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Data sources The electronic databases Medline, the Cochrane Central Register of Controlled Trials, Embase, China National Knowledge Infrastructure, China Science and Technology Journal Database, Wanfang Data, ClinicalTrials.gov and WHO International Clinical Trials Registry Platform, from inception to September 2020, were searched to identify the eligible studies measuring the association between periodontal disease and Alzheimer's disease or mild cognitive impairment.Study selection Cohort, cross-sectional and case-control studies, without any language restrictions, were selected by two reviewers independently.Data extraction and synthesis Data extraction and quality assessment were performed by two reviewers independently. Data was synthesised quantitatively with meta-analyses using a random or fixed-effects model, with P <0.1 considered statistically significant. Quality assessment of cohort and case-control studies was carried out using the Newcastle-Ottawa scale (NOS) and quality assessment of cross-sectional studies was undertaken using the Agency for Healthcare Research and Quality (AHRQ) tool. Heterogeneity of included studies was assessed with I2.Results Thirteen studies, including five cross-sectional studies, five case-control studies, two retrospective cohort studies and one prospective cohort study were found to be eligible. Meta-analyses showed elevated risk for Alzheimer's disease (odds ratio = 1.78; random-effects model; significant heterogeneity) and mild cognitive impairment (odds ratio = 1.60; fixed-effects model; low heterogeneity) in patients with periodontal disease. One case-control study and all cohort studies had high quality, while four case-control studies had medium quality, as evaluated by the NOS. Among the cross-sectional studies evaluated by the AHRQ tool, only one had high quality, whereas other studies had medium quality.Conclusions Within the limitations of the included studies, the authors concluded that periodontal disease is related to an elevated risk of Alzheimer's disease and mild cognitive impairment.
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Enfermedad de Alzheimer , Disfunción Cognitiva , Enfermedades Periodontales , Enfermedad de Alzheimer/complicaciones , Estudios de Casos y Controles , Estudios Transversales , Progresión de la Enfermedad , Humanos , Enfermedades Periodontales/complicaciones , Estudios Prospectivos , Estudios Retrospectivos , Sensibilidad y Especificidad , Estados UnidosRESUMEN
In clinical practice, the metabolic syndrome can lead to multiple complications, including diabetes. It remains unclear which component of the metabolic syndrome (obesity, inflammation, hyperglycemia, or insulin resistance) has the strongest inhibitory effect on stem cells involved in beta cell regeneration. This makes it challenging to develop effective treatment options for complications such as diabetes. In our study, experiments were performed on male C57BL/6 mice where metabolic disorders have been introduced experimentally by a combination of streptozotocin-treatment and a high-fat diet. We evaluated the biological effects of Bisamide Derivative of Dicarboxylic Acid (BDDA) and its impact on pancreatic stem cells in vivo. To assess the impact of BDDA, we applied a combination of histological and biochemical methods along with a cytometric analysis of stem cell and progenitor cell markers. We show that in mice with metabolic disorders, BDDA has a positive effect on lipid and glucose metabolism. The pancreatic restoration was associated with a decrease of the inhibitory effects of inflammation and obesity factors on pancreatic stem cells. Our data shows that BDDA increases the number of pancreatic stem cells. Thus, BDDA could be used as a new compound for treating complication of the metabolic syndrome such as diabetes.
