RESUMEN
3D cell culture systems based on biological scaffold materials obtainable from both animal and human tissues constitute very interesting tools for cell therapy and personalised medicine applications. The white adipose tissue (AT) extracellular matrix (ECM) is a very promising biomaterial for tissue engineering due to its easy accessibility, malleability and proven biological activity. In the present study, human dental pulp stem cells (hDPSCs) were combined in vitro with ECM scaffolds from porcine and human decellularised adipose tissues (pDAT, hDAT) processed as 3D solid foams, to investigate their effects on the osteogenic differentiation capacity and bone matrix production of hDPSCs, compared to single-protein-based 3D solid foams of collagen type I and conventional 2D tissue-culture-treated polystyrene plates. pDAT solid foams supported the osteogenic differentiation of hDPSCs to similar levels to collagen type I, as assessed by alkaline phosphatase and alizarin red stainings, reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) and osteocalcin/bone gamma-carboxyglutamate protein (BGLAP) immunostaining. Interestingly, hDAT solid foams showed a markedly lower capacity to sustain hDPSC osteogenic differentiation and matrix calcification and a higher capacity to support adipogenesis, as assessed by RT-qPCR and oil red O staining. White ATs from both human and porcine origins are relatively abundant and available sources of raw material to obtain high quality ECM-derived biomedical products. These biomaterials could have promising applications in tissue engineering and personalised clinical therapy for the healing and regeneration of lesions involving not only a loss of calcified bone but also its associated soft non-calcified tissues.
Asunto(s)
Colágeno Tipo I , Osteogénesis , Tejido Adiposo , Animales , Diferenciación Celular , Células Cultivadas , Pulpa Dental , Humanos , Células Madre , Porcinos , Ingeniería de Tejidos , Andamios del TejidoRESUMEN
The conversion of healthy stem cells into cancer stem cells (CSCs) is believed to underlie tumor relapse after surgical removal and fuel tumor growth and invasiveness. CSCs often arise from the malignant transformation of resident multipotent stem cells, which are present in most human tissues. Some organs, such as the gut and the brain, can give rise to very aggressive types of cancers, contrary to the dental pulp, which is a tissue with a very remarkable resistance to oncogenesis. In this review, we focus on the similarities and differences between gut, brain and dental pulp stem cells and their related CSCs, placing a particular emphasis on both their shared and distinctive cell markers, including the expression of pluripotency core factors. We discuss some of their similarities and differences with regard to oncogenic signaling, telomerase activity and their intrinsic propensity to degenerate to CSCs. We also explore the characteristics of the events and mutations leading to malignant transformation in each case. Importantly, healthy dental pulp stem cells (DPSCs) share a great deal of features with many of the so far reported CSC phenotypes found in malignant neoplasms. However, there exist literally no reports about the contribution of DPSCs to malignant tumors. This raises the question about the particularities of the dental pulp and what specific barriers to malignancy might be present in the case of this tissue. These notable differences warrant further research to decipher the singular properties of DPSCs that make them resistant to transformation, and to unravel new therapeutic targets to treat deadly tumors.
RESUMEN
Dental implants are the usual therapy of choice in the dental clinic to replace a loss of natural teeth. Over recent decades there has been an important progress in the design and manufacturing of titanium implant surfaces with the goal of improving their osteointegration. In the present work, the aim was to evaluate the usefulness of hDPSCs (human dental pulp stem cells), in combination with autologous plasma components, for in vitro bone generation on biomimetic titanium dental implant materials. In this context, the combination of hDPSCs stimulated by PRGF or PRF and cultured on standard Ti6A14V and biomimetic BAS™ (Avinent Implant System) titanium surfaces were studied in order to evaluate possible enhancements in the osteoblastic differentiation process out of human mesenchymal cells, as well as bone matrix secretion on the implant surface. The results obtained in this in vitro model of osteogenesis suggested a combination of biomimetic rough titanium surfaces, such as BAS™, with autologous plasma-derived fibrin-clot membranes such as PRF and/or insoluble PRGF formulations, but not with an addition of water-soluble supplements of plasma-derived growth factors, to maximise osteoblastic cell differentiation, bone generation, anchorage and osteointegration of titanium-made dental implants.
