O carcinoma espinocelular de cabeça e pescoço (CECP) é um problema de saúde pública que apresenta alta taxa de mortalidade, frequentemente relacionado à presença de recorrências locais e metástases. A descoberta de um pequeno subconjunto de células tumorais com características semelhantes às células-tronco, conhecidas como células-tronco tumorais (CTTs), tem sido relatadas como as principais responsáveis pelo início, progressão e recidiva do CECP. O processo de metástase nestas neoplasias é bastante complexo e envolve o desprendimento de células epiteliais tumorais do local de aparecimento primário devido à subexpressão ou superexpressão de algumas proteínas específicas nestas células, caracterizando um processo conhecido como transição epitélio-mesenquimal (EMT). A compreensão dos mecanismos envolvidos no processo de EMT têm sido investigados para o desenvolvimento de terapias específicas. O ácido retinoico (AR) vem sendo empregado em diversas terapias devido a sua capacidade de controlar a proliferação e promover diferenciação celular, entretanto, anormalidades na expressão ou função de seus receptores são relatadas em muitos tipos de células do câncer. Este estudo tem por objetivo correlacionar a expressão de marcadores do processo de EMT, marcador de célula tronco tumoral (ALDH1) e receptores do ácido retinoico e de retinoide X (isoformas α e β) em amostras teciduais provenientes de portadores de CECP primários, além, de correlacionar os resultados obtidos com os parâmetros clínicos, características histopatológicas e prognóstico destes pacientes em um período de acompanhamento de 7 anos. / The head and neck squamous cell carcinoma (HNSCC) is a public health problem that presents high mortality rates in relation to the presence of local recurrences and metastases. A finding of a small subset of tumor cells with stem like-cells characteristics, known as cancer stem cells (CTTs), has been reported as being primarily responsible for the onset, progression and recurrence of CECP. The metastasis process in these neoplasms is quite complex and involves the tumor epithelial cells detachment from the primary site of appearance due to underexpression or overexpression of some specific proteins in these cells, characterizing the epithelial-mesenchymal transition (EMT) process. An understanding of the mechanisms involved in the EMT process has been investigated for the development of specific therapies. Retinoic acid (AR) has been used in several therapies because its ability to control the proliferation and promote cell differentiation, however, abnormalities in the expression and function of its receptors are reported in many types of cancer cells. The aim of this study was to correlate the expression EMT process markers, tumor stem cell marker (ALDH1), retinoic acid and retinoic acid X receptors (α and β isoforms) in tissue samples from primary CECP, in addition, to correlate the results with the clinical parameters, histopathological and prognostic characteristics of these patients in a 7 years follow-up.

Colonic stromal cells provide critical structural support but also regulate immunity, tolerance and inflammatory responses in the mucosa. Substantial variability and plasticity of mucosal stromal cells has been reported but a paucity of distinct marker genes exist to identify distinct cell states. Here single-cell RNA-sequencing is used to document heterogeneity and subtype specific markers of individual colonic stromal cells in human and mouse. Marker-free transcriptional clustering of fibroblast-like cells derived from healthy human tissue reveals distinct populations corresponding to myofibroblasts and three transcriptionally and functionally dissimilar populations of fibroblasts. A SOX6 high fibroblast subset occupies a position adjacent to the epithelial basement membrane and expresses multiple epithelial morphogens including WNT5A and BMP2. Additional fibroblast subtypes show specific enrichment for chemokine signalling and prostaglandin E₂ synthesis respectively. In ulcerative colitis, substantial remodelling occurs with depletion of the SOX6 high population and emergence of an immune enriched population expressing genes associated with fibroblastic reticular cells including CCL19, CCL21 and IL33. A large murine dataset comprising over 7,000 colonic mesenchymal cells from an acute colitis model and matched healthy controls reveals strong preservation of the SOX6 high and myofibroblast transcriptional signatures. Unsupervised pseudotemporal ordering is used to relate fibroblast subsets to one another producing a branched developmental hierarchy that includes a potential progenitor population with mesothelial characteristics at its origin. This work provides a molecular basis for re-classification of colonic stromal cells and identifies pathological changes in these cells underpinning inflammation in UC.

