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1.
J Am Dent Assoc ; 154(12): 1048-1057, 2023 12.
Artigo em Inglês | MEDLINE | ID: mdl-37804275

RESUMO

BACKGROUND: Stem cells are present in most of the tissues in the craniofacial complex and play a major role in tissue homeostasis and repair. These cells are characterized by their capacity to differentiate into multiple cell types and to self-renew to maintain a stem cell pool throughout the life of the tissue. TYPES OF STUDIES REVIEWED: The authors discuss original data from experiments and comparative analyses and review articles describing the identification and characterization of stem cells of the oral cavity. RESULTS: Every oral tissue except enamel, dentin, and cementum contains stem cells for the entire life span. These stem cells self-renew to maintain a pool of cells that can be activated to replace terminally differentiated cells (for example, odontoblasts) or to enable wound healing (for example, dentin bridge in pulp exposures and healing of periodontal tissues after surgery). In addition, dental stem cells can differentiate into functional blood vessels and nerves. Initial clinical trials have shown that transplanting dental pulp stem cells into disinfected necrotic teeth has allowed for the recovery of tooth vitality and vertical and horizontal root growth in immature teeth with incomplete root formation. PRACTICAL IMPLICATIONS: As a consequence of these groundbreaking discoveries, stem cell banks are now offering services for the cryopreservation of dental stem cells. The future use of stem cell-based therapies in the clinic will depend on the collaboration of clinicians and researchers in projects designed to understand whether these treatments are safe, efficacious, and clinically feasible.


Assuntos
Polpa Dentária , Dente , Humanos , Polpa Dentária/metabolismo , Engenharia Tecidual , Células-Tronco/fisiologia , Odontologia
2.
J Dent Educ ; 87(3): 408-414, 2023 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-36200576

RESUMO

Dental education traditionally requires the use of dedicated fixed preclinic facilities to provide clinically relevant experiences to support the development of dexterity, critical thinking, and self-assessment skills that are essential for excellent patient care. As a result of the social distancing guidelines instituted at the height of the COVID pandemic, dental education was severely affected when education pivoted to remote instruction and had significant restrictions on in-person training. This study evaluated a novel application of modern technology to allow students to perform clinically relevant hands-on exercises away from dental school and, most importantly, receive feedback on their performance as an aid in their development. Student surveys and a comparison of pre- and post-COVID grades were used to evaluate the effectiveness of the virtual clinic to support remote dental education.


Assuntos
COVID-19 , Faculdades de Odontologia , Humanos , COVID-19/epidemiologia , Retroalimentação , Estudantes , Tecnologia
3.
Front Cell Dev Biol ; 10: 977725, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36340037

RESUMO

A functional vascular network requires that blood vessels are invested by mural cells. We have shown that dental pulp stem cells (DPSC) can undergo vasculogenic differentiation, and that the resulting vessels anastomize with the host vasculature and become functional (blood carrying) vessels. However, the mechanisms underlying the maturation of DPSC-derived blood vessels remains unclear. Here, we performed a series of studies to understand the process of mural cell investment of blood vessels generated upon vasculogenic differentiation of dental pulp stem cells. Primary human DPSC were co-cultured with primary human umbilical artery smooth muscle cells (HUASMC) in 3D gels in presence of vasculogenic differentiation medium. We observed DPSC capillary sprout formation and SMC recruitment, alignment and remodeling that resulted in complex vascular networks. While HUASMC enhanced the number of capillary sprouts and stabilized the capillary network when co-cultured with DPSC, HUASMC by themselves were unable to form capillary sprouts. In vivo, GFP transduced human DPSC seeded in biodegradable scaffolds and transplanted into immunodeficient mice generated functional human blood vessels invested with murine smooth muscle actin (SMA)-positive, GFP-negative cells. Inhibition of PDGFR-ß signaling prevented the SMC investment of DPSC-derived capillary sprouts in vitro and of DPSC-derived blood vessels in vivo. In contrast, inhibition of Tie-2 signaling did not have a significant effect on the SMC recruitment in DPSC-derived vascular structures. Collectively, these results demonstrate that PDGF-BB signaling via PDGFR-ß regulates the process of maturation (mural investment) of blood vessels generated upon vasculogenic differentiation of human dental pulp stem cells.