Asunto(s)
Materiales Biomiméticos/química , Adhesión Celular , Pulpa Dental/citología , Células Madre Mesenquimatosas/efectos de los fármacos , Oseointegración , Osteogénesis , Plasma Rico en Plaquetas/química , Adulto , Materiales Biomiméticos/farmacología , Células Cultivadas , Implantes Dentales , Fibrina/farmacología , Humanos , Péptidos y Proteínas de Señalización Intercelular/farmacología , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/fisiología , Osteoblastos/citología , Osteoblastos/efectos de los fármacos , Osteoblastos/fisiología , Titanio/químicaRESUMEN
In the original publication of the article, some symbols in Figure 3 were not correctly aligned with the image.
RESUMEN
DLK1 and DLK2 are transmembrane proteins belonging to the EGF-like repeat-containing family that function as non-canonical NOTCH inhibitory ligands. DLK1 is usually downregulated after embryo development and its distribution in some adult and embryonic tissues has been described. However, the expression and role of DLK2 in embryo and adult tissues remains unclear. To better understand the relevance of both proteins during embryo development, we analyzed the expression pattern of DLK1 and DLK2 in 16.5-day-old mouse embryos (E16.5) and evaluated the possible relationship between these two proteins in embryo tissues and cell types. We found that DLK1 and DLK2 proteins exhibited a broad distribution pattern, which was detected in developing mouse organs from each of the three germ layers: ectoderm (brain, salivary glands), mesoderm (skeletal muscle, vertebral column, kidney, cartilage), and endoderm (thymus, lung, pancreas, intestine, liver). The expression pattern of DLK1 and DLK2 indicates that both proteins could play a synergistic role during cell differentiation. This study provides additional information for understanding temporal and site-specific effects of DLK1 and DLK2 during embryo morphogenesis and cell differentiation.
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Desarrollo Embrionario , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Animales , Proteínas de Unión al Calcio , Diferenciación Celular , RatonesRESUMEN
Dental pulp stem cells (DPSCs) from adult teeth express neural crest (NC) markers together with core transcriptional factors associated with stem cell pluripotency, such as Oct4a, Sox2, c-Myc, Rex1, Stella/Dppa3, Ssea1/Fut4, Lin28 and Nanog. The possibility to boost the natural stemness features of DPSCs by mild methods, that do not involve gene and/or chromatin modification or gene transfection, is highly desirable for cell therapy. Canonical Wnt and Notch are two highly conserved developmental signalling pathways that are involved in NC emergence and stem cell self-renewal. We determined that both pathways coordinate to regulate the expression of core pluripotency and NC factors in DPSCs. Pharmacological inhibition of the Notch pathway for 48 h, by the γ-secretase inhibitor N-[N-(3,5-Difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester (DAPT), abolished the expression of NC and core factors. In addition, it induced a silencing of the canonical Wnt signalling and a clear reduction in the stemness potential of DPSCs, as shown by a reduced ability to generate mature, fully differentiated osteoblasts and adipocytes. Conversely, pharmacological activation of the Wnt pathway for 48 h, by either the glycogen synthase kinase 3 beta (GSK3-ß) inhibitor 6-bromoindirubin-3´-oxime (BIO) or the human recombinant protein Wnt-3a, not only largely increased the expression of NC and core factors, but also increased the efficiency of DPSCs to differentiate into mature osteoblasts and adipocytes. These results showed that a short preconditioning activation of Wnt/Notch signalling by small molecules and/or recombinant proteins enhanced the stemness and potency of DPSCs in culture, which could be useful for optimising the therapeutic use of these and other tissue-specific stem cells.
