J Korean Orthop Res Soc.
2010 Jun;13(1):35-42.
Affection of Wnt/beta-catenin in Titanium Particles Challenged Osteoblasts
- Affiliations
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- 1Institute for Skeletal Aging & Orthopedic Surgery, Hallym University College of Medicin, Chuncheon, Korea. totalhip@hallym.ac.kr
- 2Institute for Natural Medicine, Hallym University College of Medicin, Chuncheon, Korea.
- 3Infectious Disease Medical Research Center, Hallym University College of Medicin, Chuncheon, Korea.
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.
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