4.
Clin Cancer Res ; 28(21): 4757-4770, 2022 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-36048559

RESUMO

PURPOSE: Mucoepidermoid carcinoma (MEC) is a poorly understood salivary gland malignancy with limited therapeutic options. Cancer stem cells (CSC) are considered drivers of cancer progression by mediating tumor recurrence and metastasis. We have shown that clinically relevant small molecule inhibitors of MDM2-p53 interaction activate p53 signaling and reduce the fraction of CSC in MEC. Here we examined the functional role of p53 in the plasticity and self-renewal of MEC CSC. EXPERIMENTAL DESIGN: Using gene silencing and therapeutic activation of p53, we analyzed the cell-cycle profiles and apoptosis levels of CSCs in MEC cell lines (UM-HMC-1, -3A, -3B) via flow cytometry and looked at the effects on survival/self-renewal of the CSCs through sphere assays. We evaluated the effect of p53 on tumor development (N = 51) and disease recurrence (N = 17) using in vivo subcutaneous and orthotopic murine models of MEC. Recurrence was followed for 250 days after tumor resection. RESULTS: Although p53 activation does not induce MEC CSC apoptosis, it reduces stemness properties such as self-renewal by regulating Bmi-1 expression and driving CSC towards differentiation. In contrast, downregulation of p53 causes expansion of the CSC population while promoting tumor growth. Remarkably, therapeutic activation of p53 prevented CSC-mediated tumor recurrence in preclinical trials. CONCLUSIONS: Collectively, these results demonstrate that p53 defines the stemness of MEC and suggest that therapeutic activation of p53 might have clinical utility in patients with salivary gland MEC.


Assuntos
Carcinoma Mucoepidermoide , Neoplasias das Glândulas Salivares , Humanos , Camundongos , Animais , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo , Linhagem Celular Tumoral , Recidiva Local de Neoplasia/patologia , Neoplasias das Glândulas Salivares/patologia , Células-Tronco Neoplásicas/metabolismo , Carcinoma Mucoepidermoide/tratamento farmacológico , Carcinoma Mucoepidermoide/genética , Carcinoma Mucoepidermoide/metabolismo
5.
Eur J Dent Educ ; 26(2): 409-415, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-34520607

RESUMO

INTRODUCTION: Class IV composite restorations are one of the biggest challenges in dentistry. Furthermore, replacing adequate proximal contours on Class IV restorations is crucial for the function and aesthetics. The objective of this study is to assess four different teaching strategies used to improve first-year dental students' Class IV restoration proximal contact performance over a period of 4 years. MATERIALS AND METHODS: We assessed four cohorts of first-year dental students who were exposed to four different teaching strategies during the first-year preclinical training over two consecutive academic terms. The four different teaching strategies used were: (a) two waxing exercises (control cohort, strategy 1); (b) digital dentistry and four waxing exercises (strategy 2); (c) four waxing exercises (strategy 3); and (d) four waxing exercises and live demonstrations (strategy 4). All cohorts were exposed to the same didactic lecture of Class IV restorations. RESULTS: Our results showed that all teaching strategies resulted in better student's performance and content retention compared to the control cohort. However, the teaching strategy that resulted in the best pass/fail ratio was the association of waxing exercises with live demonstrations (strategy 4). DISCUSSION: Increasing the number of waxing exercises may improve students' performance either alone or associated with different teaching strategies. However, when associated with live demonstrations, waxing exercises have significantly reduced critical errors. CONCLUSIONS: Our study demonstrated for the first time the benefits of the affordable and traditional waxing exercises associated with instructor demonstrations as a teaching strategy for first-year dental students.


Assuntos
Educação em Odontologia , Estudantes de Odontologia , Estudos de Coortes , Educação em Odontologia/métodos , Avaliação Educacional/métodos , Humanos , Ensino
6.
Cells ; 10(11)2021 10 20.
Artigo em Inglês | MEDLINE | ID: mdl-34831027

RESUMO

Understanding how Mesenchymal Stem Cells (MSCs) form blood vessels is critical for creating mechanism-based approaches for the therapeutic use of these cells. In addition, understanding the determinants and factors involved in lineage hierarchy is fundamental to creating accurate and reliable techniques for the study of stem cells in tissue engineering and repair. Dental Pulp Stem Cells (DPSC) from permanent teeth and Stem cells from Human Exfoliated Deciduous teeth (SHED) are particularly interesting sources for tissue engineering as they are easily accessible and expandable. Previously, we have shown that DPSCs and SHEDs can differentiate into endothelial cells and form functional blood vessels through vasculogenesis. Here, we described how we created the "pulpbow" (pulp + rainbow), a multicolor tag experimental model that is stable, permanent, unique to each cell and passed through generations. We used the pulpbow to understand how dental pulp stem cells contributed to blood vessel formation in 3D models in in vitro and ex vivo live cell tracking, and in vivo transplantation assays. Simultaneous tracking of cells during sprout formation revealed that no single multicolor-tagged cell was more prone to vasculogenesis. During this process, there was intense cell motility with minimal proliferation in early time points. In later stages, when the availability of undifferentiated cells around the forming sprout decreased, there was local clonal proliferation mediated by proximity. These results unveiled that the vasculogenesis process mediated by dental pulp stem cells is dynamic and proximity to the sprouting area is critical for cell fate decisions.