Asunto(s)
Autorrenovación de las Células/genética , Expresión Génica , Cresta Neural/metabolismo , Células Madre Pluripotentes/metabolismo , Transducción de Señal/genética , Células Madre/metabolismo , Adolescente , Adulto , Células Cultivadas , Pulpa Dental/citología , Dipéptidos/farmacología , Humanos , Interferencia de ARN , Receptores Notch/genética , Transducción de Señal/efectos de los fármacos , Factores de Transcripción/genética , Vía de Señalización Wnt/efectos de los fármacos , Vía de Señalización Wnt/genéticaRESUMEN
DLK1 (PREF1, pG2, or FA1) is a transmembrane and secreted protein containing epidermal growth factor-like repeats. Dlk1 expression is abundant in many tissues during embryonic and fetal development and is believed to play an important role in the regulation of tissue differentiation and fetal growth. After birth, Dlk1 expression is abolished in most tissues but is possibly reactivated to regulate stem cell activation and responses to injury. We have recently reported that DLK1 regulates many aspects of salivary gland organogenesis. Here, we have extended our studies of the salivary gland phenotype of Dlk1 knock-out mice. We have observed that salivary glands are smaller and weigh significantly less in both Dlk1 knock-out males and females compared with gender and age-matched wild-type mice and regardless of the natural sexual dimorphism in rodent salivary glands. This reduced size correlates with a reduced capacity of Dlk1-deficient mice to secrete saliva after stimulation with pilocarpine. However, histological and ultrastructural analyses of both adult and developing salivary gland tissues have revealed no defects in Dlk1 ((-/-)) mice, indicating that genetic compensation accounts for the relatively mild salivary phenotype in these animals. Finally, despite their lack of severe anomalies, we have found that salivary glands from Dlk1-deficient mice present a higher amount of CK14-positive epithelial progenitors at various developmental stages, suggesting a role for DLK1 in the regulation of salivary epithelial stem cell balance.
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Células Epiteliales/metabolismo , Células Epiteliales/patología , Péptidos y Proteínas de Señalización Intercelular/deficiencia , Glándulas Salivales/patología , Células Madre/metabolismo , Animales , Animales Recién Nacidos , Biomarcadores/metabolismo , Proteínas de Unión al Calcio , Femenino , Ganglios Parasimpáticos/metabolismo , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Queratina-14/metabolismo , Queratina-5/metabolismo , Masculino , Ratones Endogámicos C57BL , Ratones Noqueados , Tamaño de los Órganos , Saliva , Glándulas Salivales/embriología , Glándulas Salivales/inervación , Glándulas Salivales/ultraestructura , Regulación hacia ArribaRESUMEN
Redesigning lignin, the aromatic polymer fortifying plant cell walls, to be more amenable to chemical depolymerization can lower the energy required for industrial processing. We have engineered poplar trees to introduce ester linkages into the lignin polymer backbone by augmenting the monomer pool with monolignol ferulate conjugates. Herein, we describe the isolation of a transferase gene capable of forming these conjugates and its xylem-specific introduction into poplar. Enzyme kinetics, in planta expression, lignin structural analysis, and improved cell wall digestibility after mild alkaline pretreatment demonstrate that these trees produce the monolignol ferulate conjugates, export them to the wall, and use them during lignification. Tailoring plants to use such conjugates during cell wall biosynthesis is a promising way to produce plants that are designed for deconstruction.
