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BACKGROUND: Maxillary/mandibular bone marrow-derived mesenchymal stem cells (MBMSCs) exhibit a unique property of lower adipogenic potential than other bone marrow-derived MSCs. However, the molecular mechanisms regulating the adipogenesis of MBMSCs remain unclear. This study aimed to explore the roles of mitochondrial function and reactive oxygen species (ROS) in regulating the adipogenesis of MBMSCs. METHODS AND RESULTS: MBMSCs exhibited significantly lower lipid droplet formation than iliac BMSCs (IBMSCs). Moreover, the expression levels of CCAAT/enhancer-binding protein ß (C/EBPß), C/EBPδ, and early B cell factor 1 (Ebf-1), which are early adipogenic transcription factors, and those of peroxisome proliferator-activated receptor-γ (PPARγ) and C/EBPα, which are late adipogenic transcription factors, were downregulated in MBMSCs compared to those in IBMSCs. Adipogenic induction increased the mitochondrial membrane potential and mitochondrial biogenesis in MBMSCs and IBMSCs, with no significant difference between the two cell types; however, intracellular ROS production was significantly enhanced only in IBMSCs. Furthermore, NAD(P)H oxidase 4 (NOX4) expression was significantly lower in MBMSCs than in IBMSCs. Increased ROS production in MBMSCs by NOX4 overexpression or treatment with menadione promoted the expression of early adipogenic transcription factors but did not induce that of late adipogenic transcription factors or lipid droplet accumulation. CONCLUSIONS: These results suggest that ROS may be partially involved in the process of MBMSC adipogenic differentiation from undifferentiated cells to immature adipocytes. This study provides important insights into the tissue-specific properties of MBMSCs.
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Adipogénesis , Células Madre Mesenquimatosas , Humanos , Adipogénesis/fisiología , Especies Reactivas de Oxígeno/metabolismo , Médula Ósea/metabolismo , Diferenciación Celular , Células Madre Mesenquimatosas/metabolismo , PPAR gamma/genética , PPAR gamma/metabolismo , Células de la Médula Ósea , Células CultivadasRESUMEN
OBJECTIVES: The purpose of this study was to evaluate the effect of low-serum STK2 medium on the isolation and osteogenic differentiation of human maxillary/mandibular bone marrow stromal cells (MBMSCs). MATERIALS AND METHODS: Human MBMSCs were obtained from patients undergoing dental implant treatment. These cells were cultured in serum-free medium or STK2 medium containing 1 % fetal bovine serum (low-serum) or α-MEM containing 10 % fetal bovine serum (control). Proliferation on the culture surface, cell surface antigen expression, and mRNA levels of neural crest and osteogenic markers were examined. Alkaline phosphatase assay and Alizarin red staining were used to assess osteogenic differentiation potential. Immunoblotting analysis was performed to detect ERK phosphorylation. RESULTS: Low-serum and control MBMSCs were positive for CD73, CD90, and CD105 and negative for CD14, CD34, CD45, CD271, and HLA-DR. CD140a was absent in low-serum cells but present in control cells. Low-serum MBMSCs proliferated more than control MBMSCs. After induction of osteogenic differentiation, alkaline phosphatase activity and osteocalcin mRNA levels were higher in low-serum MBMSCs than in control cells, and Alizarin red staining was stronger in low-serum MBMSCs than in control cells. Low-serum culture promoted ERK phosphorylation. CONCLUSIONS: MBMSCs precultured in low-serum medium exhibited a greater cumulative cell number and a higher osteogenic differentiation capacity than those cultured in control medium. CLINICAL RELEVANCE: These findings indicate that low-serum STK2 culture might be useful to promote MBMSC proliferation and osteogenic differentiation. This method requires less autologous blood collection for cell expansion than conventional methods, thus reducing the burden on patients.