Idiopathic pulmonary fibrosis (IPF) is a disease of unknown aetiology, characterised by the progressive and irreversible scarring of parenchymal lung tissue that leads to respiratory failure and death. The disease is understood to be driven by an impaired and aberrant wound healing response, with an inappropriate reactivation of developmental signalling. The greatest risk factor for the disease is age, which is a process intimately associated with an increase in the burden of senescent cells. Such cells acquire a unique secretory phenotype and are known to have a significant impact on their local micro-environment. It was hypothesised that an alteration in epithelial-mesenchymal secretory communication, due to senescent-like changes in the fibroblast phenotype, may detrimentally contribute to lung homeostasis. An in vitro model of the lung airway was established in which primary human lung fibroblasts (HLFs) were co-cultured with human bronchial epithelial cells (HBECs). HBECs were cultured on a semi-permeable, transwell insert and co-cultured with either normal (NHLF), fibrotic (FHLF) or senescent fibroblasts. Over 72 hrs of co-culture, wound healing was assessed, via an epithelial scratch assay, and epithelial regeneration was measured, via trans-epithelial electrical resistance. Co-culture with NHLFs improves epithelial regeneration, however, FHLFs and senescent cells in co-culture show a diminished ability to promote epithelial regeneration and wound repair. The secretory repertoire of these cells contains elevated levels of IL-6, CXCL8, CXCL1 and GCSF (when assessed at both an RNA and protein level), factors strongly associated with the senescent phenotype. Targeting this secretome via treatment with the JAK 1/2 inhibitor Ruxolitinib attenuates these impairments and may point towards a new therapeutic strategy for the treatment of IPF.

Made available in DSpace on 2014-06-11T19:32:22Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-03-17Bitstream added on 2014-06-13T19:43:01Z : No. of bitstreams: 1 penteado_fcl_dr_arafcf.pdf: 1008387 bytes, checksum: 40b22d48514b2c755f67d69d9ae8cff6 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Universidade Estadual Paulista (UNESP) / Alguns trabalhos realizados recentemente relatam que as células-tronco mesenquimais (CTM) podem ser induzidas à aquisição de marcadores hepatocíticos pelo transplante em modelos animais de dano hepático, ou pelo cultivo in vitro com fatores de crescimento e citocinas. O presente trabalho teve por objetivo avaliar o comportamento das CTM frente à indução da diferenciação hepatocítica. As CTM foram isoladas da medula óssea de quatro doadores saudáveis, caracterizadas e submetidas ao protocolo de indução à diferenciação hepatocítica in vitro e in vivo. As células induzidas in vitro apresentaram mudanças na sua morfologia, mostrando a morfologia semelhante à do hepatócito, porém, o perfil imunofenotípico não foi modificado. As células induzidas também não apresentaram o aumento dos transcritos de albumina, citoqueratina 18 e citoqueratina 19 quando analisadas por RT-PCR em tempo real, e não alteraram a expressão de albumina, citoqueratina 18 e alfafetoproteína como demonstrado por imunofluorescência. Quando analisadas in vivo, as CTM demonstraram o potencial migratório para o tecido hepático danificado de camundongos imunodeficientes. Em conjunto, os resultados sugerem que as CTM da medula óssea não são capazes de se diferenciar em hepatócitos quando estimuladas in vitro pela metodologia utilizado neste trabalho, mas são capazes de migrar para o tecido hepático danificado in vivo, o que sugere o seu papel no reparo do fígado. A contribuição para o reparo pode estar associada com o efeito parácrino dessas células. / Some recently works have been reported that mesenchymal stem cells (MSC) can be induced to the acquisition of hepatocytic markers for the transplant in animal models of liver damage, or for the in vitro culture with growth factors and cytokines. The present work aim is to evaluate the behavior of the MSC in front of the induction of the hepatocytic differentiation. The MSC was isolated from the bone morrow of 4 normal donators, characterized and submitted to the protocol of in vitro and in vivo induction of hepatocytic differentiation. The in vitro induced cells showed morphology changes acquiring hepatocytes-like morphology. However, the immunophenotypic profile of those cells was not modified. The induced cells did not present increase of the albumin, cytokeratin 18 and cytokeratin 19 transcripts, when analyzed by real time RTPCR. The expression of albumin, cytokeratin 18 and alpha foetoprotein was also not modified as demonstrated by immunofluorescence. In vivo, the MSC have demonstrated the migratory potential for the damaged liver of immunodeficient mice. Together, the results suggest that the bone morrow MSC are not capable of in vitro hepatocytic differentiating according to the approach in this work, but are capable to homming into damaged hepatic tissue in vivo. This migration capacity suggests their role in the repair mechanisms.