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Aciltransferasas/química , Aciltransferasas/genética , Lignina/química , Lignina/metabolismo , Populus/genética , Populus/metabolismo , Aciltransferasas/aislamiento & purificación , Angelica sinensis/enzimología , Angelica sinensis/genética , Pared Celular/química , Pared Celular/metabolismo , Ácidos Cumáricos/metabolismo , Genes de Plantas , Estructura Molecular , Raíces de Plantas/enzimología , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/crecimiento & desarrollo , Populus/crecimiento & desarrollo , Árboles/genética , Árboles/metabolismoAsunto(s)
Glucógeno Sintasa Quinasa 3/antagonistas & inhibidores , Odontogénesis/efectos de los fármacos , Germen Dentario/efectos de los fármacos , Vía de Señalización Wnt/efectos de los fármacos , Ameloblastos/efectos de los fármacos , Amelogenina/análisis , Animales , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Órgano del Esmalte/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Proteínas de Filamentos Intermediarios/análisis , Cloruro de Litio/farmacología , Factor de Unión 1 al Potenciador Linfoide/análisis , Mesodermo/efectos de los fármacos , Ratones , Diente Molar/efectos de los fármacos , Diente Molar/embriología , Proteínas del Tejido Nervioso/análisis , Nestina , Odontoblastos/efectos de los fármacos , Técnicas de Cultivo de Órganos , Calcificación de Dientes/efectos de los fármacos , Corona del Diente/efectos de los fármacos , Corona del Diente/embriología , Raíz del Diente/efectos de los fármacos , Raíz del Diente/embriologíaAsunto(s)
Ameloblastos/patología , Claudina-1/análisis , Factores de Transcripción de Tipo Kruppel/genética , Odontoblastos/patología , Dedos de Zinc/genética , Proteína de la Zonula Occludens-1/análisis , Animales , Diferenciación Celular/fisiología , Membrana Celular/patología , Núcleo Celular/patología , Polaridad Celular/fisiología , Proteínas de Unión al ADN/análisis , Esmalte Dental/anomalías , Esmalte Dental/patología , Epitelio/patología , Incisivo/patología , Ratones , Ratones Noqueados , Diente Molar/patología , Diente Supernumerario/genética , Diente Supernumerario/patología , Factores de Transcripción/análisisRESUMEN
Post-eruptive loss of ameloblasts requires identification of alternative sources for these cells to realize tooth-tissue-engineering strategies. Recent reports showed that bone-marrow-derived cells can give rise to different types of epithelial cells, suggesting their potential to serve as a source for ameloblasts. To investigate this potential, we mixed c-Kit(+)-enriched bone marrow cells with embryonic dental epithelial cells and cultured them in re-association with dental mesenchyme. Non-dividing, polarized, and secretory ameloblast-like cells were achieved without cell fusion. Before basement membrane reconstitution, some bone marrow cells migrated to the mesenchyme, where they exhibited morphological, molecular, and functional characteristics of odontoblasts. These results show, for the first time, that bone-marrow-derived cells can be reprogrammed to give rise to ameloblast-like cells, offering novel possibilities for tooth-tissue engineering and the study of the simultaneous differentiation of one bone marrow cell subpopulation into cells of two different embryonic lineages.
Asunto(s)
Ameloblastos/citología , Células de la Médula Ósea/citología , Ingeniería de Tejidos/métodos , Animales , Diferenciación Celular , Proliferación Celular , Células Cultivadas , Medios de Cultivo Condicionados , Femenino , Técnica del Anticuerpo Fluorescente , Hibridación in Situ , Mesodermo/citología , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos , Proteínas Proto-Oncogénicas c-kit/fisiologíaRESUMEN
Tight junctions might play a role during tissue morphogenesis and cell differentiation. In order to address these questions, we have studied the distribution pattern of the tight junction-associated proteins ZO-1, ZO-2, ZO-3 and occludin in the developing mouse tooth as a model. A specific temporal and spatial distribution of tight junction-associated proteins during tooth development was observed. ZO-1 appeared discontinuously in the cell membrane of enamel organ and dental mesenchyme cells. However, endothelial cells of the dental mesenchyme capillaries displayed a continuous fluorescence at the cell membrane. Inner dental epithelium first showed an evident signal for ZO-1 at the basal pole of the cells at bud/cap stage, but ZO-1 was accumulated at the basal and apical pole of preameloblast/ameloblasts at late bell stage. Surprisingly, in the incisor ZO-1 decreased as the inner dental epithelium differentiated, and was re-expressed in secretory and mature ameloblasts. On the contrary, ZO-2 was confined to continuous cell-cell contacts of the enamel organ in both molars and incisors. The lateral cell membrane of inner dental epithelial cells was specifically ZO-2 labeled. However, ZO-3 was expressed in oral epithelium whereas dental embryo tissues were negative. In addition, occludin was hardly detected in dental tissues at the early stage of tooth development, but was distributed continuously at the cell membrane of endothelial cells of ED19.5 dental mesenchyme. In incisors, occludin was detected at the cell membrane of the secretory pole of ameloblasts. The occurrence and relation during tooth development of tight junction proteins ZO-1, ZO-2 and occludin, but not ZO-3, suggests a combinatory assembly in tooth morphogenesis and cell differentiation.