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Amidas/química , Citocinas/sangre , Ácidos Dicarboxílicos/química , Hipoglucemiantes/farmacología , Lípidos/sangre , Enfermedades Metabólicas/tratamiento farmacológico , Animales , Hipoglucemiantes/química , Masculino , Enfermedades Metabólicas/sangre , Enfermedades Metabólicas/patología , Ratones , Ratones Endogámicos C57BLRESUMEN
In clinical practice, there are patients with a combination of metabolic syndrome (MS) and chronic obstructive pulmonary disease (COPD). The pathological mechanisms linking MS and COPD are largely unknown. It remains unclear whether the effect of MS (possible obesity) has a major impact on the progression of COPD. This complicates the development of effective approaches for the treatment of patients with a diagnosis of MS and COPD. Experiments were performed on female C57BL/6 mice. Introduction of monosodium glutamate and extract of cigarette smoke was modeled to simulate the combined pathology of lipid disorders and emphysema. Biological effects of glucagon-like peptide 1 (GLP-1) and GLP-1 on endothelial progenitor cells (EPC) in vitro and in vivo were evaluated. Histological, immunohistochemical methods, biochemical methods, cytometric analysis of markers identifying EPC were used in the study. The CD31⺠endothelial cells in vitro evaluation was produced by Flow Cytometry and Image Processing of each well with a Cytation™ 3. GLP-1 reduces the area of emphysema and increases the number of CD31⺠endothelial cells in the lungs of mice in conditions of dyslipidemia and damage to alveolar tissue of cigarette smoke extract. The regenerative effects of GLP-1 are caused by a decrease in inflammation, a positive effect on lipid metabolism and glucose metabolism. EPC are proposed as pathogenetic and diagnostic markers of endothelial disorders in combination of MS with COPD. Based on GLP-1, it is proposed to create a drug to stimulate the regeneration of endothelium damaged in MS and COPD.
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Péptido 1 Similar al Glucagón/genética , Síndrome Metabólico/genética , Enfermedad Pulmonar Obstructiva Crónica/genética , Enfisema Pulmonar/genética , Animales , Fumar Cigarrillos/efectos adversos , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Células Progenitoras Endoteliales/metabolismo , Citometría de Flujo , Glucosa/metabolismo , Humanos , Metabolismo de los Lípidos/genética , Pulmón/efectos de los fármacos , Pulmón/patología , Síndrome Metabólico/inducido químicamente , Síndrome Metabólico/diagnóstico , Síndrome Metabólico/tratamiento farmacológico , Ratones , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/genética , Enfermedad Pulmonar Obstructiva Crónica/inducido químicamente , Enfermedad Pulmonar Obstructiva Crónica/diagnóstico , Enfermedad Pulmonar Obstructiva Crónica/tratamiento farmacológico , Enfisema Pulmonar/diagnóstico , Enfisema Pulmonar/tratamiento farmacológico , Enfisema Pulmonar/patología , Glutamato de Sodio/toxicidadRESUMEN
The aim of this study was to analyze the success rate of dental implants and the graft shrinkage rate after vertical ridge augmentation and simultaneous implantation with an allograft bonering. Fifty-one patients (81 augmentations and simultaneous implantations) were included. The bonering technique followed a standardized protocol. The alveolar ridge was prepared using a congruent trephine, and depending on the defect size, an allograft bonering with an outer diameter of 6-7 mm was placed. The height of the bonering was trimmed with a diamond disc to the required length. The average height of vertical augmentation was 5.5 mm. Implants were inserted through the bonering into the native bone of alveolar ridge. After 6 months, dental implants were exposed, and dental prosthetics were placed. Of 81 implants placed with the bonering technique, two failed during a 12-month follow-up, corresponding to a success rate of 97.5%. One year after surgery, the allograft bonering exhibited an average vertical graft shrinkage rate of 8.6%. In conclusion, the allograft bonering technique was associated with a favorable outcome, and in cases with large vertical defects, both treatment time and donor site morbidity could be reduced.
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Aumento de la Cresta Alveolar , Implantes Dentales , Aloinjertos , Trasplante Óseo , Implantación Dental , Implantación Dental Endoósea , Humanos , Resultado del TratamientoRESUMEN
Idiopathic pulmonary fibrosis (IPF) is one of the most common forms of interstitial lung disease, characterized by progressive parenchymal fibrosis and respiratory failure. In a model of bleomycin-induced pulmonary fibrosis, the antifibrotic and anti-inflammatory activity of Longidaze (Bovhyaluronidase Azoxymer), which contains a conjugate of the hyaluronidase enzyme with a high molecular weight synthetic carrier azoxymer bromide, was investigated. Experiments were conducted in male C57BL/6 mice. Longidaze was administered at different doses by intranasal and intramuscular routes. Histology, hematology, and enzyme-linked immunosorbent assay were used in the study. The use of Longidaze reduced pulmonary fibrosis, as evidenced by an improvement in histopathologic damage to the lungs, a decrease in the area of connective tissue, and the levels of profibrotic factors (TGF-ß1, hydroxyproline, collagen I) in lung tissue. In addition, Longidaze inhibited the inflammatory response in pulmonary fibrosis, and decreased the levels of IL-6, TNF-α, and hyaluronic acid in lung tissue and the recruitment of inflammatory cells into lung tissue. The highest therapeutic efficacy was observed with the use of Longidaze at doses of 120 and 1200 U/kg intramuscularly, which was superior to that of the reference drug pirfenidone axunio. The data presented in this study suggest that Longidaze is a new and promising drug for the treatment of IPF that warrants further investigation in patients with fibrotic interstitial lung disease.
