of mesenchymal stem cells or mesenchymal stem cells or stromal cells. Thirteen publications were selected for analysis. Results: Only one study referred to the luminal Crohn´s disease. The number of cells administered was variable, occurring mainly through subcutaneous adipose tissue by liposuction. It could be highlighted the autologous transplant with exclusive infusion of mesenchymal stem cells. The procedures involved in pre-transplant were mainly curettage, setons placement and stitching with absorbable suture, and conducting tests and drug treatment for luminal Crohn´s disease. During transplant, the injection of mesenchymal stem cells across the fistula path during the transplant was mainly on the intestinal tract wall. Conclusion: Although the use of mesenchymal stem cells is promising, the transplant on the luminal region should be more investigated. The injection of mesenchymal stem cells, exclusively, is more explored when compared to treatment with other products. The preparation of the fistulizing tract and the location of cell transplantation involve standardized health care in most studies.


RESUMO Racional: Há diferentes opções de tratamento para a doença de Crohn, porém, em alguns casos, há a necessidade de outras abordagens terapêuticas, como o uso de células-tronco mesenquimais derivadas do tecido adiposo. Objetivo: Revisar sistematicamente a literatura para determinar as diferentes formas de administração das células-tronco mesenquimais derivadas do tecido adiposo em seres humanos com doença de Crohn refratária luminal e fistulizante perianal. Método: Buscaram-se artigos publicados entre 2008 e 2018 nas bases de dados PubMed e ScienceDirect, pelos descritores: Crohn's disease, fistulizing Crohns disease, luminal Crohns disease e transplantation of mesenchymal stem cells ou mesenchymal stem cell ou stromal cells. Treze artigos foram selecionados. Resultados: Somente um trabalho se referiu à doença luminal. A quantidade de células administradas foi variável, obtendo-se principalmente do tecido adiposo subcutâneo por lipoaspiração. Destacou-se o transplante autólogo com a infusão exclusiva de células-tronco mesenquimais. Os procedimentos realizados no pré-transplante foram principalmente o de curetagem, colocação de setons e suturas com fio absorvível, e de exames e tratamento medicamentoso para a doença luminal. No transplante, ocorreu a injeção das células por todo o trajeto fistuloso, principalmente nas paredes do trato. Conclusão: Embora o uso de células-tronco mesenquimais seja promissor, o transplante na região luminal deve ser mais investigado. A injeção exclusiva de células-tronco mesenquimais é mais explorada quando comparada ao tratamento conjunto com outros produtos. A forma de preparo do trato fistuloso e o local de transplante envolvem cuidados médicos padronizados na maioria dos estudos.


Subject(s)
Humans , Crohn Disease/therapy , Adipose Tissue/cytology , Rectal Fistula/therapy , Mesenchymal Stem Cell Transplantation/methods , Crohn Disease/complications , Adipose Tissue/transplantation , Rectal Fistula/etiology
9.
Biol. Res ; 52: 54-54, 2019. ilus
Article in English | LILACS | ID: biblio-1505774

ABSTRACT

BACKGROUND: IcarisideII (ICAII) could promote the differentiation of adipose tissue-derived stem cells (ADSCs) to Schwann cells (SCs), leading to improvement of erectile function (EF) and providing a realistic therapeutic option for the treatment of erectile dysfunction (ED). However, the underlying molecular mechanisms of ADSCs and ICAII in this process remain largely unclear. METHODS: ADSCs were treated with different concentrations of ICAII. Cell proliferation was determined by MTT assay. qRT-PCR and western blot were performed to detect expressions of SCs markers, signal transducer and activator of transcription-3 (STAT3), and microRNA-let-7i (let-7i). Luciferase reporter assay was conducted to verify the regulatory relationship between let-7i and STAT3. The detection of intracavernosal pressure (ICP) and the ratio of ICP/mean arterial pressure (MAP) were used to evaluate the EF in bilateral cavernous nerve injury (BCNI) rat models. RESULTS: ICAII promoted cell proliferation of ADSCs in a dose-dependent manner. The mRNA and protein levels of SCs markers were increased by ICAII treatment in a dose-dependent manner in ADSCs. Moreover, let-7i was significantly decreased in ICAII-treated ADSCs and upregulation of let-7i attenuated ICAII-induced promotion of SCs markers. In addition, STAT3 was a direct target of let-7i and upregulated in ICAII-treated ADSCs. Interestingly, overexpression of STAT3 abated the let-7i-mediated inhibition effect on differentiation of ADSCs to SCs and rescued the ICAII-mediated promotion effect on it. Besides, combination treatment of ADSCs and ICAII preserved the EF of BCNI rat models, which was undermined by let-7i overexpression. CONCLUSION: ICAII was effective for preserving EF by promoting the differentiation of ADSCs to SCs via modulating let-7i/STAT3 pathway.