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Proliferación Celular/efectos de los fármacos , Medio de Cultivo Libre de Suero/farmacología , Células Madre Mesenquimatosas/citología , Osteogénesis/efectos de los fármacos , Antígenos de Superficie/análisis , Células Cultivadas , Depresión Química , Humanos , Immunoblotting , Fosforilación , ARN Mensajero/análisis , Estimulación QuímicaRESUMEN
OBJECTIVE: This study aims to investigate the underlying molecular mechanisms that regulate the adipogenic differentiation of maxillary/mandibular bone marrow-derived mesenchymal stem cells (MBMSCs). DESIGN: MBMSCs and iliac bone marrow-derived MSCs (IBMSCs) were compared for osteogenic, chondrogenic, and adipogenic differentiation. Cell surface antigen expression was examined using flow cytometry, and stem cell marker expression was assessed using real-time polymerase chain reaction (PCR). Various adipogenic regulatory factors' expression was evaluated using real-time PCR and western blotting. RESULTS: No significant differences in cell surface antigen profiles or stem cell marker expression in MBMSCs and IBMSCs were observed. MBMSCs and IBMSCs displayed similar osteogenic and chondrogenic potentials, whereas MBMSCs showed significantly lower adipogenic potentials than those shown by IBMSCs. Expression of CCAAT/enhancer binding protein ß (C/EBPß), C/EBPδ, early B-cell factor 1 (Ebf-1), and Krüppel-like factor 5 (KLF5), which are early adipogenic differentiation factors, was suppressed in MBMSCs compared to that in IBMSCs. Peroxisome proliferator-activated receptor-γ (PPARγ) and C/EBPα, which play important roles in the terminal differentiation of adipocytes, was lower in MBMSCs than that in IBMSCs. Furthermore, the level of zinc finger protein 423 (Zfp423), which is involved in the commitment of undifferentiated MSCs to the adipocyte lineage, was significantly lower in MBMSCs than that in IBMSCs. CONCLUSIONS: MBMSCs are negatively regulated in the commitment of undifferentiated MSCs to the adipocyte lineage (preadipocytes) as well as in the terminal differentiation of preadipocytes into mature adipocytes. These results may elucidate the site-specific characteristics of MBMSCs.
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Adipogénesis , Médula Ósea , Humanos , Médula Ósea/metabolismo , Diferenciación Celular/fisiología , Adipogénesis/fisiología , Adipocitos/metabolismo , Factores de Transcripción/metabolismo , Células Madre/metabolismo , PPAR gamma/metabolismoRESUMEN
Preservation of the alveolar ridge after tooth extraction is an essential component for ideal implant positioning. Furthermore, preservation of bone around the implant after implant placement is an essential component for implant treatment. We aimed to evaluate the efficacy of bone grafting materials in preserving the alveolar ridge after implant placement. Implants were placed in regenerated bone without grafting material or with beta-tricalcium phosphate, bovine bone substitute, or carbonate apatite transplantation. In all groups, the bone healed and the implants were successfully placed within the bone. No significant differences in insertion torque and implant stability quotient values were found. The amount of bone around the implant 5 weeks after implant placement was significantly reduced in the bovine bone substitute group; however, implants placed in regenerated bone achieved sufficient initial fixation and osseointegration.
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Aumento de la Cresta Alveolar , Sustitutos de Huesos , Implantes Dentales , Proceso Alveolar , Animales , Trasplante Óseo , Bovinos , Implantación Dental Endoósea , Alveolo Dental/cirugíaRESUMEN
JH8194 induces osteoblast differentiation, although it was originally designed to improve antifungal activity. This suggests that JH8194 is useful for implant treatment. Therefore, the aim of this study was to evaluate the osseointegration capacity of JH8194-modified titanium dental implant fixtures (JH8194-Fi). The implants were randomly implanted into the edentulous ridge of dog mandibles. Healing abutments were inserted immediately after implant placement. Three weeks later, peri-implant bone levels, the first bone-to-implant contact points, and trabecular bone formation surrounding the implants were assessed by histological and digital image analyses based on microcomputed tomography (microCT). The histological analysis revealed an enhancement of mature trabecular bone around the JH8194-Fi compared with untreated fixtures (control-Fi). Similarly, microCT combined with analysis by Zed View™ also showed increased trabecular bone formation surrounding the JH8194-Fi compared with the control-Fi (Student's t-test, P < 0.05). JH8194 may offer an alternative biological modification of titanium surfaces to enhance trabecular bone formation around dental implants, which may contribute to the transient acquirement of osseointegration and the long-term success of implant therapy.