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Finding new, safe strategies to prevent and control rheumatoid arthritis is an urgent task. Bioactive peptides and peptide-rich protein hydrolyzate represent a new trend in the development of functional foods and nutraceuticals. The resulting tissue hydrolyzate of the chicken embryo (CETH) has been evaluated for acute toxicity and tested against chronic arthritis induced by Freund's full adjuvant (modified Mycobacterium butyricum) in rats. The antiarthritic effect of CETH was studied on the 28th day of the experiment after 2 weeks of oral administration of CETH at doses of 60 and 120 mg/kg body weight. Arthritis was evaluated on the last day of the experiment on the injected animal paw using X-ray computerized microtomography and histopathology analysis methods. The CETH effect was compared with the non-steroidal anti-inflammatory drug diclofenac sodium (5 mg/kg). Oral administration of CETH was accompanied by effective dose-dependent correction of morphological changes caused by the adjuvant injection. CETH had relatively high recovery effects in terms of parameters for reducing inflammation, inhibition of osteolysis, reduction in the inflammatory reaction of periarticular tissues, and cartilage degeneration. This study presents for the first time that CETH may be a powerful potential nutraceutical agent or bioactive component in the treatment of rheumatoid arthritis.
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A technology to create a cell-seeded fibrin-based implant matching the size and shape of bone defect is required to create an anatomical implant. The aim of the study was to develop a technology of cell-seeded fibrin gel implant creation that has the same shape and size as the bone defect at the site of implantation. Using computed tomography (CT) images, molds representing bone defects were created by 3D printing. The form was filled with fibrin glue and human dental pulp stem cells (DPSC). The viability, set of surface markers and osteogenic differentiation of DPSC grown in fibrin gel along with the clot retraction time were evaluated. In mice, an alveolar bone defect was created. The defect was filled with fibrin gel seeded with mouse DPSC. After 28 days, the bone repair was analyzed with cone beam CT and by histological examination. The proliferation rate, set of surface antigens and osteogenic potential of cells grown inside the scaffold and in 2D conditions did not differ. In mice, both cell-free and mouse DPSC-seeded implants increased the bone tissue volume and vascularization. In mice with cell-seeded gel implants, the bone remodeling process was more prominent than in animals with a cell-free implant. The technology of 3D-printed forms for molding implants can be used to prepare implants using components that are not suitable for 3D printing.
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In food biotechnology, Pleurotus ostreatus is of great interest as a source of natural antioxidants and angiotensin-converting enzyme (ACE) inhibitors. However, research in this area has not yet been completed. The effect of various drying methods on the structural properties and the rehydration capacity of mushrooms was investigated in this paper. The content of secondary metabolites, the peptide profile, and the antioxidative effect and ACE inhibitory activity of dry mushrooms were investigated in vitro, simulating the process of gastrointestinal digestion. X-ray microtomography has confirmed that structure of lyophilic and sun-dried mushrooms is dominated by open pores, and in mushrooms dried with hot air and microwave, closed pores. Experiments have shown that the conditions of freeze drying and sun drying of Pleurotus ostreatus provide a higher rehydration capacity of dried mushrooms. The maximum activity of radical absorption of the oyster mushroom after microwave drying was observed. The iron restoring capacity of the mushrooms is maximally maintained with microwave drying and hot-air drying. The properties of the antioxidant product with an emphasis on the high activity of inhibiting lipid oxidation of the mushroom maximized after drying in the sun. Mushrooms dried lyophilically and in the sun showed the highest ACE inhibitory activity.