Subject(s)
Animals , Male , Rats , Schwann Cells/drug effects , Flavonoids/pharmacology , Cell Differentiation/drug effects , Adipose Tissue/cytology , Mesenchymal Stem Cells/drug effects , Erectile Dysfunction/drug therapy , Transfection , Blotting, Western , Rats, Sprague-Dawley , Disease Models, Animal
10.
Acta cir. bras ; 33(11): 1016-1026, Nov. 2018. graf
Article in English | LILACS | ID: biblio-973481

ABSTRACT

Abstract Purpose: To evaluate renal repair in rats who had renal infarction induced by the obstruction of blood flow in the renal artery and were treated with transplantation of adipose tissue derived mesenchymal stem cell Methods: 16-week-old Wistar rats (n=72) were used, submitted to celiotomy and had of the renal artery and vein clipped for 24 hours. The animals were randomly assigned to 10 experimental homogeneous groups, corresponding to the treatments with phosphate-buffered saline (PBS) or adipose tissue derived mesenchymal stem cell (ADSC), duration of application (24 or 48 hours), and site of transplantation (lateral vein of the tail or intrarenal). After the treatments were performed, at 8 and 31 days, four animals in each group were subjected to left nephrectomy for histological studies. Results: Histologically, a higher amount of cell debris and tubules devoid of the epithelium and a higher degree of necrosis were observed in the groups treated with PBS, as opposed to a low degree of necrosis and higher tubular vascularization in the groups treated with ADSC, particularly in the group treated with intrarenal ADSC 48 hours after injury. Conclusion: The transplantation of ADSC positively contributed to the replacement of necrotic tissue by renal tubular cells, vascularization of the renal parenchyma, and restoration of the organ function.


Subject(s)
Animals , Male , Reperfusion Injury/surgery , Adipose Tissue/cytology , Acute Kidney Injury/surgery , Kidney/blood supply , Rats, Inbred Lew , Renal Artery Obstruction/surgery , Time Factors , Reperfusion Injury/pathology , Random Allocation , Reproducibility of Results , Treatment Outcome , Ultrasonography, Doppler, Color , Mesenchymal Stem Cell Transplantation/methods , Acute Kidney Injury/pathology , Kidney/pathology , Necrosis
11.
J. coloproctol. (Rio J., Impr.) ; 38(3): 240-245, July-Sept. 2018.
Article in English | LILACS | ID: biblio-954604

ABSTRACT

ABSTRACT Crohn's disease has an ever-increasing prevalence and incidence, with about 20% of patients developing perianal fistula with significant impact on their quality of life. Despite the medical and surgical treatments currently used, Crohn's-related fistula treatment continues to pose a challenge due to the low rates of efficacy associated with high recurrence rates. Recent clinical trials have shown promising results regarding safety and efficacy of local treatment of this condition with the use of adipose tissue-derived mesenchymal stem cells. Besides being pluripotent and poorly immunogenic, they have immunomodulatory and anti-inflammatory properties, which combined, may accelerate healing. Our main objective is to summarize the clinical trials we found, highlighting the efficacy rates of this therapy and the main limitations we found in the analysis of the results. We conclude that, in perianal fistulas refractory to conventional therapies, the treatment with adipose tissue-derived mesenchymal cells is safe with promising results that may change the current paradigm of Crohn's related fistula treatment.