Asunto(s)
Proteínas Portadoras/biosíntesis , Proteínas de la Membrana/biosíntesis , Fosfoproteínas/biosíntesis , Diente/embriología , Actinas/metabolismo , Animales , Immunoblotting , Ratones , Ocludina , Uniones Estrechas/metabolismo , Diente/metabolismo , Proteínas de la Zonula Occludens , Proteína de la Zonula Occludens-1 , Proteína de la Zonula Occludens-2RESUMEN
Signaling molecules, their receptors, and target genes from pathways and networks regulate the development of the tooth from initiation through cell differentiation. In order to identify genes involved in odontoblast and ameloblast differentiation, we constructed a cDNA library from E19.5 mouse molars. In this work, we report the partial cDNA sequences of 10 noncharacterized genes and we show cell expression of the transcripts on mouse embryo molars by in situ hybridization.
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ADN Complementario/genética , Ratones/embriología , Ratones/genética , Diente Molar/embriología , Odontogénesis/genética , Animales , ADN Complementario/metabolismo , Embrión de Mamíferos/metabolismo , Biblioteca de Genes , ARN Mensajero/metabolismo , Homología de Secuencia de AminoácidoRESUMEN
We have studied the expression of FGF1 and FGF2 during mouse odontogenesis by immunohistochemistry. FGF1 was detected in differentiated odontoblasts and at the secretory pole of ameloblasts. Localization of FGF2 was mainly observed within the basement membrane interposed between dental epithelium and dental mesenchyme. These findings indicate that FGF1 and FGF2 may participate in the control of odontoblast and ameloblast differentiation. Thereafter, we studied the ability of FGF1 and FGF2, alone or in combination with TGF beta 1, to induce polarization and/or functional differentiation of preodontoblasts. Dental papillae (DP) obtained from first lower molars of 17-day-old mouse embryo were cultured in the presence or the absence of growth factors. DP cultured with FGF1 + TGF beta 1 showed gradients of odontoblast-like cell differentiation, which displayed alkaline phosphatase reactivity. DP treated with FGF2 + TGF beta 1 exhibited pre-odontoblast cell polarization, and the cell bodies displayed long cytoplasm processes. However, following this treatment we did not observe extracellular matrix secretion, and alkaline phosphatase activity was completely inhibited. In summary, our results show that exogenous addition of FGF1 to pre-odontoblasts induces their terminal differentiation, by synergistically acting with TGF beta 1. In contrast, FGF2 may regulate the effect of TGF beta 1, permitting cell polarization but restraining pre-odontoblast functions.