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BACKGROUND: Enamel matrix derivative (EMD) has a low immunogenic potential. To the best of our knowledge, there are no studies on the influence of EMD on lymphocyte migration as a sensitive cellular reaction parameter. This study investigated the influence of EMD on primary T-lymphocyte migration, CD25 activation, and activation-induced cell death. METHODS: After immunomagnetic-positive CD4+ lymphocyte separation from peripheral blood taken from three healthy volunteers per trial, the influence of EMD on cell locomotion was assessed in a three-dimensional collagen matrix migration model (CMMM). Direct CD4+ cell contact with EMD at concentrations of 25 and 100 microg/ml was mediated in a one-phase CMMM. We investigated the indirect influence of EMD in a two-phase CMMM: one collagen phase contained 25 and 100 microg EMD/ml, using the same concentrations, and a second adjacent phase contained T lymphocytes. After time-lapse videomicroscopy, the mean locomoting percentage of 30 randomly selected cells was analyzed. Using flow cytometry, CD25 receptor activation was assessed, and annexin V was used for apoptosis detection in lymphocytes challenged with 0, 1, 25, 50, and 100 microg EMD/ml. RESULTS: The one-phase CMMM revealed a reduction and the two-phase CMMM showed a dose-dependent increase in the mean locomoting cell percentage (P <0.001). Increasing EMD concentrations resulted in dose-dependent enhanced T-cell CD25 receptor expression and in increasing apoptosis (P <0.001). CONCLUSIONS: Our study showed immediate effects of EMD on primary CD4+ lymphocyte migration, CD25 activation, and apoptosis. CD4+ lymphocyte apoptosis may be a further possible background for uneventful early wound healing as seen clinically as the result of EMD application.
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Apoptosis/efectos de los fármacos , Quimiotaxis de Leucocito/efectos de los fármacos , Proteínas del Esmalte Dental/farmacología , Factores Inmunológicos/farmacología , Subunidad alfa del Receptor de Interleucina-2/efectos de los fármacos , Linfocitos T Colaboradores-Inductores/efectos de los fármacos , Adulto , Anexina A5 , Linfocitos T CD4-Positivos/efectos de los fármacos , Colágeno , Femenino , Citometría de Flujo , Fluoresceína-5-Isotiocianato/análogos & derivados , Colorantes Fluorescentes , Humanos , Masculino , Microscopía por Video , Persona de Mediana Edad , Adulto JovenRESUMEN
The search for osteoinductive as well as osteoconductive materials has led to the novel idea of using titanium in bone augmentations of the alveolar crest. Due to its excellent biocompatibility and favorable osteogenic properties, highly porous TiO2 granules has been proposed as a promising material for non-resorbable synthetic bone grafts in the restoration of large bone defects, and for bone augmentation in dental applications. OBJECTIVES: The aim of this study was to investigate the osteoconductive properties and biological performance of porous titanium granules used in osseous defects adjacent to the maxillary sinus in sheep. The experimental animal study involved 15 yearling sheep with a focus on the osteogenic potential of porous titanium used for subantral augmentation. MATERIAL AND METHODS: Calibrated defects were prepared in the subantral region of sheep. The defects were randomized into tests and control group. The test defects were grafted with porous titanium granules (PTG), whereas control defects were left empty (sham). Defects were left for healing for 30, 60, and 90 days. After healing, the grafted areas were removed and finally osteoconductivity was analyzed by an orthopantograph (OPG} and histology. RESULTS: Significantly more new bone formed in PTG grafted defects compared with sham. The control group showed significantly less expression of key inflammation cells, but with no significant difference in key inflammation cells compared with the experimental groups. CONCLUSION: Porous titanium can offer as an effective alternative to calcium phosphate and bone collagen-based materials used for subantral augmentation of the maxillary bone in cases of dental implantation.