Regenerative medicine and cell therapies hold great potential to treat a variety of medical conditions. Product characterisation of these therapies is particularly difficult as they pose regulatory challenges due to donor heterogeneity and the lack of standardised lot release tests that can reliably predict in vivo function. Human mesenchymal stem cells (hMSCs), also called multipotent stem cells or mesenchymal stromal cells, are a viable option in cell therapies due to their immunosuppressive and pro-angiogenic functions. Currently there are no standardised methods or potency assays to quantify these properties. To address this, five individual hMSCs lines from different donors were created and characterised based upon growth rate, differentiation capability and extracellular surface protein expression. A novel multiparameter flow cytometry method to characterise the cells based upon extracellular surface markers was developed that supports high-throughput and high-content analyses. Three candidate lines were taken forward and assessed in multiple in vitro bioassays that examined the hMSC immunosuppressive response to a defined inflammatory environment, effect on T-cell proliferation, and effect on a mixed lymphocyte population. Next, the angiogenic properties were assessed using human umbilical vein endothelial cells (HUVECs) tube formation as a model for cardiac regeneration. This involved utilising automated time lapse microscopy techniques coupled with image analysis software to quantify endothelial to tube formation. Further analysis of the hMSC secretome revealed differences in the levels of pro-angiogenic cytokines such as vascular endothelial growth factor, hepatocyte growth factor and IL-8. Significant differences in angiogenic potency were found between the hMSC lines. This thesis highlights the need to develop specific assays that reflect the intended clinical action. Taken together, these quantitative approaches provide valuable tools to measure hMSC quality and potency, and supports continued efforts to improve characterisation strategies for cellular therapies.

Master of Science / Department of Clinical Sciences / Walter Renberg / Objective: To evaluate the safety and effect of a single simultaneous intra-articular and intravenous injection of autologous adipose-derived stromal vascular fraction (SVF) and platelet rich plasma (PRP) on coxofemoral osteoarthritis (OA) in dogs. Methods: This was a randomized, double-blind, placebo-controlled prospective pilot trial of simultaneous intra-articular and intravenous SVF and PRP for coxofemoral OA. Dogs with coxofemoral OA causing signs of lameness or discomfort were evaluated by orthopedic exam, visual lameness score, Canine Brief Pain Inventory (CBPI), goniometry, visual analogue scale (VAS), and pressure-sensitive walkway (PSW) at week 0 (baseline), and at 4, 8, 12 and 24 weeks after injection. Joint radiographs were scored at 0 and 24 weeks. Results: Twenty two client-owned dogs with naturally occurring OA of the coxofemoral joints were enrolled (12 placebo-control, 10 SVF-treated). CBPI pain severity scores were lower in the treatment group at 24 weeks compared to the placebo group (p=0.042). The VAS score for the treatment group was significantly greater at 0 weeks than at 4, 8, or 24 weeks (p<0.05). When dogs with low quartile baseline PVF (25th percentile) were compared, the treatment group had statistically higher PVF at all post-injection time points when compared to the placebo group. After SVF injection, fewer dogs in the treated group were lame compared to the control group. Clinical Significance: This study is the first to utilize objective data from PSW as an outcome measure for dogs treated with SVF and PRP for coxofemoral OA. No adverse events were noted. Improvements in some measured parameters in the treated dogs compared to those in the placebo group.