RESUMO A incidência e a prevalência da doença de Crohn têm aumentado e, ao longo do decurso da doença, cerca de 20% dos doentes irão desenvolver fístulas perianais com impacto significativo na sua qualidade de vida. Apesar dos tratamentos médicos e cirúrgicos utilizados atualmente, o tratamento destas fístulas continua a constituir um desafio com baixas taxas de eficácia e com elevadas taxas de recorrência. Ensaios clínicos recentes têm demonstrado resultados promissores em termos de segurança e eficácia de tratamentos locais destas fístulas com o recurso a células estaminais mesenquimatosas derivadas do tecido adiposo que, além de pluripotentes e pouco imunogênicas, têm capacidades imunomoduladoras e anti-inflamatórias capazes de promover o processo de cicatrização. O objetivo desta revisão sistemática é sumarizar os ensaios clínicos encontrados, realçando as taxas de eficácia desta terapêutica e as principais limitações na análise dos resultados. Concluímos que, nas fístulas perianais refratárias ás terapias convencionais, o tratamento com com celulas estaminais mesenquimatosas derivadas do tecido adiposo é seguro e com resultados promissores que podem mudar o paradigma atual do tratamento das fistulas complexas associadas à Doença de Crohn.


Subject(s)
Humans , Male , Female , Crohn Disease/pathology , Rectal Fistula/therapy , Mesenchymal Stem Cells/cytology , Adipose Tissue/cytology , Treatment Outcome
12.
Acta cir. bras ; 33(5): 431-438, May 2018. tab, graf
Article in English | LILACS | ID: biblio-949342

ABSTRACT

Abstract Purpose: To evaluate the effects of this thymol-rich oil in the proliferation of human adipose tissue-derived stem cells. Methods: Stem cells were isolated from human adipose tissue by liposuction. After the first passage, cells were cultivated in triplicate for three days in control medium and medium supplemented with three oil samples (1.0 μg/mL, 5.0 μg/mL, and 25.0 μg/mL). Cells were analyzed by the MTT assay at passage 1 (P1), and cell proliferation of control and 1 μg/mL groups was determined with a hemocytometer at P2 and P3. Results: Viability of the essential oil-treated cells was significantly higher than the control group at P1 (p = 0.0008). The treatment with the oil, at a concentration of 1 µg/mL, led to increases of 24.8% at P1 and 43.0% at P3 in the rate of cell proliferation compared with control cells. Conclusion: Supplementing culture medium with essential oil of Lippia origanoides increased cell proliferation, especially at later passages.


Subject(s)
Humans , Adult , Thymol/pharmacology , Plant Oils/pharmacology , Oils, Volatile/pharmacology , Lippia/chemistry , Cell Proliferation/drug effects , Antioxidants/pharmacology , Stem Cells/drug effects , Plant Oils/chemistry , Oils, Volatile/chemistry , Lipectomy , Adipose Tissue/cytology , Culture Media
13.
Trends psychiatry psychother. (Impr.) ; 39(3): 196-201, July-Sept. 2017. tab, graf
Article in English | LILACS | ID: biblio-904580