Assuntos
Técnicas de Cultura de Células/métodos , Polpa Dentária/citologia , Células-Tronco Mesenquimais/citologia , Neovascularização Fisiológica , Animais , Cor , Masculino , Transplante de Células-Tronco Mesenquimais , Camundongos Endogâmicos C57BL , Camundongos SCID , Fatores de Tempo
8.
Cell Death Dis ; 12(1): 108, 2021 01 21.
Artigo em Inglês | MEDLINE | ID: mdl-33479203

RESUMO

Advanced salivary gland mucoepidermoid carcinoma (MEC) is a relentless cancer that exhibits resistance to conventional chemotherapy. As such, treatment for patients with advanced MEC is tipically radical surgery and radiotherapy. Facial disfigurement and poor quality of life are frequent treatment challenges, and many patients succumb to loco-regional recurrence and/or metastasis. We know that cancer stem-like cells (CSC) drive MEC tumorigenesis. The current study tests the hypothesis that MEC CSC are sensitive to therapeutic inhibition of mTOR. Here, we report a correlation between the long-term clinical outcomes of 17 MEC patients and the intratumoral expression of p-mTOR (p = 0.00294) and p-S6K1 (p = 0.00357). In vitro, we observed that MEC CSC exhibit constitutive activation of the mTOR signaling pathway (i.e., mTOR, AKT, and S6K1), unveiling a potential strategy for targeted ablation of these cells. Using a panel of inhibitors of the mTOR pathway, i.e., rapamycin and temsirolimus (mTOR inhibitors), buparlisib and LY294002 (AKT inhibitors), and PF4708671 (S6K1 inhibitor), we observed consistently dose-dependent decrease in the fraction of CSC, as well as inhibition of secondary sphere formation and self-renewal in three human MEC cell lines (UM-HMC-1,-3A,-3B). Notably, therapeutic inhibition of mTOR with rapamycin or temsirolimus induced preferential apoptosis of CSC, when compared to bulk tumor cells. In contrast, conventional chemotherapeutic drugs (cisplatin, paclitaxel) induced preferential apoptosis of bulk tumor cells and accumulation of CSC. In vivo, therapeutic inhibition of mTOR with temsirolimus caused ablation of CSC and downregulation of Bmi-1 expression (major inducer of stem cell self-renewal) in MEC xenografts. Transplantation of MEC cells genetically silenced for mTOR into immunodeficient mice corroborated the results obtained with temsirolimus. Collectively, these data demonstrated that mTOR signaling is required for CSC survival, and unveiled the therapeutic potential of targeting the mTOR pathway for elimination of highly tumorigenic cancer stem-like cells in salivary gland mucoepidermoid carcinoma.


Assuntos
Neoplasias de Cabeça e Pescoço/genética , Células-Tronco Neoplásicas/metabolismo , Neoplasias das Glândulas Salivares/genética , Serina-Treonina Quinases TOR/metabolismo , Neoplasias de Cabeça e Pescoço/patologia , Humanos , Neoplasias das Glândulas Salivares/patologia , Transdução de Sinais
9.
Braz Oral Res ; 34: e033, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32267290

RESUMO

The aim of our study was to isolate populations of keratinocyte stem cells based on the expression of cell surface markers and to investigate whether the culture could affect their phenotype. keratinocytes from human oral mucosa were sorted based on the expression of the epithelial stem cell markers p75NTR and CD71. We also examined the co-expression of other epithelial stem markers such as integrins ß1 and α6 and their stem cell-like proprieties in in vitro assays. Three passages after being sorted by MACS, more than 93% of the p75NTR+ve cells lost the expression of p75NTR, while 5.46% of the p75NTR-ve gained it. Within the small population of the p75NTR+ve cells, 88% co-expressed other epithelial stem cell markers such as integrins ß1 and α6, while only 28% of p75NTR-ve cells co-expressed these markers. These results were confirmed by sorting cells by FACS. Additionally, when double staining was used for sorting cells, 99% of the p75NTR+veCD71-ve and 33% of the p75NTR-veCD71+ve cells expressed both integrins, but just one week after culture, only 1.74% of the p75NTR+veCD71-ve cells still expressed p75NTR and only 0.32% still expressed CD71. Similar results were obtained when co-culturing p75NTR+ve and p75NTR-ve populations before analysis. Our results suggest that phenotype changes may be part of an intrinsic cellular mechanism to conserve levels of protein expression as they may found in the human body. In addition, in vitro culture may not offer ideal conditions for epithelial stem cell maintenance due to phenotype changes under standard culture conditions.


