Affection of Wnt/beta-catenin in Titanium Particles Challenged Osteoblasts.
- Author:
Ju Suk NAM
1
;
Sinha NIDI
;
Ashish R SHARMA
;
Jin Koo LEE
;
Sun Chang KWON
;
Jun Dong CHANG
;
Sang Soo LEE
Author Information
1. Institute for Skeletal Aging & Orthopedic Surgery, Hallym University College of Medicin, Chuncheon, Korea. totalhip@hallym.ac.kr
- Publication Type:Original Article
- Keywords:
Wnt/beta-catenin;
Osteoblast;
Titanium particle
- MeSH:
beta Catenin;
Blotting, Western;
Culture Media, Conditioned;
Fluorescent Antibody Technique;
Glycogen Synthase Kinase 3;
Membranes;
Microscopy;
Osteoblasts;
Osteogenesis;
Osteolysis;
RNA, Messenger;
Titanium
- From:Journal of Korean Orthopaedic Research Society
2010;13(1):35-42
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
PURPOSE: The intracellular mechanisms that lead to periprosthetic osteolysis including impaired bone forming activity of osteoblast remain incompletely characterized. To determine the possibility that Ti-particles play a role to regulate Wnt/beta-catenin signaling pathway in impaired osteogenesis, we analyzed the stability of beta-catenin and the transcriptional changes of regulators for Wnt/beta-catenin signaling pathway in MC3T3-E1 osteoblast cells. MATERIALS AND METHODS: Ti-particles were prepared by sterilizing and counted on the microscopy. Transcriptional changes of OPG, RANKL, LRP5, LRP6, DKK1 and sFRP2 were determined by real-time RTPCR. Protein level of beta-catenin and GSK3beta was detected using Western blotting and immunofluorescence staining. RESULTS: After 4 hours of treatment of Ti-particles, OPG/RANKL mRNA ratio was significantly decreased. And also, decreased protein levels of beta-catenin and phospho-GSK3beta were detected. Using immunofluorescence stain, it was confirmed that Ti-particles suppressed nucleus staining of beta-catenin induced by Wnt3a conditioned medium. The results of real-time RT-PCR showed reduced level of LRP5 and LRP6 transcripts, and induced level of DKK1 and sFRP2 transcripts by challenging of Ti-particles CONCLUSION: Our report suggests that Ti-particles may play a crucial role in the regulation of Wnt/beta-catenin signaling pathway in osteoblast through the transcriptional changes of membrane receptors and extracellular inhibitors for Wnt.
