RESUMEN
It has been previously reported that caveolin-1 (Cav-1) knockout mice exhibit increased bone size and stiffness. However, the expression and role of Cav-1 on periodontal tissue is poorly understood. The aim of this study was to investigate the immunohistochemical expression of Cav-1 in the mouse periodontium and explore the role of Cav-1 on osteoblastic and cementoblastic differentiation in human periodontal ligament cells (hPDLCs), cementoblasts, and osteoblasts. To reveal the molecular mechanisms of Cav-1 activity, associated signaling pathways were also examined. Immunolocalization of Cav-1 was studied in mice periodontal tissue. Differentiation was evaluated by ALP activity, alizarin red S staining, and RT-PCR for marker genes. Signal transduction was analyzed using Western blotting and confocal microscopy. Cav-1 expression was observed in hPDLCs, cementoblasts, and osteoblasts of the periodontium both in vivo and in vitro. Inhibition of Cav-1 expression by methyl-ß-cyclodextrin (MßCD) and knockdown of Cav-1 by siRNA promoted osteoblastic and cementoblastic differentiation by increasing ALP activity, calcium nodule formation, and mRNA expression of differentiation markers in hPDLCs, cementoblasts, and osteoblasts. Osteogenic medium-induced BMP-2 and BMP-7 expression, and phosphorylation of Smad1/5/8 were enhanced by MßCD and siRNA knockdown of Cav-1, which was reversed by BMP inhibitor noggin. MßCD and Cav-1 siRNA knockdown increased OM-induced AMPK, Akt, GSK3ß, and CREB phosphorylation, which were reversed by Ara-A, a specific AMPK inhibitor. Moreover, OM-induced activation of p38, ERK, JNK, and NF-κB was enhanced by Cav-1 inhibition. This study demonstrates, for the first time, that Cav-1 is expressed in developing periodontal tissue and in vitro in periodontal-related cells. Cav-1 inhibition positively regulates osteoblastic differentiation in hPDLCs, cementoblasts, and osteoblasts via BMP, AMPK, MAPK, and NF-κB pathway. Thus, Cav-1 inhibition may be a novel molecular target for therapeutic approaches in periodontitis or osteolytic disease.
Asunto(s)
Caveolina 1/biosíntesis , Cemento Dental/citología , Osteoblastos/citología , Periodoncio/citología , Periodoncio/metabolismo , Animales , Western Blotting , Diferenciación Celular/fisiología , Células Cultivadas , Femenino , Humanos , Inmunohistoquímica , Técnicas In Vitro , Ratones , Ratones Endogámicos ICR , Ligamento Periodontal/citología , Ligamento Periodontal/crecimiento & desarrollo , Periodoncio/crecimiento & desarrollo , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transducción de Señal/fisiologíaRESUMEN
Baicalin (BC) has been used for cancer therapy due to its multiple effects as an anti-cancer drug. However, the effective delivery of this molecule to targeted cells is difficult. Gold nanoparticles (AuNPs) conjugated with thiolated beta cyclodextrin (AuNP-S-ß-CD) were used as a delivery vector in this study. Cell viability tests were evaluated by cell counting kit-8 (CCK) and live/dead cell assay. To demonstrate the proliferation inhibition effect on Michigan Cancer Foundation-7 (MCF-7) cells by BC, we analyzed using Hoechst 33342 staining assay and gel electrophoresis. The S-ß-CD conjugated AuNPs were characterized by transmission electron microscopy (TEM), 1H nuclear magnetic resonance ((1)H NMR), and ultraviolet visible (UV-vis) spectroscopy. AuNP-S-ß-CD with approximately 40 µM of BC loaded by inclusion complex showed an inhibition effect on MCF-7 cells by inducing apoptosis. Apoptosis test results were evaluated by analyzing the expression of typical apoptic markers such as cleaved caspase-3, full length caspase-3, and apaf-1 in western blot assay. These results demonstrated that AuNP-S-ß-CD-BC inhibited the proliferation of cancerous MCF-7 cells by inducing apoptosis. These findings suggested that AuNP-S-ß-CD-BC could be a promising agent for chemotherapeutic usage.
RESUMEN
In this study, we developed poly(É-caprolactone) (PCL) 3D scaffolds using a solid free form fabrication (SFF) technique. ß-cyclodextrin (ßCD) was grafted to hydroxyapatite (HAp) and this ßCD grafted HAp was coated onto the PCL scaffold surface, followed by drug loading through an inclusion complex interaction between the ßCD and adamantane (AD) or between ßCD and simvastatin (SIM). The scaffold structure was characterized by scanning electron microscopy (SEM). The release profile of simvastatin in the ß-CD grafted HAp was also evaluated. Osteogenic differentiation of adipose-derived stromal cells (ADSCs) was examined using an alkaline phosphatase activity (ALP) assay. The results suggest that drug loaded PCL-HAp 3-D scaffolds enhances osteogenic differentiation of ADSCs.
RESUMEN
PURPOSE: This study was performed to investigate the incidence and configuration of the bifid mandibular canal in a Korean population by using cone-beam computed tomography (CBCT) imaging. MATERIALS AND METHODS: CBCT images of 1933 patients (884 male and 1049 female) were evaluated using PSR-9000N and Alphard-Vega 3030 Dental CT units (Asahi Roentgen Ind. Co., Ltd, Kyoto, Japan). Image analysis was performed by using OnDemand3D software (CyberMed Inc., Seoul, Korea). The bifid mandibular canal was identified and classified into four types, namely, the forward canal, buccolingual canal, dental canal, and retromolar canal. Statistical analysis was performed by using the chi-squared test and one-way analysis of variance (ANOVA). RESULTS: Bifid mandibular canals were observed in 198 (10.2%) of 1933 patients. The most frequently observed type of bifid mandibular canal was the retromolar canal (n=104, rate: 52.5%) without any significant difference among the incidence of each age and gender. The mean diameter of the accessory canal was 1.27 mm (range: 0.27-3.29 mm) without any significant difference among the mean diameter of each type of the bifid mandibular canal. The mean length of the bifid mandibular canals was 14.97mm(range: 2.17-38.8 mm) with only a significant difference between the dental canal and the other types. CONCLUSION: The bifid mandibular canal is not uncommon in Koreans and has a prevalence of 10.2% as indicated in the present study. It is suggested that a CBCT examination be recommended for detecting a bifid canal.