ABSTRACT

Abstract Introduction The rationale of mesenchymal stem cells (MSCs) as a novel therapeutic approach in certain neurodegenerative diseases is based on their ability to promote neurogenesis. Hippocampal atrophy has been related to bipolar disorder (BD) in preclinical, imaging and postmortem studies. Therefore, the development of new strategies to stimulate the neurogenesis process in BD is crucial. Objectives To investigate the behavioral and neurochemical changes induced by transplantation of MSCs in a model of mania-like behavior induced by lisdexamfetamine dimesylate (LDX). Methods Wistar rats (n=65) received one oral daily dose of LDX (10 mg/kg) or saline for 14 days. On the 8th day of treatment, the animals additionally received intrahippocampal saline or MSC (1 µL containing 25,000 cells) or lithium (47.5 mg/kg) as an internal experimental control. Two hours after the last administration, behavioral and neurochemical analyses were performed. Results LDX-treated rats had increased locomotor activity compared to saline-saline rats (p=0.004), and lithium reversed LDX-related hyperactive behavior (p<0.001). In contrast, the administration of MSCs did not change hyperlocomotion, indicating no effects of this treatment on LDX-treated rats (p=0.979). We did not find differences between groups in BDNF levels (p>0.05) in the hippocampus of rats. Conclusion Even though these results suggest that a single intrahippocampal injection of MSCs was not helpful to treat hyperactivity induced by LDX and neither influenced BDNF secretion, we cannot rule out the possible therapeutic effects of MSCs. Further research is required to determine direct effects of LDX on brain structures as well as in other pathophysiological targets related to BD.


Resumo Introdução Células-tronco mesenquimais (CTMs) têm emergido como um promissor tratamento em diversas doenças neurodegenerativas devido a sua plasticidade e capacidade de regenerar tecidos. Estudos pré-clínicos, clínicos e de neuroimagem têm demonstrado a presença de atrofia hipocampal no transtorno bipolar (TB). Portanto, o desenvolvimento de tratamentos capazes de regenerar tecido lesado e estimular a neurogênese poderia ser útil. Objetivos Investigar mudanças comportamentais e neuroquímicas induzidas pelo transplante de CTMs no hipocampo de ratos em um modelo animal de mania induzido por dimesilato de lisdexanfetamina (LDX). Métodos Ratos Wistar (n=65) receberam LDX (10 mg/kg) ou solução salina por via oral durante 14 dias. No oitavo dia, os animais foram transplantados com injeção de CTMs ou solução salina (1 µL contendo 25.000 células) ou lítio (47,5 mg/kg) como controle interno do experimento. Duas horas após a última dose, foram realizadas análises comportamentais e neuroquímicas. Resultados Animais que receberam LDX tiveram um aumento da atividade locomotora comparados ao grupo que recebeu solução salina (p=0,004); já o lítio reverteu a hiperatividade locomotora desses animais (p<0,001). Os animais que receberam CTMs não apresentaram alterações no comportamento, indicando ausência de efeitos sobre hiperatividade locomotora. Os níveis de BDNF hipocampais não diferiram entre os grupos (p>0.05). Conclusão Não foi possível demonstrar efeitos neuroprotetores das CTMs, administradas em dose única, em um modelo animal de mania induzido por LDX. No entanto, não se pode descartar os possíveis efeitos terapêuticos das CTMs. Mais estudos são necessários para determinar os efeitos das CTMs em estruturas cerebrais e outros alvos fisiopatológicos relacionados ao TB.


Subject(s)
Animals , Male , Bipolar Disorder/therapy , Mesenchymal Stem Cell Transplantation , Bipolar Disorder/metabolism , Cells, Cultured , Adipose Tissue/cytology , Rats, Wistar , Lithium Compounds/pharmacology , Antimanic Agents/pharmacology , Brain-Derived Neurotrophic Factor/metabolism , Disease Models, Animal , Lisdexamfetamine Dimesylate , Proof of Concept Study , Hippocampus/surgery , Hippocampus/metabolism , Mice, Inbred C57BL , Motor Activity/drug effects , Motor Activity/physiology
14.
J. appl. oral sci ; 25(1): 10-19, Jan.-Feb. 2017. tab, graf
Article in English | LILACS, BBO | ID: biblio-841166