Made available in DSpace on 2014-06-11T19:27:56Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-02-28Bitstream added on 2014-06-13T20:57:18Z : No. of bitstreams: 1 salgado_bs_me_botfm.pdf: 482776 bytes, checksum: 23c491910cd4c7da0d422455a7e9b2df (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Transição epithelial-mesenquimal (EMT) é a conversão de células epiteliais polarizadas para células migratórias com fenótipo fibroblasto-símile. A EMT está envolvida na progressão e metástase em diversos cânceres nos seres humanos, porém permanece a ser mais bem explorada na literatura veterinária. O objetivo desta pesquisa foi avaliar a imunoexpressão de proteínas relacionadas à EMT nos carcinomas mamários de cadelas (CMCs). Seis proteínas foram avaliadas por meio de imunoistoquímica em 94 amostras de CMCs. Tecidos mamários não neoplásicos de 17 cadelas e amostras de 9 tumores mamários benignos de cadelas foram avaliados de modo a determinar o perfil de imunoexpressão de Snai-1. Características anatomopatológicas foram comparadas com a imunoexpressão de proteínas relacionadas à EMT nos CMCs. A perda de proteínas epiteliais e/ou a aquisição de proteínas mesenquimais foi observada principalmente em neoplasias com evidência de invasão estromal; entretanto, somente foi observada significância estatística quando comparado S100A4 e invasão vascular. Snai-1 foi observado em células luminais de neoplasias simples malignas e em células mioepiteliais de tumores benignos ou malignos de caráter complexo, sendo também significativamente relacionado à baixa de expressão de Caderina-E. Conclui-se que a perda de proteínas epiteliais e/ou a aquisição de proteínas mesenquimais está associada com EMT e pode possuir importante papel na avaliação de CMCs. O padrão único de imunoexpressão de Snai-1 pode ajudar a distinção entre um adenoma e um carcinoma não metastático e aparenta estar relacionado à conversão de células mioepiteliais a um fenótipo mesenquimal completo. A perda de Caderina-E e citoqueratina e a mudança no padrão de imunoexpressão de Snai-1, Caderina-N, S100A4 e MMP-2 indica a ocorrência de EMT em carcinomas mamários de cadelas... / Epithelial-mesenchymal transition (EMT) is defined as switching of polarized epithelial cells to a migratory fibroblastoid phenotype. EMT is known to be involved in the progression and metastasis of various cancers in humans, but this specific process is still little explored in the veterinary literature. The aim of this research was to evaluate the expression of EMT-related proteins in canine mammary carcinomas (CMCs). The expression of six EMT-related proteins in CMC of 94 female dogs was evaluated by immunohistochemistry. Additionally, mammary tissues from 17 female dogs with no history of mammary tumor development and from 9 bitches with benign tumors were evaluated in order to determine Snai-1 immunoexpression patterns. Anatomopathological characteristics were compared with the expression of EMTrelated proteins in CMCs. Loss of epithelial protein and/or acquisition of the expression of mesenchymal proteins were observed, particularly in tumors with evidence of stromal invasion; however, significance was only observed between the S100A4 and vascular invasion. Snai-1 was only expressed in luminal cells of histologically malignant tumors and in myoepithelial cells of benign and malignant complex tumors and was significantly related to E-cadherin loss. In conclusion, loss of epithelial proteins and/or the acquisition of mesenchymal proteins are associated with EMT and may have an important role in the evaluation of CMC patients. The unique immunoexpression pattern of Snai-1 could help to distinguish between an adenoma and a non-metastatic carcinoma and seems to be related to conversion of myoepithelial cells to a complete mesenchymal-like phenotype. Loss of E-cadherin and cytokeratin and change of immunoexpression pattern of Snai-1, N-cadherin, S100A4 and MMP-2 indicate the occurrence of EMT in canine mammary carcinomas and should result in an en bloc resection or a close follow-up.