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Factor 1 de Crecimiento de Fibroblastos/farmacología , Factor 2 de Crecimiento de Fibroblastos/farmacología , Odontoblastos/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Factor de Crecimiento Transformador beta/farmacología , Fosfatasa Alcalina/análisis , Ameloblastos/efectos de los fármacos , Ameloblastos/metabolismo , Animales , Membrana Basal/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Polaridad Celular/efectos de los fármacos , Técnicas de Cultivo , Citoplasma/efectos de los fármacos , Citoplasma/ultraestructura , Papila Dental/citología , Papila Dental/efectos de los fármacos , Epitelio/efectos de los fármacos , Matriz Extracelular/efectos de los fármacos , Matriz Extracelular/metabolismo , Inmunohistoquímica , Mesodermo/efectos de los fármacos , Ratones , Microscopía Electrónica , Odontogénesis/efectos de los fármacos , Factor de Crecimiento Transformador beta1RESUMEN
Psoriasis is most probably an inherited disease characterized by cell proliferation, angiogenesis, and an inflammatory process. The pathophysiology remains unknown, although an alteration in cell-cell and cell-matrix adhesion versus an autoimmune process has been proposed as the primary defect. Here, we show evidence of a new mechanism involving basement membrane alterations accompanied by keratinocyte overexpression of matrix metalloproteinase (MMP) 2 and tissue inhibitor of MMP-2 (TIMP-2) in both uninvolved and involved psoriatic skin. Immunocytochemistry with antibodies against collagen IV (alpha1, alpha2 chains) and laminins (alpha2, alpha5, beta1, gamma1 chains) revealed gaps, folding, and reduplication of the epidermo-dermal basement membrane. There was overexpression of MMP-2 in the cytoplasm of suprabasal keratinocytes. Gelatin zymography revealed pro-MMP-2 and its activated form, a-MMP-2, in both uninvolved and involved psoriatic skin, whereas pro-MMP-9 was only present in involved skin. TIMP-2 was expressed at the cell surface of psoriatic involved suprabasal keratinocytes whereas it was restricted to basal keratinocytes in uninvolved areas. Western blots showed a marked increase in a-MMP-2 and TIMP-2 in uninvolved and involved psoriatic skin although it was more pronounced in the latter. MT1-MP, known to activate pro-MMP-2, was increased in involved areas. In situ hybridization revealed strong signals of MMP-2 mRNA in both uninvolved and involved psoriatic epidermis. The overexpression of MMP-2 in uninvolved and involved psoriatic epidermis supports the concept that the primary alteration may reside in the keratinocyte. In addition, the presence of the activated form of MMP-2 could be responsible for cell-cell and cell-matrix changes noted in psoriatic epidermis.
Asunto(s)
Membrana Basal/metabolismo , Metaloproteinasa 2 de la Matriz/biosíntesis , Psoriasis/metabolismo , Piel/química , Piel/metabolismo , Inhibidor Tisular de Metaloproteinasa-2/biosíntesis , Western Blotting , Humanos , Inmunohistoquímica , Hibridación in Situ , Metaloproteinasa 2 de la Matriz/genética , ARN Mensajero/metabolismoRESUMEN
Dental papillae (DP) isolated from first lower molars of 17-day-old mouse embryos were cultured in the presence of combinations of the following growth factors: FGF1, FGF2, and TGFbeta1. After 6 days in culture, only the DP treated with FGF1+TGFbeta1 contained differentiated odontoblast-like cells at the periphery of the explants, and these cells secreted extracellular matrix similar to predentin. Surprisingly, treatments with FGF2+TGFbeta1 induced cell polarization at the surface of the explants but no matrix secretion was observed. Electron microscopy and histochemical analysis of odontoblast markers showed that differentiated cells induced by FGF1+TGFbeta1 exhibited cytological features of functional odontoblasts with matrix vesicle secretion and mineral formation, positive alkaline-phosphatase activity, and type-I collagen production. DP cultured in the presence of FGF2+TGFbeta1 showed cell polarization and long and thin cell processes containing matrix vesicles; however, type-I collagen secretion was not detected and alkaline-phosphatase activity was completely inhibited. Our results indicate that, in our culture system, exogenous combinations of FGF1, FGF2, and TGFbeta1 interact with preodontoblasts and induce cell polarization or differentiation, which can be studied separately in vitro. Thus, FGF1 and TGFbeta1 do have a synergic effect to promote morphological and functional features of differentiated odontoblasts whereas FGF2 seems to modulate TGFbeta1 action, causing morphological polarization of preodontoblasts but limiting the functional activity of these cells in terms of type-I collagen secretion and alkaline-phosphatase activity.