Assuntos
Técnicas de Cultura de Células/métodos , Células Epiteliais/citologia , Queratinócitos/citologia , Mucosa Bucal/citologia , Fenótipo , Células-Tronco/citologia , Antígenos CD/análise , Biomarcadores/análise , Separação Celular/métodos , Citometria de Fluxo/métodos , Humanos , Proteínas do Tecido Nervoso/análise , Receptores de Fator de Crescimento Neural/análise , Receptores da Transferrina/análise , Reprodutibilidade dos Testes
10.
Cell Tissue Bank ; 21(1): 31-46, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31807957

RESUMO

Dental stem cells have many applications in medicine, dentistry and stem cell biology in general due to their easy accessibility and low morbidity. A common surgical manoeuvre after a tooth extraction is the dental socket curettage which is necessary to clean the alveolus and favour alveolar bone healing. This procedure can cause very low morbidity compared to bone marrow collection procedures and the collected material is normally discarded. In order to investigate if the tissue obtained by dental socket curettage after a tooth extraction was a feasible alternative source to isolate human stem cells, we isolated and characterized two different stem cell populations based on STRO-1 and CD146 expression. We were able to collect and grow cells from dental socket of vital and non-vital teeth. Both populations were proliferative, clonogenic and expressed STRO-1, CD146, CD90, NG2, PDGFR-ß, which are markers found in stem cells, presented in vitro multiline-differentiation into osteogenic, chondrogenic, and adipogenic tissue, and in vivo transplanted cells formed mineralized tissue. Interestingly, STRO-1+ clonogenic cells presented better multidifferentiation than CD146+ cells. Our results showed that mesenchymal stem cells can be isolated from the tiny tissue collected by dental socket curettage after vital and non-vital tooth extraction and suggest that STRO-1 is an important marker to be used to sort cells with multidifferentiation capacity.


Assuntos
Diferenciação Celular , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/citologia , Alvéolo Dental/citologia , Animais , Antígenos de Superfície/análise , Antígeno CD146/análise , Proliferação de Células , Células Cultivadas , Humanos , Separação Imunomagnética , Masculino , Camundongos Endogâmicos BALB C , Camundongos Nus
11.
Braz. oral res. (Online) ; 34: e033, 2020. graf
Artigo em Inglês | LILACS | ID: biblio-1089391

RESUMO

Abstract The aim of our study was to isolate populations of keratinocyte stem cells based on the expression of cell surface markers and to investigate whether the culture could affect their phenotype. keratinocytes from human oral mucosa were sorted based on the expression of the epithelial stem cell markers p75NTR and CD71. We also examined the co-expression of other epithelial stem markers such as integrins β1 and α6 and their stem cell-like proprieties in in vitro assays. Three passages after being sorted by MACS, more than 93% of the p75NTR+ve cells lost the expression of p75NTR, while 5.46% of the p75NTR-ve gained it. Within the small population of the p75NTR+ve cells, 88% co-expressed other epithelial stem cell markers such as integrins β1 and α6, while only 28% of p75NTR-ve cells co-expressed these markers. These results were confirmed by sorting cells by FACS. Additionally, when double staining was used for sorting cells, 99% of the p75NTR+veCD71-ve and 33% of the p75NTR-veCD71+ve cells expressed both integrins, but just one week after culture, only 1.74% of the p75NTR+veCD71-ve cells still expressed p75NTR and only 0.32% still expressed CD71. Similar results were obtained when co-culturing p75NTR+ve and p75NTR-ve populations before analysis. Our results suggest that phenotype changes may be part of an intrinsic cellular mechanism to conserve levels of protein expression as they may found in the human body. In addition, in vitro culture may not offer ideal conditions for epithelial stem cell maintenance due to phenotype changes under standard culture conditions.


Assuntos
Humanos , Fenótipo , Células-Tronco/citologia , Queratinócitos/citologia , Técnicas de Cultura de Células/métodos , Células Epiteliais/citologia , Mucosa Bucal/citologia , Receptores da Transferrina/análise , Biomarcadores/análise , Antígenos CD/análise , Separação Celular/métodos , Reprodutibilidade dos Testes , Receptores de Fator de Crescimento Neural/análise , Citometria de Fluxo/métodos , Proteínas do Tecido Nervoso/análise
12.
J Endod ; 44(11): 1671-1676, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30409448