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Osteogénesis , Elevación del Piso del Seno Maxilar/métodos , Titanio , Animales , Materiales Biocompatibles , Sustitutos de Huesos/uso terapéutico , Seno Maxilar/cirugía , Ovinos , Elevación del Piso del Seno Maxilar/instrumentaciónRESUMEN
Recent studies have revealed the presence of postnatal stem cells in tissues of dental origin. Our objective was to establish a standardized in vitro model system to investigate periodontal regenerative procedures for potential clinical application. We aimed to prepare primary cell cultures from human periodontal ligament and to identify clonogenic progenitor cells. After scraping PDL tissue from extracted wisdom teeth, the extracellular matrix was enzymatically degraded to obtain isolated cells for culturing. The effect of FCS and Emdogain on cell viability of the cultures was estimated by MTT-assay. Cell populations expressing STRO-1 mesenchymal, c-kit embryonic and CD34 hematopoietic stem cell markers were identified by FACS-analysis. We successfully established primary cell cultures from the human PDL. The proliferation rate of the cultures was enhanced by the supplementation of the culture medium by serum or Emdogain. The PDL cultures contained cells capable of colony-formation, as well as cells with STRO-1, c-kit and CD-34 expression. The primary cultures were maintained through multiple passages. These findings present a novel opportunity to further investigate the differentiation and proliferation of PDL derived cells potentially capable of periodontal regeneration.
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Ligamento Periodontal , Regeneración , Células Madre , Antígenos CD34/análisis , Antígenos de Superficie/análisis , Biomarcadores/análisis , Separación Celular , Tratamiento Basado en Trasplante de Células y Tejidos , Células Cultivadas , Humanos , Ligamento Periodontal/química , Ligamento Periodontal/citología , Ligamento Periodontal/fisiología , Proteínas Proto-Oncogénicas c-kit/análisisRESUMEN
Adult mammalian craniofacial tissues contain limited numbers of post-migratory neural crest-derived stem cells. Similar to their embryonic counterparts, these adult multipotent stem cells can undergo multi-lineage differentiation and are capable of contributing to regeneration of mesodermal and ectodermal cells and tissues in vivo. In the present study, we describe for the first time the presence of Nestin-positive neural crest-derived stem cells (NCSCs) within the ovine hard palate. We show that these cells can be isolated from the palatal tissue and are able to form neurospheres. Ovine NCSCs express the typical neural crest markers Slug and Twist, exhibit high proliferative and migratory activity and are able to differentiate into α smooth muscle cells and ß-III-tubulin expressing ectodermal cells. Finally, we demonstrate that oNCSCs are capable of differentiating into osteogenic, adipogenic and chondrogenic cells. Taken together, our results suggest that oNCSCs could be used as model cells to assess the efficacy and safety of autologous NCSC transplantation in a large animal model.
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Neural stem cells (NSCs) are potential sources for cell therapy of neurodegenerative diseases and for drug screening. Despite their potential benefits, ethical and practical considerations limit the application of NSCs derived from human embryonic stem cells (ES) or adult brain tissue. Thus, alternative sources are required to satisfy the criteria of ready accessibility, rapid expansion in chemically defined media and reliable induction to a neuronal fate. We isolated somatic stem cells from the human periodontium that were collected during minimally invasive periodontal access flap surgery as part of guided tissue regeneration therapy. These cells could be propagated as neurospheres in serum-free medium, which underscores their cranial neural crest cell origin. Culture in the presence of epidermal growth factor (EGF) and fibroblast growth factor-2 (FGF-2) under serum-free conditions resulted in large numbers of nestin-positive/Sox-2-positive NSCs. These periodontium-derived (pd) NSCs are highly proliferative and migrate in response to chemokines that have been described as inducing NSC migration. We used immunocytochemical techniques and RT-PCR analysis to assess neural differentiation after treatment of the expanded cells with a novel induction medium. Adherence to substrate, growth factor deprivation, and retinoic acid treatment led to the acquisition of neuronal morphology and stable expression of markers of neuronal differentiation by more than 90% of the cells. Thus, our novel method might provide nearly limitless numbers of neuronal precursors from a readily accessible autologous adult human source, which could be used as a platform for further experimental studies and has potential therapeutic implications.