Asunto(s)
Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/fisiología , Factor 2 de Crecimiento de Fibroblastos/farmacología , Odontoblastos/citología , Odontoblastos/metabolismo , Factor de Crecimiento Transformador beta/farmacología , Fosfatasa Alcalina/metabolismo , Animales , Polaridad Celular/efectos de los fármacos , Polaridad Celular/fisiología , Células Cultivadas , Colágeno/inmunología , Colágeno/metabolismo , Papila Dental/fisiología , Papila Dental/ultraestructura , Factor 1 de Crecimiento de Fibroblastos , Técnica del Anticuerpo Fluorescente , Ratones , Odontoblastos/efectos de los fármacos , ConejosRESUMEN
In this work, we investigated the effects of aFGF and bFGF alone or combined with TGFbeta1 or IGF-I on odontoblast differentiation. Trypsin-isolated dental papillae from day 17 mandibular first molar were cultured in semisolid-agar medium for 6 d. Our results demonstrated that aFGF, bFGF or combinations of these promoted cell polarization at the periphery of the dental papillae. Moreover, simultaneous addition of aFGF and TGFbeta1 to dental papillae cultures induced both polarization and functional differentiation of odontoblast-like cells, as well as extracellular matrix deposition. Combination of aFGF or bFGF with IGF-I caused cell polarization at the surface of dental papillae, but matrix secretion was restricted to a few explants. In the presence of bFGF and TGFbeta1, the explants had pronounced cell elongations but no matrix deposition. These results indicate that aFGF or bFGF is not able to induce odontoblast differentiation alone. However, both aFGF and bFGF can act synergistically with TGFbeta1 and IGF-I to strengthen their inductive effects and promote gradients of cytological and functional changes in odontoblast-like cells.
Asunto(s)
Sustancias de Crecimiento/farmacología , Odontoblastos/citología , Odontoblastos/efectos de los fármacos , Animales , Diferenciación Celular/efectos de los fármacos , Polaridad Celular/efectos de los fármacos , Papila Dental/efectos de los fármacos , Papila Dental/embriología , Papila Dental/metabolismo , Sinergismo Farmacológico , Matriz Extracelular/efectos de los fármacos , Matriz Extracelular/metabolismo , Factor 1 de Crecimiento de Fibroblastos/administración & dosificación , Factor 1 de Crecimiento de Fibroblastos/farmacología , Factor 2 de Crecimiento de Fibroblastos/administración & dosificación , Factor 2 de Crecimiento de Fibroblastos/farmacología , Sustancias de Crecimiento/administración & dosificación , Técnicas In Vitro , Factor I del Crecimiento Similar a la Insulina/administración & dosificación , Factor I del Crecimiento Similar a la Insulina/farmacología , Ratones , Odontoblastos/metabolismo , Odontogénesis/efectos de los fármacos , Odontogénesis/fisiología , Factor de Crecimiento Transformador beta/administración & dosificación , Factor de Crecimiento Transformador beta/farmacologíaRESUMEN
The existence of cell surface-associated molecules has been claimed to play a major role in cellular recognition and interaction. In this respect, different tumor cell lines express laminin and its receptor, and this expression has been correlated with metastatic potential. In the present work, we have studied, by electron microscopic immunolabeling methods, the presence of laminin and 67KDa laminin-receptor on the surface of endothelial cells of lung blood capillaries. To label these molecules, we have developed an easy method in which the labeling is carried out "in situ", in previously excised lungs. The presence of both molecules was observed on the luminal surface of endothelial capillaries and, in many cases, gold particles were associated to small open vesicles of the endothelial cells. The results suggest that these molecules, traditionally associated to extracellular matrix, are also expressed in cellular surface of the lung vascular bed.