Made available in DSpace on 2014-06-11T19:24:45Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-19Bitstream added on 2014-06-13T20:52:31Z : No. of bitstreams: 1 moroz_a_me_botfm.pdf: 608341 bytes, checksum: 715f3ed5332daf7487462c8fa4111838 (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Levando-se em consideração avanços tecnológicos na área médica e o impacto dos programas de saúde que determinaram curvas de longevidade cada vez maiores e taxas de natalidade cada vez menores, o novo desafio do gestor público são as conseqüências: o envelhecimento e a vida como uma “doença crônica”. Entre os principais desafios está a abordagem das doenças crônico-degenerativas que determinam o aumento de lesões cartilaginosas articulares. A débil capacidade de regeneração e as limitações das alternativas de tratamento fazem as técnicas derivadas da biotecnologia, como o transplante autólogo de condrócitos (TAC) e o uso de células-tronco, o foco das investigações. O TAC requer coleta de material cartilaginoso de área sadia, podendo causar nova lesão; no entanto, pode-se evitar este perigo com o uso de células-tronco. As células-tronco mesenquimais, adultas, podem se diferenciar em condrócitos mediante o uso de meio de cultura específico em consonância a um arcabouço 3D, mas muitos problemas como evitar a calcificação e estimular a condrogênese em meio favorável constituem o desafio para os pesquisadores na atualidade. 1) produzir anticorpo monoclonal específico a CTMs de coelho para monitorá-las, 2) determinar o volume ideal de coleta de medula óssea para microencapsulação e condrogênese, 3) realizar a microencapsulação das CTMs em novos arcabouços: BIOGEL3D e BACTCELL3D, comparando seu desempenho com o modelo clássico em alginato. Foram utilizados 25 coelhos Nova Zelândia sendo divididos em diferentes grupos em função do volume de MO coletado: G1 = 6mL, G2 = 9mL, G3 = 12mL, G4 = 15mL e G5 = volume ideal de coleta determinado pelos indicadores dos outros grupos. O material coletado foi diluído 1:2 em RPMI 1640 com 3.000U de heparina sódica. Após a contagem celular, as amostras foram submetidas a separação em gradiente... / Technological advances in the medical area combined with the impact of health programs that enhance longevity, together with lower natality rates created new challenges to the public manager such as aging and life as a “chronic disease”. Among the major problems are the chronic degenerative diseases that increase articular lesions. The limited regeneration capabilities and the limitations of actual treatment alternatives made biotechnology derived techniques the focus of investigations. The autologous chondrocyte transplantation (ACT) requires a small biopsy of health cartilage, which can lead to a new lesion. However, the use of stem cells can avoid this possibility. Mesenchymal stem cells (MSCs) can differentiate into chondrocytes by using a specific culture medium together with a 3D scaffold, but some questions such as the risks of calcification remain as key factors to researchers. 1) to produce a monoclonal antibody that recognizes MSCs in order to characterize them, 2) to determine the optimal bone marrow collection volume for cell microencapsulation and chondrogenesis and 3) to microencapsulate MSCs in two novel scaffolds: BIOGEL3D and BACTCELL3D, comparing them with sodium alginate. 25 New Zealand rabbits were divided into 5 groups related to bone marrow collection volume: G1 = 6mL, G2 = 9mL, G3 = 12mL, G4 = 15mL and G5 = optimal volume determined by the study. The collected material was diluted in RPMI1640 medium 1:2, with 3000U sodium heparin. After cell count and viability assessment the samples were submitted to density gradient centrifugation in order to isolate the lymphomononuclear (LMN) fraction. These cells were seeded to obtain and expand the MSCs in DMEM Knockout® (InvitrogenTM) supplemented with antibiotic/antimycotic, Lglutamine, essential aminoacids, non essential aminoacids and fetal bovine serum (all from InvitrogenTM). The cells were cultivated in 5% CO2 ...(Complete abstract click electronic access below)

