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Minimally invasive exodontia is among the long-sought-for development aims of safe dental medicine. In this paper, we aim, for the first time, to examine whether the enzymatic disruption of the periodontal ligament fibers reduces the force required for tooth extraction. To this end, recombinantly expressed clostridial collagenase G variant purified from Escherichia coli was injected into the periodontal ligament of mesial and distal roots of the first and second split porcine mandibular premolars. The vehicle solution was injected into the corresponding roots on the contralateral side. Following sixteen hours, the treated mandibles were mounted on a loading machine to measure the extraction force. In addition, the effect of the enzyme on the viability of different cell types was evaluated. An average reduction of 20% in the applied force (albeit with a large variability of 50 to 370 newton) was observed for the enzymatically treated roots, reaching up to 50% reduction in some cases. Importantly, the enzyme showed only a minor and transient effect on cellular viability, without any signs of toxicity. Using an innovative model enabling the analytical measurement of extraction forces, we show, for the first time, that the enzymatic disruption of periodontal ligament fibers substantially reduces the force required for tooth extraction. This novel technique brings us closer to atraumatic exodontia, potentially reducing intra- and post-operative complications and facilitating subsequent implant placement. The development of novel enzymes with enhanced activity may further simplify the tooth extraction process and present additional clinical relevance for the broad range of implications in the oral cavity.
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INTRODUCTION: Identification of the dental arch form of the orthodontic patient is a key aspect for achieving a stable, functional, and esthetic dentition. Failure to customize preformed archwires with the patient's arch form might increase the probability of relapse and lead to an unnatural smile. The primary objective of this study was to identify the arch forms of Israeli subjects with dental normocclusion and malocclusions. The secondary objective was to clarify the morphologic differences between Israeli and North American white subjects with various malocclusions. METHODS: The sample included 134 Israeli (40 Class I, 61 Class II, and 33 Class III) and 160 North American (60 Class I, 50 Class II, and 50 Class III) subjects. The most facial portion of 13 proximal contact areas was digitized from photocopied images of the mandibular dental arches. Clinical bracket points were calculated for each tooth based on mandibular tooth thickness data. Four linear and 2 proportional measurements were made. The dental arches were classified as square, ovoid, and tapered forms to determine and compare the frequency distributions between the 2 ethnic groups. RESULTS: The most frequent mandibular arch form of the Israeli group was found to be ovoid as opposed to tapered in the North American white group. The white population had statistically significant decreased arch widths and increased arch depths compared with the Israeli population. The analysis of the Israeli sample indicated that, as the malocclusion shifts from Class III through Class I to Class II, the weight of the arch form tends to shift from square and ovoid to ovoid and tapered. CONCLUSIONS: Our results suggest that, when treating Israeli patients, one should expect to use the preformed ovoid arch form orthodontic wires in a significant percentage of patients.
Assuntos
Arco Dental/anatomia & histologia , Etnicidade , Má Oclusão/patologia , Mandíbula/anatomia & histologia , Adolescente , Adulto , Dente Pré-Molar/anatomia & histologia , Cefalometria/métodos , Criança , Dente Canino/anatomia & histologia , Arco Dental/patologia , Feminino , Humanos , Processamento de Imagem Assistida por Computador , Incisivo/anatomia & histologia , Israel , Masculino , Má Oclusão Classe I de Angle/patologia , Má Oclusão Classe II de Angle/patologia , Má Oclusão Classe III de Angle/patologia , Mandíbula/patologia , Dente Molar/anatomia & histologia , América do Norte , Fotografia Dentária , População Branca , Adulto JovemRESUMO
The highly regenerative capacity of the human adult oral mucosa suggests the existence of a robust stem cell (SC) population in its lamina propria (OMLP). The purpose of this study was to characterize the availability, growth, immunophenotype, and potency of this presumable SC population. Cells positive for the embryonic stem cell transcription factors Oct4 and Sox2 and for p75 formed distinct cord-like structure in the OMLP. Regardless of donor age, trillions of cells, termed human oral mucosa stem cells (hOMSC), 95% of which express mesenchymal stromal cell markers, were simply, and reproducibly produced from a biopsy of 3-4 x 2 x 1 mm(3). A total of 40-60% of these cells was positive for Oct4, Sox2, and Nanog and 60-80% expressed constitutively neural and neural crest SC markers. hOMSC differentiated in culture into mesodermal (osteoblastic, chondroblastic, and adipocytic), definitive endoderm and ectodermal (neuronal) lineages. Unexpectedly, hOMSC treated with dexamethasone formed tumors consisting of two germ layer-derived tissues when transplanted in severe combined immune deficiency mice. The tumors consisted of tissues produced by neural crest cells during embryogenesis-cartilage, bone, fat, striated muscle, and neural tissue. These results show that the adult OMLP harbors a primitive SC population with a distinct primitive neural-crest like phenotype and identifies the in vivo localization of putative ancestors for this population. This is the first report on ectodermal- and mesodermal-derived mixed tumors formation by a SC population derived from a nonmalignant somatic adult human tissue.