ABSTRACT

Abstract Objectives This study aimed to evaluate the potential of adipose-derived stem cells (ASCs) combined with a modified α-tricalcium phosphate (α-TCP) or gelatin sponge (GS) scaffolds for bone healing in a rat model. Material and Methods Bone defects were surgically created in the femur of adult SHR rats and filled with the scaffolds, empty or combined with ASCs. The results were analyzed by histology and histomorphometry on days seven, 14, 30, and 60. Results Significantly increased bone repair was observed on days seven and 60 in animals treated with α-TCP/ASCs, and on day 14 in the group treated with GS/ASCs, when compared with the groups treated with the biomaterials alone. Intense fibroplasia was observed in the group treated with GS alone, on days 14 and 30. Conclusions Our results showed that the use of ASCs combined with α-TCP or GS scaffolds resulted in increased bone repair. The higher efficacy of the α-TCP scaffold suggests osteoconductive property that results in a biological support to the cells, whereas the GS scaffold functions just as a carrier. These results confirm the potential of ASCs in accelerating bone repair in in vivo experimental rat models. These results suggest a new alternative for treating bone defects.


Subject(s)
Animals , Male , Biocompatible Materials/pharmacology , Bone Regeneration/drug effects , Calcium Phosphates/pharmacology , Adipose Tissue/cytology , Stem Cell Transplantation/methods , Tissue Scaffolds , Gelatin Sponge, Absorbable/pharmacology , Osteogenesis/drug effects , Rats, Inbred SHR , Tetrazolium Salts , Time Factors , Wound Healing/drug effects , Biocompatible Materials/therapeutic use , Calcium Phosphates/therapeutic use , Cell Adhesion/drug effects , Cells, Cultured , Reproducibility of Results , Treatment Outcome , Models, Animal , Cell Proliferation/drug effects , Femur/surgery , Femur/pathology , Fibroblasts/drug effects , Formazans , Gelatin Sponge, Absorbable/therapeutic use
15.
Cell Journal [Yakhteh]. 2017; 19 (1): 117-126
in English | IMEMR | ID: emr-185798

ABSTRACT

Objective: Sulfur mustard [SM] is a potent mutagenic agent that targets several organs, particularly lung tissue. Changes in morphological structure of the airway system are associated with chronic obstructive pulmonary deficiency following exposure to SM. Although numerous studies have demonstrated pathological effects of SM on respiratory organs, unfortunately there is no effective treatment to inhibit further respiratory injuries or induce repair in these patients. Due to the extensive progress and achievements in stem cell therapy, we have aimed to evaluate safety and potential efficacy of systemic mesenchymal stem cell [MSC] administration on a SM-exposed patient with chronic lung injuries


Materials and Methods: In this clinical trial study, our patient received 100x106 cells every 20 days for 4 injections over a 2-month period. After each injection we evaluated the safety, pulmonary function tests [PFT], chronic obstructive pulmonary disease [COPD] Assessment Test [CAT], St. George's Respiratory Questionnaire [SGRQ], Borg Scale Dyspnea Assessment [BSDA], and 6 Minute Walk Test [6MWT]. One-way ANOVA test was used in this study which was not significant [P>0.05]


Results: There were no infusion toxicities or serious adverse events caused by MSC administration. Although there was no significant difference in PFTs, we found a significant improvement for 6MWT, as well as BSDA, SGRQ, and CAT scores after each injection


Conclusion: Systemic MSC administration appears to be safe in SM-exposed patients with moderate to severe injuries and provides a basis for subsequent cell therapy investigations in other patients with this disorder


Subject(s)
Humans , Mustard Gas/adverse effects , Adipose Tissue/cytology , Mesenchymal Stem Cells , Long Term Adverse Effects , Iran
16.
Cell Journal [Yakhteh]. 2017; 19 (1): 137-145
in English | IMEMR | ID: emr-185800

ABSTRACT

Objective: Adipose derived stem cells [ASCs], as one of the important stromal cells in the tumor microenvironment, are determined with immunomodulatory effects. The principle aim of this study was to evaluate the immunosuppressive effects of ASCs on natural killer [NK] cells