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Diferenciación Celular/fisiología , Neuronas/fisiología , Periodoncio , Células Madre , Adolescente , Adulto , Biomarcadores/metabolismo , Calcio/metabolismo , Linaje de la Célula , Proliferación Celular , Células Cultivadas , Citometría de Flujo , Regeneración Tisular Guiada Periodontal , Humanos , Persona de Mediana Edad , Neuronas/citología , Periodoncio/citología , Periodoncio/cirugía , Fenotipo , Células Madre/citología , Células Madre/fisiologíaRESUMEN
BACKGROUND: Centella asiatica and Punica granatum are medicinal plants that have been reported to promote tissue healing and modulate host responses. Preliminary study revealed positive clinical effects of an innovative preparation from the two herbal extracts in the form of biodegradable chips as a subgingival adjunct to scaling and root planing. The purpose of this research was to evaluate further augmenting efficacy the combined herbal preparation may have among maintenance patients in comparison to standard supportive periodontal therapy (SPT), with additional monitoring of certain inflammatory markers. METHODS: Fifteen patients in the recall programme who had completed conventional periodontal therapy with remaining probing pocket depths of 5-8 mm were enrolled. After baseline examination and collection of gingival crevicular fluid (GCF) samples, SPT was provided and the target teeth in the test group received subgingival delivery of the medicated chips. The clinical parameters, which included probing pocket depth (PD), attachment level (AL), bleeding index (BI), gingival index (GI) and plaque index (PI) were recorded and GCF samples were collected at baseline, 3 and 6 months. RESULTS: The results showed significant improvement of PD, AL, and GI at 3 and 6 months and of BI at 6 months in the test group as compared to control. No significant differences in PI were found between the two treatment modalities at all subsequent visits. The test group also showed statistically greater reduction of IL-1beta at both 3 and 6 months and lower IL-6 concentration, which almost reached the level of significance at 6 months. CONCLUSION: The results indicate that adjunctive local delivery of extracts from C. asiatica in combination with P. granatum significantly improved clinical signs of chronic periodontitis and IL-1beta level in maintenance patients.
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Centella , Lythraceae , Periodontitis/tratamiento farmacológico , Fitoterapia , Extractos Vegetales/uso terapéutico , Adulto , Anciano , Raspado Dental , Combinación de Medicamentos , Femenino , Humanos , Interleucina-1/análisis , Interleucina-6/análisis , Masculino , Persona de Mediana Edad , Estadísticas no ParamétricasRESUMEN
The management of facial defects has rapidly changed in the last decade. Functional and esthetic requirements have steadily increased along with the refinements of surgery. In the case of advanced atrophy or jaw defects, extensive horizontal and vertical bone augmentation is often unavoidable to enable patients to be fitted with implants. Loss of vertical alveolar bone height is the most common cause for a non primary stability of dental implants in adults. At present, there is no ideal therapeutic approach to cure loss of vertical alveolar bone height and achieve optimal pre-implantological bone regeneration before dental implant placement. Recently, it has been found that specific populations of stem cells and/or progenitor cells could be isolated from different dental resources, namely the dental follicle, the dental pulp and the periodontal ligament. Our research group has cultured palatal-derived stem cells (paldSCs) as dentospheres and further differentiated into various cells of the neuronal and osteogenic lineage, thereby demonstrating their stem cell state. In this publication will be shown whether paldSCs could be differentiated into the osteogenic lineage and, if so, whether these cells are able to regenerate alveolar bone tissue in vivo in an athymic rat model. Furthermore, using these data we have started a proof of principle clinical- and histological controlled study using stem cell-rich palatal tissues for improving the vertical alveolar bone augmentation in critical size defects. The initial results of the study demonstrate the feasibility of using stem cell-mediated tissue engineering to treat alveolar bone defects in humans.
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Periodontium-derived stem cells (pdSCs) can be cultured as dentospheres and differentiated into various cells of the neuronal lineage such as glial cells, thereby demonstrating their stem cell state. This study investigated whether pdSCs could be differentiated into the osteogenic lineage and, if so, whether these cells are able to regenerate periodontal tissue in vivo in an athymic rat model. Human adult pdSCs were isolated during minimally invasive periodontal surgery and expanded in vitro. To induce osteogenic differentiation, expanded pdSCs were cultured for 3 weeks in osteogenic differentiation media. Staining for alkaline phosphatase expression was positive, suggesting osteogenic differentiation. For in vivo studies, pdSCs were delivered onto suitable collagen sponges and implanted into periodontal defects on the right buccal cortex of the mandible in 16 immunodeficient nude rats. Histologic analysis of samples from the test side revealed reformation of periodontal ligament-like tissue, collagen fibers, and elements of bone, but no functional periodontal tissue regeneration. The data show that human adult pdSCs are capable of regenerating elements of bone and collagen fibers in an in vivo animal model.