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Células-Tronco Adultas/citologia , Células-Tronco Mesenquimais/citologia , Mucosa Bucal/citologia , Adulto , Células-Tronco Adultas/classificação , Células-Tronco Adultas/metabolismo , Idoso , Idoso de 80 Anos ou mais , Animais , Biomarcadores/análise , Biomarcadores/metabolismo , Diferenciação Celular/fisiologia , Linhagem da Célula/fisiologia , Transformação Celular Neoplásica/metabolismo , Células Cultivadas , Humanos , Células-Tronco Mesenquimais/classificação , Células-Tronco Mesenquimais/metabolismo , Camundongos , Camundongos SCID , Pessoa de Meia-Idade , Mucosa Bucal/fisiologia , Mucosa/citologia , Mucosa/fisiologia , Adulto JovemRESUMO
Monitoring gene expression in vitro and in vivo, is crucial when analyzing osteogenesis and developing effective bone gene therapy protocols. Until recently, molecular analytical tools were only able to detect protein expression either in vitro or in vivo. These systems include histology and immunohistochemistry, fluorescent imaging, PET (micro-PET), CT (micro-CT), and bioluminescent imaging. The last is the only system to date that can enable efficient quantitative monitoring of gene expression both in vitro and in vivo. Effective bioluminescent imaging in bone can be achieved by using transgenic mice harboring the luciferase reporter gene, downstream of an osteogenesis specific promoter. The aim of this chapter is to comprehensively describe the various protocols needed for the detection of bioluminescence in bone development and repair.
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Osso e Ossos/citologia , Proteínas Luminescentes/metabolismo , Microscopia de Fluorescência , Animais , Osso e Ossos/metabolismo , Humanos , Imuno-Histoquímica/métodos , Proteínas Luminescentes/genética , Camundongos , Camundongos Transgênicos , Microscopia de Fluorescência/instrumentação , Microscopia de Fluorescência/métodos , Osteocalcina/genética , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismoRESUMO
Fluorescence molecular tomography (FMT) is a novel tomographic near-infrared (NIR) imaging modality that enables 3D quantitative determination of fluorochrome distribution in tissues of live small animals at any depth. This study demonstrates a noninvasive, quantitative method of monitoring engineered bone remodeling via FMT. Murine mesenchymal stem cells overexpressing the osteogenic gene BMP2 (mMSCs-BMP2) were implanted into the thigh muscle and into a radial nonunion bone defect model in C3H/HeN mice. Real-time imaging of bone formation was performed following systemic administration of the fluorescent bisphosphonate imaging agent OsteoSense, an hydroxyapatite-directed bone-imaging probe. The mice underwent imaging on days 7, 14, and 21 postimplantation. New bone formation at the implantation sites was quantified using micro-computed tomography (micro-CT) imaging. A higher fluorescent signal occurred at the site of the mMSC-BMP2 implants than that found in controls. Micro-CT imaging revealed a mass of mature bone formed in the implantation sites on day 21, a finding also confirmed by histology. These findings highlight the effectiveness of FMT as a functional platform for molecular imaging in the field of bone regeneration and tissue engineering.