Materials and Methods: In this experimental study, we assessed the expressions of indolamine 2, 3-dioxygenase [IDO1], IDO2 and human leukocyte antigen-G5 [HLA-G5] in ASCs isolated from breast cancer patients with different stages as well as normal individuals, using quantitative reverse transcriptase-polymerase chain reaction [qRT-PCR]. Immunomodulatory effects of ASCs on the expression of CD16, CD56, CD69, NKG2D, NKp30, NKG2A and NKp44 was also assessed in peripheral blood lymphocytes [PBLs] by flow-cytometry


Results: Our result showed that IDO1, IDO2 and HLA-G5 had higher mRNA expressions in ASCs isolated from breast cancer patients than those from normal individuals [P>0.05]. mRNA expression of these molecules were higher in ASCs isolated from breast cancer patients with stage III tumors than those with stage II. The indirect culture of ASCs isolated from breast cancer patients and normal individuals with activated PBLs significantly reduced NKG2D+ and CD69+ NK cells [P<0.05]


Conclusion: Results of the present study suggest more evidences for the immunosuppression of ASCs on NK cells, providing conditions in favor of tumor immune evasion


Subject(s)
Adult , Female , Humans , Middle Aged , Adipose Tissue/cytology , Mesenchymal Stem Cells/immunology , Killer Cells, Natural/immunology , Immunomodulation , Immunosuppression Therapy , Iran
17.
Acta cir. bras ; 31(12): 821-825, Dec. 2016. tab, graf
Article in English | LILACS | ID: biblio-837663

ABSTRACT

ABSTRACT PURPOSE: To evaluate the Adipose Stem Cells (ACS) therapy efficacy on the time and quality of wound healing process in rats. METHODS: Nine male Wistar rats were randomly distributed into three groups I) 7 days of healing; II) 14 days of healing; III) 21 days of healing. Four incisions were made on the dorsal surface of each rat and then treated with intralesional ACS, meloxicam, and no treatment and ACS+meloxicam. Macroscopic evaluation was measured by percentage of healing and histopathological by hematoxylin-eosin was performed. RESULTS: All groups have the wound reduced during the three weeks (p<0.001) and after 14 days of healing had greater reduction than others. Wounds treated with ASC had accelerated healing in relation to no treatment and only meloxicam (p<0.001), excepting the ASC+Meloxicam that was similar (p=0.13). There was no difference in histopathological analysis between lesions. CONCLUSION: Adipose stem cell have benefits in reducing time of healing of experimental model of wound in rats, observed 7 days of after application.


Subject(s)
Animals , Male , Rats , Stem Cells/physiology , Wound Healing/physiology , Wounds and Injuries/therapy , Adipose Tissue/cytology , Stem Cell Transplantation , Treatment Outcome , Rats, Wistar , Disease Models, Animal , Antigens, Surface
18.
Acta cir. bras ; 31(9): 578-585, Sept. 2016. tab, graf
Article in English | LILACS | ID: lil-795994

ABSTRACT

ABSTRACT PURPOSE: To analyze the healing effects of stromal vascular fraction (SVF) application compared to wound dressing with 2% silver sulfadiazine in full thickness burn wounds in rats. METHODS: Animals were divided into two groups: 2% silver sulfadiazine group and SVF group. Both groups received occlusive bandages while the first one was treated with 2% silver sulfadiazine and the latter was treated with injections of SVF prepared from adipose tissue extracted from an animal donor. The animals were accompanied through 3, 7 and 30 days for evaluation of macroscopic, microscopic and morphometric aspects. RESULTS: On day three, a significant increase (p<0.05) of infiltration of polymorphonuclear, fibrin formation and fibroblasts migration in SVF group was observed. On the 7th day the mononuclear infiltrate, angiogenesis, collagen and fibroblasts were significantly increased in the SVF group (p<0.05). At 30 days significantly increased collagen deposition was observed in the SVF group (p<0.05) . CONCLUSION: Adipose tissue derived stromal vascular fraction injections promotes better wound repair than 2% silver sulfadiazine in the treatment of full thickness burn in rats during the evaluated experimental period.


