Jeong B.-C.,Dental Science Research Institute |
Kim H.-J.,Dental Science Research Institute |
Bae I.-H.,Dental Science Research Institute |
Lee K.-N.,Dental Science Research Institute |
And 8 more authors.
Introduction: Angiogenesis is closely associated with bone formation, especially endochondral ossification. Angiopoietin 1 (Ang1) is a specific growth factor functioning to generate a stable and matured vasculature through the Tie2 receptor/PI3K/AKT pathway. Recently cartilage oligomeric matrix protein (COMP)-Ang1, an Ang1 variant which is more potent than native Ang1 in phosphorylating Tie2 receptor and AKT, was developed. This study was designed to examine the effects of angiogenic COMP-Ang1 on BMP2-induced osteoblast differentiation and bone formation. Methods: Expression of endogenous Ang-1 and its binding receptor Tie 2 mRNA was examined in osteoblast-like cells and primary mouse calvarial cells by RT-PCR analysis, and was also monitored during osteoblast differentiation induced by BMP-2 and/or ascorbic acid and β-glycerophosphate. Effects of COMP-Ang-1 on osteoblast differentiation and mineralization were evaluated by alkaline phosphatase (ALP) activity and osteocalcin (OC) production, and Alizarin red stain. For a molecular mechanism, Western blot and OG2 and 6xOSE promoter assays were done. For in vivo evaluation, adenoviral (Ad) vectors containing COMP-Ang-1 or BMP-2 gene were administered into thigh muscle of mice, and after 2 weeks bone formation was analyzed by micro-computed tomography and histology. Angiogenic event of COMP-Ang1 was confirmed by immunofluorescence analysis with anti-CD31 antibody. Results: Expression of Tie2 receptor was significantly increased in the course of osteoblast differentiation. Treatment or overexpression of COMP-Ang1 enhanced BMP2-induced ALP activity, OC production, and mineral deposition in a dose-dependent manner. In addition, COMP-Ang1 synergistically increased OG2 and 6xOSE promoter activities of BMP2, and sustained p38, Smad and AKT phosphorylation of BMP2. Notably, in vivo intramuscular injection of COMP-Ang1 dose-dependently enhanced BMP2-induced ectopic bone formation with increases in CD31 reactivity. Conclusions: These results suggest that COMP-Ang1 synergistically enhanced osteoblast differentiation and bone formation through potentiating BMP2 signaling pathways and angiogenesis. Combination of BMP2 and COMP-Ang1 should be clinically useful for therapeutic application to fracture and destructive bone diseases. © 2009 Elsevier Inc. All rights reserved. Source
Yun K.-D.,Chonnam National University |
Yang Y.,University of Texas Health Science Center at Houston |
Lim H.-P.,Chonnam National University |
Oh G.-J.,Dental Science Research Institute |
And 5 more authors.
Materials Science and Engineering C
This study was to evaluate wettability, cell response, and osseointegration of nanotubular titanium (Ti) surface by anodic oxidation. Commercially pure Ti discs were treated by polishing, sandblasting, and anodizing. These surfaces were characterized by scanning electron microscopy and contact angle measurement. MC3T3-E1 osteoblast cell was used to evaluate cell response in vitro. The cell morphology, cell viability, and alkaline phosphatase (ALP) specific activity were assessed. The Ti implants of 2.0 mm diameter and 5.0 mm long treated by anodizing and sandblasting/anodizing were inserted into the tibia of rats. After 3 weeks, the histology of the Ti-bone interface was examined. SEM observations showed that the anodizing and sandblasting/anodizing created the nanotubular surface and graded nanotubular-micro-roughened surfaces, respectively. The anodizing and sandblasting/anodizing significantly improved the hydrophilicity of Ti. The significant greatest cell spreading and ALP specific activity were observed on the graded nanotubular-micro-roughened surfaces treated by sandblasting/anodizing. The in vivo study shows that newly formed bone was intimately in contact with the nanotubular surfaces without adverse immune response. This study has suggested that the graded nanotubular-micro-roughened surface of Ti treated with sandblasting/anodizing is very promising in implantology due to improved hydrophilicity, favorable cell response, and excellent osseointegration. © 2009 Elsevier B.V. All rights reserved. Source
Chung K.-J.,Dental Science Research Institute |
Cho E.-J.,Dental Science Research Institute |
Kim M.K.,Dental Science Research Institute |
Kim Y.R.,Dongshin University |
And 9 more authors.
Journal of Infectious Diseases
Infection with the human pathogen Vibrio vulnificus leads to the generation of reactive oxygen species (ROS) via NAD(P)H oxidase (Nox) in host cells. In the present study, we employed mutant V. vulnificus strains to identify an essential virulence factor responsible for this ROS generation. We found that repeats-in-toxin A1 (RtxA1) expressed by V. vulnificus acts via Noxl to induce significant ROS generation in the intestine epithelial cells, which ultimately results in cell death. Furthermore, RtxA1 modulates the small GTPase Rac2, which is known to play an important role in the activation of Nox. When mice were infected by the oral method, in contrast with the wild-type bacteria, an RtxA1-deficient V. vulnificus mutant was unable to induce ROS generation within the intestine and failed to cause death. These findings strongly suggest that RtxA1-induced Rac2 expression is a critical step underlying the pathogenicity of V. vulnificus. © 2009 by the Infectious Diseases Society of America. All rights reserved. Source