RESUMO

INTRODUCTION: Dental pulp is a complex tissue with highly differentiated cells, which makes its reconstruction a challenging task. The apical papilla is an undifferentiated tissue considered as the remnant of the dental papilla that forms the dentin-pulp complex. Aiming to analyze morphologic features of the tissue formed in an in vivo pulp model, we used human apical papilla as a cell source without the use of exogenous growth factors. METHODS: A construct was built using newborn mice molar crowns treated with TrypLE (Fisher Scientific, Loughborough, UK) and EDTA. The crowns were filled with PuraMatrix (Corning Inc, Corning, NY) and a pool population of human apical papilla cells. As a control, we used crowns filled only with PuraMatrix and empty crowns. The constructs were transplanted under severe combined immunodeficient mice kidney capsules. Immunohistochemistry for lamin A, dentin sialophosphoprotein, and dentin matrix protein 1 was performed. RESULTS: Morphologic analysis of all transplanted crowns showed the formation of a loose connective tissue of variable cellularity with the presence of well-formed functional vessels. In the study group, lamin A-positive cells represented the majority of cells within the pulp chamber and a few cells in the vessel lining. We also found positivity for dentin sialophosphoprotein and dentin matrix protein 1, an indicator of odontoblast differentiation. CONCLUSIONS: In our study model, human transplanted apical papilla cells mixed with the host cells and formed a vascularized viable tissue, and these cells were able to differentiate into odontoblastlike cells without the use of exogenous growth factors.


Assuntos
Diferenciação Celular , Papila Dentária/citologia , Papila Dentária/fisiologia , Polpa Dentária , Odontoblastos , Ápice Dentário/citologia , Ápice Dentário/fisiologia , Animais , Animais Recém-Nascidos , Diferenciação Celular/genética , Transplante de Células , Papila Dentária/transplante , Polpa Dentária/citologia , Polpa Dentária/fisiologia , Proteínas da Matriz Extracelular/metabolismo , Humanos , Peptídeos e Proteínas de Sinalização Intercelular , Camundongos Endogâmicos , Fosfoproteínas/metabolismo , Regeneração , Sialoglicoproteínas/metabolismo , Ápice Dentário/transplante
13.
Cell Tissue Bank ; 18(2): 217-228, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28233169

RESUMO

Several non-biological materials are currently being used to increase the alveolar bone volume to support dental implants. Recently, stem cell therapy has emerged as a promising biological substitute or adjuvant to enhance bone healing. In order to determine if stem cell therapy has enough clinical evidence to bone ridge augmentation in humans, a systematic review and meta-analysis were conducted. Two independent investigators searched the Entrez PubMed, SCOPUS and Web of Science databases for eligible randomized clinical trials that describe stem cell therapies for alveolar bone formation. The included studies were evaluated for risk of bias. A random-effects meta-analysis model was used to evaluate the percentage of bone formation in the selected studies. Heterogeneity was evaluated using the Cochrane Chi 2 and I 2. Nine eligible trials were included. These studies presented an overall unclear risk of bias. A comparison between the lower heterogeneity studies and the long term observational outcomes showed a slight tendency to enhance bone formation. High heterogeneity between the included studies was observed. The lack of outcome standardization made a wide-ranging comparison difficult. The application of stem cells in oral surgery and implantology appears to be promising although more standardized study designs, increased samples and long-term observations are needed to strength the clinical evidence that stem cell therapy is effective for alveolar bone formation.


Assuntos
Processo Alveolar/fisiologia , Processo Alveolar/cirurgia , Aumento do Rebordo Alveolar/métodos , Implantação Dentária Endóssea/métodos , Osteogênese , Transplante de Células-Tronco/métodos , Células-Tronco/citologia , Processo Alveolar/citologia , Implantes Dentários , Humanos , Ensaios Clínicos Controlados Aleatórios como Assunto , Resultado do Tratamento
14.
Braz Oral Res ; 30(1): e109, 2016 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-27737362

RESUMO

Multicystic and unicystic ameloblastomas are benign odontogenic tumors that present distinct biological behavior. The investigation of stem cells has become an important branch of tumor biology, with several studies addressing the possible role of these cells in tumor growth, angiogenesis, progression, infiltration and invasiveness. This study evaluated the immunohistochemical expression of CD90(Thy-1) and P75NTR stem cell markers in multicystic and unicystic ameloblastomas. Seventeen (17) samples of ameloblastomas (multicystic, n = 10; unicystic, n = 7) were submitted to immunohistochemical reactions and graded semi-quantitatively. The Kolmogorov-Smirnov test was used to verify possible differences in CD90 and P75NTR expressions between multicystic and unicystic ameloblastomas (p < 0.05). CD90 immunostaining was observed in all multicystic ameloblastoma specimens (n = 10), in the cytoplasm of the fibroblasts and vascular endothelial cells of the tumor stroma, near the neoplastic odontogenic epithelia. The staining of stromal CD90 was significantly higher in multicystic than in unicystic ameloblastomas (p = 0.003). Nuclear P75NTR immunostaining was observed in all ameloblastoma specimens. A significant difference was seen in the epithelial staining of P75NTR between multicystic and unicystic types (p = 0.007). The increased expression of CD90 and P75NTR found in multicystic ameloblastomas suggests a behavioral biological difference between multicystic and unicystic ameloblastomas, as well as a difference in ameloblastoma development.