Submitted by MARINA LANDIM E ALVARENGA null (marina@fmvz.unesp.br) on 2016-06-10T20:23:13Z No. of bitstreams: 1 dissertacao final.pdf: 2069659 bytes, checksum: f8308625d294fd21d8e9fadd7f344c09 (MD5) / Rejected by Ana Paula Grisoto (grisotoana@reitoria.unesp.br), reason: Solicitamos que realize uma nova submissão seguindo a orientação abaixo: O arquivo submetido está sem a ficha catalográfica. A versão submetida por você é considerada a versão final da dissertação/tese, portanto não poderá ocorrer qualquer alteração em seu conteúdo após a aprovação. Corrija esta informação e realize uma nova submissão contendo o arquivo correto. Agradecemos a compreensão. on 2016-06-15T18:05:22Z (GMT) / Submitted by MARINA LANDIM E ALVARENGA null (marina@fmvz.unesp.br) on 2016-06-26T01:29:11Z No. of bitstreams: 1 dissertacao final ficha.pdf: 2083568 bytes, checksum: 389a90011381796dbfd42b7a5be2f131 (MD5) / Approved for entry into archive by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br) on 2016-06-27T14:25:14Z (GMT) No. of bitstreams: 1 alvarenga_ml_me_bot.pdf: 2083568 bytes, checksum: 389a90011381796dbfd42b7a5be2f131 (MD5) / Made available in DSpace on 2016-06-27T14:25:14Z (GMT). No. of bitstreams: 1 alvarenga_ml_me_bot.pdf: 2083568 bytes, checksum: 389a90011381796dbfd42b7a5be2f131 (MD5) Previous issue date: 2016-04-26 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A utilização de terapias celulares no tratamento de lesões musculoesqueléticas cresce mundialmente em larga escala e é cada vez mais aceita. As células tronco mesenquimais (CTMs) são facilmente isoladas e expandidas in vitro, sendo tipicamente obtidas do próprio paciente (autóloga). Contudo, o uso de células provenientes de um animal doador (alogênica) e submetidas ao cultivo e a criopreservação, cria a oportunidade de iniciar o tratamento imediatamente após o diagnóstico. Há evidencias de que as CTMs possuem capacidade de imunomodulação, interagindo com os linfócitos T de diversas maneiras, além de não estimularem a indução de uma resposta imune complexa por não expressarem MHC II. Contudo, ainda há falta de consenso entre os estudos quanto a imunogenicidade das CTMs. Desta forma, o presente estudo objetivou avaliar a segurança da administração das células alogênicas em equinos através da análise de uma possível inflamação decorrente do implante alogênico de CTMs em músculo hígido de equinos, por meio de exame físico, ultrassonográfico, termográfico, histopatológico e pela expressão gênica de IL-1 beta e TNF- alfa. Os resultados demostraram que aplicação de CTM alogênicas é um processo seguro uma vez que não houveram alterações clínicas significativas, contudo uma segunda aplicação de CTM alogênicas de um mesmo doador levou ao aumento significativo da expressão gênica de TNF- alfa, podendo caracterizar uma resposta imune celular mais acentuada. Nossos resultados contribuíram para pesquisas futuras sobre tratamento de lesões musculoesqueléticas em equinos com CTMs, através da criação de bancos de células oriundas de doadores saudáveis. / The use of cell therapy for treatment of musculoskeletal injuries in horses is increasing in large scale worldwide. The mesenchymal stem cells (MSCs) are easily isolated and expanded in vitro, making them promising for use in clinical trials. Typically, these cells are obtained from the own patient (autologous), however, cell from a donor horse (allogeneic) expanded in vitro and cryopreserved create the opportunity of immediate treatment after diagnosis. There is evidence that MSCs have immunomodulatory capacity by interacting with T lymphocytes in several ways, and do not stimulate a complex immune response for not expressing MHC II. However, there is still a lack of consensus among the studies on the immunogenicity of MSCs. Thus, this study aimed to evaluate the safety of administration of allogeneic mesenchymal stem cells in horses. For such a possible inflammation reaction to the implantation was analysed through physical examination, ultrasound, thermography, histopathology and gene expression of IL 1beta and TNF-alpha. The results showed that the implantation of allogeneic MSCs is a safe process since there were no significant clinical changes in the animals, but a second application of allogeneic MSCs from the same donor led to significant increase in gene expression of TNF-alpha, which can characterize a cellular immune response. Our results contribute to future research on treatment of musculoskeletal injuries in horses using MSCs, by creating banks of cells derived from healthy donors.