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Péptidos Catiónicos Antimicrobianos/administración & dosificación , Huesos/fisiología , Implantes Dentales , Histatinas/administración & dosificación , Titanio/química , Animales , Huesos/patología , Materiales Biocompatibles Revestidos , Perros , Histatinas/química , Mandíbula/patología , Oseointegración , Osteoblastos/citología , Prótesis e Implantes , Propiedades de Superficie , Tomografía Computarizada por Rayos X , Microtomografía por Rayos XRESUMEN
INTRODUCTION: Maxillary/mandibular bone marrow stromal cells (MBMSCs) are a useful cell source for bone regeneration in the oral and maxillofacial region. To further ensure the clinical application of MBMSCs in bone regenerative therapy, it is important to determine the bone formation capacity of MBMSCs before transplantation. The aim of this study is to identify the molecular marker that determines the in vivo bone formation capacity of MBMSCs. METHODS: The cell growth, cell surface antigens, in vitro and in vivo bone formation capacity of MBMSCs were examined. The amount of chitinase-3-like protein 1 (CHI3L1) secreted into the conditioned medium was quantified. The effects of CHI3L1 on the cell growth and osteogenic differentiation potential of MBMSCs and on the cell growth and migration of vascular endothelial cells and fibroblasts were examined. RESULTS: The cell growth, and in vitro and in vivo bone formation capacity of the cells treated with different conditions were observed. MBMSCs that secreted a large amount of CHI3L1 into the conditioned medium tended to have low in vivo bone formation capacity, whereas MBMSCs that secreted a small amount of CHI3L1 had greater in vivo bone formation capacity. CHI3L1 promoted the migration of vascular endothelial cells, and the cell growth and migration of fibroblasts. CONCLUSION: Our study indicates that the in vitro osteogenic differentiation capacity of MBMSCs and the in vivo bone formation capacities of MBMSCs were not necessarily correlated. The transplantation of high CHI3L1 secretory MBMSCs may suppress bone formation by inducing fibrosis at the site. These results suggest that the CHI3L1 secretion levels from MBMSCs may be used as a predictable marker of bone formation capacity in vivo.
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The purpose of this study was to evaluate the effects of commercialized denture adhesives and mouth moisturizers using direct and indirect cell cultures for in vitro examinations with human fibroblast and epithelial cells. Denture adhesives (Faston, Poligrip Powder, New Poligrip Free, Tafugurippu Kurimu, Polident Adhesive, Tafugurippu Tomei) and mouth moisturizers (Concool Mouth Gel, Biotene Oral Balance Gel) were subjected to live and dead detection and pH level determination. The mouth moisturizers showed higher cytotoxicity effects comparing with control on every cell cultures and cells, and pH level did not show any significant differences. However, there was no correlation of type of denture adhesive or mouth moisturizer with cytotoxicity. We concluded that cytotoxicity affects human cells regardless of type of material, though some dependence was noted.
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Retención de Dentadura , Queratinocitos , Adhesivos , Cementos Dentales , Fibroblastos , Humanos , BocaRESUMEN
Most of the cases of dental implant surgery, especially the bone defect extensively, are essential for alveolar ridge augmentation. As known as cell therapy exerts valuable effects on bone regeneration, numerous reports using various cells from body to regenerate bone have been published, including clinical reports. Mesenchymal cells that have osteogenic activity and have potential to be harvested from intra oral site might be a candidate cells to regenerate alveolar bone, even dentists have not been harvested the cells outside of mouth. This paper presents a summary of somatic cells in edentulous tissues which could subserve alveolar bone regeneration. The candidate tissues that might have differentiation potential as mesenchymal cells for bone regeneration are alveolar bone chip, bone marrow from alveolar bone, periosteal tissue, and gingival tissue. Understanding their phenotype consecutively will provide a rational approach for alveolar ridge augmentation.