Assuntos
Ameloblastoma/metabolismo , Biomarcadores Tumorais/metabolismo , Neoplasias Mandibulares/metabolismo , Células-Tronco Neoplásicas/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Receptores de Fator de Crescimento Neural/metabolismo , Antígenos Thy-1/metabolismo , Adolescente , Adulto , Ameloblastoma/patologia , Células Endoteliais/metabolismo , Feminino , Fibroblastos/metabolismo , Humanos , Imuno-Histoquímica , Masculino , Neoplasias Mandibulares/patologia , Pessoa de Meia-Idade , Células-Tronco Neoplásicas/patologia , Inclusão em Parafina , Estatísticas não Paramétricas , Células Estromais/metabolismo , Adulto Jovem
15.
Microsc Res Tech ; 79(3): 149-54, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26749077

RESUMO

The micro-X-ray fluorescence by synchrotron radiation (µ-XRF) is a method to determine the composition of tissues without destroying the samples. However, this technique has never been used for the analysis of mesenchymal stem cells (MSC). This study compared different protocols for fixing, storing, preserving, and establishing the correct numbers of dental derived MSC submitted to µ-XRF analysis. Stem cells were obtained from human dental tissue. After cell expansion, and MACS isolation, the samples were fixed and the following quantities of cells 1 × 10(4) to 1 × 10(7) were divided in two groups: G1: fixed in 4% paraformaldehyde diluted in phosphate-buffered saline solution, and G2: fixed in 4% paraformaldehyde diluted in MilliQ water. The G1 cells showed precipitation of chemical components from the solution resulting in the formation of salt crystals while G2 cells were clear and almost transparent in the sample holder. With regards to cells concentration, the best results occurred when four droplets of 1 × 10(7) cells were analyzed. This work shows that to identify and study the distribution of trace elements in MSC by µ-XRF, the best protocol is fixation in 4% paraformaldehyde diluted with MilliQ water at 4°C and a concentration of four incremental droplets of 1 × 10(7) cells.


Assuntos
Histocitoquímica/métodos , Células-Tronco Mesenquimais/química , Células-Tronco Mesenquimais/citologia , Microscopia de Fluorescência/métodos , Espectrometria por Raios X/métodos , Células Cultivadas , Humanos , Metais/análise , Metais/química , Síncrotrons
16.
Braz. oral res. (Online) ; 30(1): e109, 2016. tab, graf
Artigo em Inglês | LILACS | ID: biblio-952054

RESUMO

Abstract Multicystic and unicystic ameloblastomas are benign odontogenic tumors that present distinct biological behavior. The investigation of stem cells has become an important branch of tumor biology, with several studies addressing the possible role of these cells in tumor growth, angiogenesis, progression, infiltration and invasiveness. This study evaluated the immunohistochemical expression of CD90(Thy-1) and P75NTR stem cell markers in multicystic and unicystic ameloblastomas. Seventeen (17) samples of ameloblastomas (multicystic, n = 10; unicystic, n = 7) were submitted to immunohistochemical reactions and graded semi-quantitatively. The Kolmogorov-Smirnov test was used to verify possible differences in CD90 and P75NTR expressions between multicystic and unicystic ameloblastomas (p < 0.05). CD90 immunostaining was observed in all multicystic ameloblastoma specimens (n = 10), in the cytoplasm of the fibroblasts and vascular endothelial cells of the tumor stroma, near the neoplastic odontogenic epithelia. The staining of stromal CD90 was significantly higher in multicystic than in unicystic ameloblastomas (p = 0.003). Nuclear P75NTR immunostaining was observed in all ameloblastoma specimens. A significant difference was seen in the epithelial staining of P75NTR between multicystic and unicystic types (p = 0.007). The increased expression of CD90 and P75NTR found in multicystic ameloblastomas suggests a behavioral biological difference between multicystic and unicystic ameloblastomas, as well as a difference in ameloblastoma development.