Submitted by Ana Carolina Picolo Pasian (ana_781@hotmail.com) on 2018-04-20T19:54:18Z No. of bitstreams: 1 PARA DESPÓSITO.pdf: 1930882 bytes, checksum: 397a91ec261857e9a2bea286ae3eeb52 (MD5) / Approved for entry into archive by ROSANGELA APARECIDA LOBO null (rosangelalobo@btu.unesp.br) on 2018-04-23T13:24:13Z (GMT) No. of bitstreams: 1 pasian_acp_me_bot.pdf: 1930882 bytes, checksum: 397a91ec261857e9a2bea286ae3eeb52 (MD5) / Made available in DSpace on 2018-04-23T13:24:13Z (GMT). No. of bitstreams: 1 pasian_acp_me_bot.pdf: 1930882 bytes, checksum: 397a91ec261857e9a2bea286ae3eeb52 (MD5) Previous issue date: 2018-02-27 / A medicina regenerativa é uma área em crescente expansão no Brasil e no mundo, a qual procura ampliar a capacidade natural de regeneração dos tecidos através da utilização de células, fatores de proliferação e biomateriais. Um dos ramos da medicina regenerativa é a terapia celular, vertente que utiliza células-tronco, visando a substituição de tecidos funcionalmente ou estruturalmente lesados, apresentando um caráter terapêutico. Na medicina LASERs e LEDs vem sendo estudados como ferramenta terapêutica, mostrando possuir capacidade bioestimulatória. Este campo é caracterizado por uma variedade de metodologias, que são utilizadas em uma gama considerável de aplicações. Na técnica de fotoestimulação, utiliza-se a luz para ativar moléculas e funções celulares, apresentando o potencial de afetar a proliferação e diferenciação e o metabolismo da célula, estimulando a fosforilação oxidativa e podendo reduzir a resposta inflamatória local. Entretanto para que essa resposta ocorra, inúmeros trabalhos afirmam sobre a importância da seleção de um comprimento de onda ideal, uma vez que a utilização de um comprimento inapropriado pode acarretar em resultados contrários aos esperados, como a bioinibição. Diante destes achados o presente trabalho propôs-se a avaliar a ação do LED 850nm, no regime pulsátil, nas doses de 3, 5 e 10J/cm² na cultura de célula-tronco mesenquimal (CTM) com Soro Fetal Bovino (SFB) e com Hormônios derivados de plaquetas (HDP), e na cultura de células de Linfoma linfoblástico tipo B, RAJI Cells na dose de 10J/cm². Em ambos experimentos de exposição a luz, o comprimento de onda 850 nm inibiu a proliferação celular. Na cultura de CTM o LED tornou o desdobramento celular mais lento e acarretou na diminuição da confluência celular, especialmente nas doses de 5 e 10J/cm². Na cultura de linfoma Linfoblástico tipo B, em apenas 1 semana de exposição o mesmo comportamento de bioinibição foi encontrado na dose de 10J/cm². O grupo não tratado apresentou 7,0 X 10 5 células, em média, por frasco enquanto que as células submetidas à irradiação sofreram diminuição do tempo de desdobramento sendo a concentração destas de 4,2X105 , em média, por frasco. / Regenerative medicine is a promising growing area worldwide, with the aim of restore and regenerate tissues and whole organs through the use of cells, proliferation factors and biomaterials. One branch of regenerative medicine is cell therapy, that uses stem cells, aiming at the substitution of functionally or structurally damaged tissues, presenting therapeutic fature. LASERs and LEDs are available as therapeutic tools, showing biostimulating ability. The photo-stimulation technique uses light to activate molecules and cellular functions, presenting potential to affect proliferation, cell differentiation and metabolism, stimulating oxidative phosphorylation and reducing the local inflammatory response. Data shows the importance of selecting an ideal wavelength, such as the use of an inappropriate choice, can lead to undisered results, such as bioinhibition. In the present work, we evaluated the action of LED 850nm, pulsatile, at the doses of 3, 5 and 10J/cm² in mesenchymal stem cells (CTM) with Bovine Fetal Serum (FBS) and with derived platelets – Hormones (HDP) and B-cell lymphoblastic cell culture, 10J /cm2 , RAJI cells. In all light exposure experiments, wavelength of 850 nm inhibited cell proliferation. CTM culture, LED had a low proliferation rate, resulting in a decrease in cellular confluence, especially at 5 and 10J/cm2 . Lymphoblastic lymphoma type B cells, in only one week of exposure presente the same behavior of bioinhibition at 10J/cm2 . The control group had 7.0 x 105 cells on per vial, while cells subjected to irradiation underwent the unfolding time at a concentration of 4.2 x 105 on average per vial.