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Consolidação da Fratura/fisiologia , Fraturas não Consolidadas/terapia , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/fisiologia , Osteogênese/fisiologia , Fraturas do Rádio/terapia , Animais , Feminino , Corantes Fluorescentes , Fraturas não Consolidadas/diagnóstico por imagem , Fraturas não Consolidadas/patologia , Engenharia Genética , Camundongos , Camundongos Endogâmicos C3H , Fraturas do Rádio/diagnóstico por imagem , Fraturas do Rádio/patologia , Tomografia Computadorizada por Raios XRESUMO
Angiogenesis is mandatory for reperfusion of viable tissues, and lack of vascularization may cause ischemia. The increasing disparity between the demand and availability of adequate substitutes for small-diameter human blood vessels has prompted an intensive search for artificial materials or biological allograft tissues, both of which usually fail in the long term. The objective of this study was to pioneer a novel model for in vivo guided angiogenesis based on a specific design process of a filamentous polymeric scaffold with endothelial cells in a 3-dimensional culture system. To our knowledge, this is the first report of an in vivo guided angiogenesis approach based on a 2-step model, composed of endothelial cells and a filamentous polymeric scaffold framework. Endothelial cells that had been cultured on a specifically designed filamentous polymeric scaffold within a regulated dynamic tissue culture system were shown in vivo to induce guided angiogenesis. Cells seeded on a biodegradable polymeric scaffold were implanted into mice. On day 28 after implantation, analysis revealed a guided angiogenic process along the path of the implanted polymeric scaffold as well as initial evidence for early maturation of engineered vessels, allowing red blood cells to flow through the forming lumina of new vessels as the polymer degraded. The authors conclude that in vivo guided angiogenesis can be achieved by combining endothelial cells with biodegradable filamentous polymeric scaffolds and that this model can lay the cornerstone for vascular engineering and future development of clinically available protocols aimed to treat life-threatening cardiovascular conditions.
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Células Endoteliais/citologia , Neovascularização Fisiológica , Polímeros/química , Engenharia Tecidual/métodos , Actinas/metabolismo , Animais , Anticorpos Monoclonais/metabolismo , Reatores Biológicos , Neoplasias Encefálicas/patologia , Varredura Diferencial de Calorimetria , Caproatos/química , Células Cultivadas , Células Endoteliais/química , Células Endoteliais/transplante , Genes Reporter , Proteínas de Fluorescência Verde/metabolismo , Imuno-Histoquímica , Luciferases/metabolismo , Medições Luminescentes , Imageamento por Ressonância Magnética , Teste de Materiais , Camundongos , Camundongos Nus , Modelos Biológicos , Peso Molecular , Músculo Liso/metabolismo , Técnicas de Cultura de Órgãos , Polietilenoglicóis/química , Especificidade por Substrato , Fatores de Tempo , EstanhoRESUMO
Viral delivery of the therapeutic gene bone morphogenetic protein-2 (BMP-2) is a promising approach for bone regeneration. The human parvovirus adeno-associated virus (AAV) type 2 is considered one of the most encouraging viral vector systems because of its high transduction rates and biosafety ratings. Bone morphogenetic protein-2 is a highly potent osteoinductive protein, which induces bone formation in vivo and osteogenic differentiation in vitro. The exogenous regulation of BMP-2 expression in bone-regenerating sites is required to control BMP-2 protein secretion, thus promoting safe and controlled bone formation and regeneration. We have therefore constructed a dual-construct vector for the recombinant AAV (rAAV)-based recombinant human BMP-2 (rhBMP-2) gene delivery system, which is regulated by the tetracycline-sensitive promoter (TetON). Each vector was encapsidated separately, yielding two recombinant viruses. We evaluated the efficiency of rAAV-hBMP-2 to induce bone formation in ectopic and orthotopic sites. Doxycycline (Dox), an analogue of tetracycline, was orally administered to mice via their drinking water to induce rhBMP-2 expression. Bone formation was measured using quantitative imaging-microcomputerized tomography and cooled charge-coupled device imaging-to detect osteogenic activity at the cellular level, detecting osteocalcin expression. The rAAV-hBMP-2-treated mice that were given Dox demonstrated bone formation in both in vivo models compared to none in mice prevented from receiving Dox. Thus, the Tet-regulated rAAV-hBMP-2 vector is an effective means of induction and regulation of bone regeneration and repair.