Assuntos
Humanos , Masculino , Feminino , Adolescente , Adulto , Adulto Jovem , Células-Tronco Neoplásicas/metabolismo , Ameloblastoma/metabolismo , Neoplasias Mandibulares/metabolismo , Biomarcadores Tumorais/metabolismo , Receptores de Fator de Crescimento Neural/metabolismo , Antígenos Thy-1/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Células-Tronco Neoplásicas/patologia , Imuno-Histoquímica , Ameloblastoma/patologia , Neoplasias Mandibulares/patologia , Inclusão em Parafina , Células Estromais , Estatísticas não Paramétricas , Células Endoteliais/metabolismo , Fibroblastos/metabolismo , Pessoa de Meia-Idade
17.
Oncotarget ; 6(29): 26633-50, 2015 Sep 29.
Artigo em Inglês | MEDLINE | ID: mdl-26449187

RESUMO

A small sub-population of cells characterized by increased tumorigenic potential, ability to self-renew and to differentiate into cells that make up the tumor bulk, has been characterized in some (but not all) tumor types. These unique cells, namedcancer stem cells, are considered drivers of tumor progression in these tumors. The purpose of this work is to understand if cancer stem cells play a functional role in the tumorigenesis of salivary gland mucoepidermoid carcinomas. Here, we investigated the expression of putative cancer stem cell markers (ALDH, CD10, CD24, CD44) in primary human mucoepidermoid carcinomas by immunofluorescence, in vitro salisphere assays, and in vivo tumorigenicity assays in immunodeficient mice. Human mucoepidermoid carcinoma cells (UM-HMC-1, UM-HMC-3A, UM-HMC-3B) sorted for high levels of ALDH activity and CD44 expression (ALDHhighCD44high) consistently formed primary and secondary salispheres in vitro, and showed enhanced tumorigenic potential in vivo (defined as time to tumor palpability, tumor growth after palpability), when compared to ALDHlowCD44low cells. Cells sorted for CD10/CD24, and CD10/CD44 showed varying trends of salisphere formation, but consistently low in vivo tumorigenic potential. And finally, cells sorted for CD44/CD24 showed inconsistent results in salisphere formation and tumorigenic potential assays when different cell lines were evaluated. Collectively, these data demonstrate that salivary gland mucoepidermoid carcinomas contain a small population of cancer stem cells with enhanced tumorigenic potential and that are characterized by high ALDH activity and CD44 expression. These results suggest that patients with mucoepidermoid carcinoma might benefit from therapies that ablate these highly tumorigenic cells.


Assuntos
Carcinoma Mucoepidermoide/metabolismo , Regulação Neoplásica da Expressão Gênica , Receptores de Hialuronatos/metabolismo , Isoenzimas/metabolismo , Células-Tronco Neoplásicas/metabolismo , Retinal Desidrogenase/metabolismo , Neoplasias das Glândulas Salivares/metabolismo , Adolescente , Adulto , Idoso , Família Aldeído Desidrogenase 1 , Animais , Antígeno CD24/metabolismo , Linhagem Celular Tumoral , Feminino , Citometria de Fluxo , Humanos , Masculino , Camundongos , Camundongos SCID , Microcirculação , Microscopia de Fluorescência , Pessoa de Meia-Idade , Transplante de Neoplasias , Neprilisina/metabolismo
18.
BMC Res Notes ; 7: 673, 2014 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-25252856

RESUMO

BACKGROUND: Gingival hyperplasias are peculiar conditions that may produce extreme growth that impairs masticatory function and causes psychological and aesthetic disturbances. They can vary from mild interdental papillae localized growth to marked swelling affecting both jaws. CASE PRESENTATION: The aim of this case report is to present a rare case of generalized gingival growth diagnosed in a 4 year-old Caucasian child and followed for 9 years. The lesion covered almost all of the upper and lower teeth and recurred thirty times with the same clinical and histopathological aspects. The clinical features suggested the diagnosis of idiopathic gingival fibromatosis, but the histopathological aspects did not confirm this hypothesis and were consistent with peripheral ossifying fibroma. CONCLUSION: The present case reports a rare gingival growth with challenging diagnosis and treatment.


Assuntos
Fibroma/diagnóstico , Neoplasias Gengivais/diagnóstico , Pré-Escolar , Diagnóstico Diferencial , Fibroma/diagnóstico por imagem , Fibroma/patologia , Neoplasias Gengivais/diagnóstico por imagem , Neoplasias Gengivais/patologia , Humanos , Masculino , Radiografia , Recidiva
19.
Rev. odontol. UNESP (Online) ; 43(5): 326-332, Sep-Oct/2014. tab, ilus
Artigo em Inglês | LILACS, BBO - Odontologia | ID: lil-721487