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Células Madre Adultas/fisiología , Periodoncio/citología , Regeneración/fisiología , Adulto , Fosfatasa Alcalina/análisis , Pérdida de Hueso Alveolar/cirugía , Proceso Alveolar/patología , Animales , Regeneración Ósea/fisiología , Técnicas de Cultivo de Célula , Diferenciación Celular/fisiología , Linaje de la Célula , Colágeno , Cemento Dental/patología , Dentina/patología , Humanos , Mandíbula/cirugía , Microscopía Confocal , Persona de Mediana Edad , Osteocitos/patología , Osteogénesis/fisiología , Ligamento Periodontal/patología , Periodoncio/fisiología , Distribución Aleatoria , Ratas , Ratas Desnudas , Andamios del Tejido , Adulto JovenRESUMEN
The aim of this study has been to characterize adult human somatic periodontium-derived stem cells (PDSCS) isolated from human periodontium and to follow their differentiation after cell culture. PDSCS were isolated from human periodontal tissue and cultured as spheres in serum-free medium. After 10 days the primary spheres were dissociated and the secondary spheres sub-cultured for another 1-2 weeks. Cells from different time points were analyzed, and immunohistochemical and electron microscopic investigations carried out. Histological analysis showed differentiation of spheres deriving from the PDSCS with central production of extracellular matrix beginning 3 days after sub-culturing. Isolated PDSCS developed pseudopodia which contained actin. Tubulin was found in the central portion of the cells. Pseudopodia between different cells anastomosed, indicating intercellular transport. Immunostaining for osteopontin demonstrated a positive reaction in primary spheres and within extracellular matrix vesicles after sub-culturing. In cell culture under serum-free conditions human PDSCS form spheres which are capable of producing extracellular matrix. Further investigations have do be carried out to investigate the capability of these cells to differentiate into osteogenic progenitor cells.
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Células Madre Adultas/citología , Ligamento Periodontal/citología , Adulto , Células Madre Adultas/ultraestructura , Técnicas de Cultivo de Célula , Diferenciación Celular , Forma de la Célula , Extensiones de la Superficie Celular/ultraestructura , Células Cultivadas , Medio de Cultivo Libre de Suero , Matriz Extracelular/química , Matriz Extracelular/ultraestructura , Humanos , Osteopontina/análisis , Seudópodos/ultraestructuraRESUMEN
The plasticity of dental pulp stem cells (DPSCs) has been demonstrated by several studies showing that they appear to self-maintain through several passages, giving rise to a variety of cells. The aim of the present study was to differentiate DPSCs to mature neuronal cells showing functional evidence of voltage gated ion channel activities in vitro. First, DPSC cultures were seeded on poly-l-lysine coated surfaces and pretreated for 48h with a medium containing basic fibroblast growth factor and the demethylating agent 5-azacytidine. Then neural induction was performed by the simultaneous activation of protein kinase C and the cyclic adenosine monophosphate pathway. Finally, maturation of the induced cells was achieved by continuous treatment with neurotrophin-3, dibutyryl cyclic AMP, and other supplementary components. Non-induced DPSCs already expressed vimentin, nestin, N-tubulin, neurogenin-2 and neurofilament-M. The inductive treatment resulted in decreased vimentin, nestin, N-tubulin and increased neurogenin-2, neuron-specific enolase, neurofilament-M and glial fibrillary acidic protein expression. By the end of the maturation period, all investigated genes were expressed at higher levels than in undifferentiated controls except vimentin and nestin. Patch clamp analysis revealed the functional activity of both voltage-dependent sodium and potassium channels in the differentiated cells. Our results demonstrate that although most surviving cells show neuronal morphology and express neuronal markers, there is a functional heterogeneity among the differentiated cells obtained by the in vitro differentiation protocol described herein. Nevertheless, this study clearly indicates that the dental pulp contains a cell population that is capable of neural commitment by our three step neuroinductive protocol.