Subject(s)
Animals , Male , Silver Sulfadiazine/administration & dosage , Wound Healing , Burns/therapy , Adipose Tissue/transplantation , Anti-Infective Agents, Local/administration & dosage , Bandages , Wound Healing/drug effects , Burns/surgery , Burns/pathology , Adipose Tissue/cytology , Stromal Cells/cytology , Stromal Cells/transplantation , Rats, Wistar , Disease Models, Animal , Microscopy
19.
Rev. chil. cir ; 68(4): 295-301, jul. 2016. graf, tab
Article in Spanish | LILACS | ID: lil-788897

ABSTRACT

Objetivo Obtener células estromales derivadas del tejido adiposo, medir y comparar las tasas de viabilidad antes e inmediatamente después un ciclo de criopreservación con diferentes combinaciones de criopreservantes de manera de obtener el mejor medio de criopreservación. Material y método Medición de la tasa de viabilidad poscriopreservación de células estromales derivadas del tejido adiposo obtenidas de 5 pacientes utilizando medios definidos (DMEM/Ham F12) libres de suero bovino y suplementados con una de los siguientes combinaciones de compuestos: dimetilsulfóxido (DMSO) 10%; DMSO 10% + trehalosa 7,6%; DMSO 10% + albúmina humana 10% y DMSO 10% + trehalosa 7,6% + albúmina humana 10%, mediante citometría de flujo con ioduro de propidio. Resultados No existen diferencias estadísticamente significativas en las tasas de viabilidad de las células estromales posterior a un ciclo de criopreservación. Sin embargo, se observa una tendencia a mejorar la tasa de recuperación de células vitales al agregar albúmina humana. Conclusiones No se observaron diferencias significativas entre las condiciones estudiadas, sugiriendo que ninguna es superior a las demás en cuanto a rendimiento. Es así como podemos afirmar que la criopreservación de las células estromales derivadas del tejido adiposo en un medio que combine DMEM/F12 con DMSO 10% + trehalosa 7,6% + albúmina humana 10% no logra una tasa de recuperación de células vitales significativamente mayor que las congeladas solo con DMSO 10%.


Aim To obtain stromal cells derived from adipose tissue, to measure and compare viability rates before and immediately after cryopreservation cycle, using different combinations of cryoprotective agents in order to identify the best cryopreservation medium. Material and method Viability rate after cryopreservation of stromal cells derived from adipose tissue were assessed by flow cytometry with propidium iodide. Samples of stromal cells obtained from 5 patients were kept defined, bovine serum-free media (DMEM/Ham-F12), supplemented with one of the following combinations of compounds: 10% dymethylsulfoxide (DMSO); Trehalose 10% DMSO + 7.6%; 10% DMSO + 10% human albumin and 10% DMSO + 7.6% Trehalose + 10% human albumin. Results No statistically significant differences were observed in the viability rates of stromal cells derived from adipose tissue after a cryopreservation cycle. However, we observed a tendency towards improvement of recovery rate when human albumin was added to the medium. Conclusions None of the studied conditions proved superior to others in terms of cell vitality after a cryopreservation cycle. Hence, we conclude that the cryopreservation of stromal cells derived from adipose tissue in an environment that combines DMEM/F12 with 10% DMSO + 7.6% Trehalose + human albumin 10% does not achieve a significantly higher recovery rate than only frozen solely with DMSO 10%.