RESUMO

Introduction: Dental fluorosis is a disturbance of high prevalence caused by the ingestion of fluoride ions present mainly in toothpaste. Preventive measures to avoid it are still controversial. Thus, knowing the impact that fluorosis can cause on the population's quality of life it is important for planning public health policies. Objective: To evaluate the impact of dental fluorosis on the quality of life of children and adolescents. Material and Method: We studied 300 subjects aged 8 to 12 years divided into 4 groups: children (8-10 years) and adolescents (10 to 12 years) with and without fluorosis. The diagnosis of fluorosis was performed according to the index Thylstrup and Fejerskov and quality of life was evaluated using Child Perceptions Questionnaire 8-10 and 11-14. The socio-demographic characteristics of the patients were also evaluated. For inclusion in the sample, selected patients should present eight permanent incisors with crowns fully erupted. Patients who had extensive restorations, fractured teeth, other dental enamel defects and who wore braces were excluded. Result: Fluorosis was present in 64.7% of the patients analyzed and in most cases (80.3%) was mild or very mild. In children, the average overall score of the questionnaire was 15.9 for the group without fluorosis and 18.3 for the group with fluorosis (p = 0.255). The teenagers' score in the group without fluorosis was 26.1, while the group with fluorosis was 22.7 (p = 0.104). Conclusion: Dental fluorosis caused impact on the quality of life of the population analyzed only in the functional domain. .


Introdução: A fluorose dentária é um distúrbio de alta prevalência decorrente da ingestão de íons fluoretos. Medidas preventivas para evitá-la ainda são controversas. Assim, conhecer o impacto que a fluorose pode causar na qualidade de vida de indivíduos é importante para o planejamento de políticas públicas de saúde. Objetivo: Avaliar o impacto da fluorose dentária sobre a qualidade de vida relacionada à saúde bucal (QVRSB) de crianças e adolescentes. Material e Método: Foram avaliados 300 indivíduos na faixa etária de 8 a12 anos. O diagnóstico de fluorose foi realizado segundo o índice Thylstrup e Fejerskov e a qualidade de vida foi avaliada utilizando os questionários de Percepção da Criança 8-10 e 11-14. Foram incluídos pacientes com oito incisivos permanentes com coroas totalmente irrompidase excluídos os que apresentavam restaurações extensas, dentes fraturados, outros defeitos do esmalte dentário e os que usavam aparelho ortodôntico fixo. Os dados foram analisados no programa SPSS(r) (versão 18; Chicago, IL) e realizaram-se os teste Qui-quadrado, Fisher e Mann-Whitney. Foram considerados significantes valores de p<0,05. Resultado: A prevalência de fluorose foi 64,7%, sendo os graus leve e muito leve responsáveis por 80,3% dos casos. Crianças e adolescentes não tiveram impacto na QVRSB no escore geral e domínios sintomas orais, bem-estar emocional e social (p>0,05). Entretanto, apresentaram impacto no domínio limitação funcional (p = 0,039 e 0,013) para crianças e adolescentes respectivamente). Conclusão: Foi observada associação entre fluorose e qualidade de vida apenas no domínio funcional. .


Assuntos
Humanos , Masculino , Feminino , Criança , Percepção , Qualidade de Vida , Distribuição de Qui-Quadrado , Estatísticas não Paramétricas , Política de Saúde , Fluorose Dentária
20.
Braz Oral Res ; 282014.
Artigo em Inglês | MEDLINE | ID: mdl-25166769

RESUMO

Dental pulp has been identified as a novel and promising stem cell source. The following systematic review presents and summarises in vivo studies that have used stem cells from the dental pulp of permanent and deciduous teeth to repair or regenerate non-dental tissues. An electronic customised search was performed using 4 different databases (Entrez PubMed, Cab Abstracts, Scopus and Web of Science). Only full-text research manuscripts published in English between the years of 2000 and 2012 were included. The manuscripts were retrieved based on the following keywords and/or abbreviations: [Stem Cells from Human Exfoliated Deciduous teeth (SHED)] AND/OR [Dental Pulp Stem Cells (DPSC)] AND [tissue regeneration] AND [tissue repair]. Only manuscripts involving in vivo applications of SHED or DPSC for the repair and/or regeneration of non-dental tissues were included. The search strategy produced 2309 papers, from which 14 were eligible according to the predetermined inclusion and exclusion criteria. Although human tissue was the source of cells in half of the studies included in our review, all of the studies involved transplantation into animals of other species, such as pigs, rats and mice. Most of the manuscripts reported the successful use of DPSCs or SHED for non-dental tissue repair or regeneration. While these cell populations represent promising alternative sources of stem cells for tissue engineering and cell-based regenerative medicine therapies, it is not yet possible to guarantee the appropriate clinical management of this technique.


Assuntos
Polpa Dentária/citologia , Regeneração/fisiologia , Transplante de Células-Tronco/métodos , Células-Tronco/citologia , Dente Decíduo/citologia , Animais , Regeneração Óssea/fisiologia , Cães , Humanos , Camundongos , Coelhos , Ratos , Reprodutibilidade dos Testes , Suínos , Engenharia Tecidual/métodos
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