Subject(s)
Humans , Cryopreservation/methods , Cell Survival/drug effects , Stromal Cells/physiology , Cryoprotective Agents/pharmacology , Trehalose/pharmacology , Dimethyl Sulfoxide/pharmacology , Adipose Tissue/cytology , Serum Albumin, Human/pharmacology , Freezing
20.
Pesqui. vet. bras ; 36(supl.1): 21-32, June 2016. tab, graf, ilus
Article in Portuguese | LILACS, VETINDEX | ID: lil-798019

ABSTRACT

O objetivo deste estudo foi comparar o potencial osteogênico das células tronco mesenquimais extraídas da medula óssea (CTM-MO) com as do tecido adiposo (CTM-AD) de cães adultos. As células foram caracterizadas fenotipicamente quanto à expressão de CD29, CD90, CD34 e CD45 e submetidas à diferenciação adipogênica e condrogênica por 21 dias e osteogênica por 7, 14 e 21 dias. Foram constituídos quatro grupos: 1) CTM-MO em meio osteogênico, 2) CTM-MO em meio basal, 3) CTM-AD em meio osteogênico e 4) CTM-AD em meio basal. Aos 7, 14 e 21 dias de diferenciação osteogênica as culturas foram submetidas às avaliações da conversão de MTT em formazan, da atividade da fosfatase alcalina (FA), da síntese de colágeno e de matriz mineralizada, avaliação do número de células por campo e foram quantificados os transcritos gênicos para osterix, sialoproteina óssea (BSP), osteonectina (ON) e osteocalcina (OC). Tanto as células extraídas da medula óssea quanto do tecido adiposo mostraram elevada expressão de marcadores para células tronco e baixa expressão de marcadores de células hematopoiéticas (menor que 2%). Além disso, foram capazes de se diferenciar em osteoblastos, condrócitos e adipócitos. As CTM-AD submetidas à diferenciação osteogênica mostraram maior conversão do MTT em formazan que as CTM-MO, sob mesmas condições aos 7 e 21 dias. O número de células por campo, a atividade da FA, a síntese de colágeno e de matriz mineralizada foram superior nas CTM-AD em diferenciação, em relação às CTM-MO sob as mesmas condições, em todos os tempos estudados. As expressões de osterix, BSP e OC foram predominantemente superiores nas CTM-MO diferenciadas, mas a expressão de ON foi superior nas CTM-AD diferenciadas aos 7, 14 e 21 dias. Conclui-se que as CTM-AD apresentam maior potencial osteogênico que as CTM-MO quando extraídas de cães adultos.(AU)


The aim of this study was to compare the osteogenic potential of mesenchymal stem cells obtained from bone marrow (BM-MSC) with those extracted from adipose tissue (AT-MSC) of adult dogs. The cells were phenotypically categorized according to the expression of CD29, CD90, CD34 and CD45, and submitted to adipogenic and chondrogenic differentiation for 21 days and osteogenic differentiation for 7, 14 and 21 days. Four groups were formed: BM-MSC in osteogenic medium (1), BM-MSC in basal medium (2), AT-MSC in osteogenic medium (3) and ATMSC in basal medium (4). On days 7, 14 and 21 of osteogenic differentiation, the cultures were submitted to evaluations of MTT conversion in formazan, of alkaline phosphatase activity (AP), of collagen and mineralized matrix synthesis, evaluation of the number of cells per field and there was quantification of the gene transcripts for osterix, bone sialoprotein (BSP), osteonectin (ON) and osteocalcin (OC). Both the cells obtained from bone marrow and those from adipose tissue showed high expression of stem cells markers and low expression of hematopoietic cells markers (lower than 2%). Besides, they were able to differentiate into osteoblasts, chondrocytes and adipocytes. AT-MSC submitted to osteogenic differentiation showed higher MTT conversion in formazan than BM-MSC, under the same conditions on days 7 and 21. The number of cells per field, the AP activity, the collagen and mineralized matrix synthesis were higher in AT-MSC en differentiation, in relation to BM-MSC under the same conditions in all evaluated times. Expressions of osterix, BSP and OC were predominantly higher in differentiated BMMSC, however the expression of ON was higher AT-MSC differentiated on days 7, 14 and 21. In conclusion, AT-MSC present higher osteogenic potential than BM-MSC when extracted from adult dogs.(AU)


Subject(s)
Animals , Dogs , Adipose Tissue/cytology , Bone Marrow Cells , Osteogenesis , Stem Cells , Bone Regeneration
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