KMID : 0869020100130010035
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Journal of Korean Orthopaedic Research Society 2010 Volume.13 No. 1 p.35 ~ p.42
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Affection of Wnt/¥â-catenin in Titanium Particles Challenged Osteoblasts
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Nam Ju-Suk
Nidi Sinha Sharma Ashish R Lee Jin-Koo Kwon Sun-Chang Chang Jun-Dong Lee Sang-Soo
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Abstract
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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/¥â-catenin signaling pathway in impaired osteogenesis, we analyzed the stability of ¥â-catenin and the transcriptional changes of regulators for Wnt/¥â-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 ¥â-catenin and GSK3¥âwas 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 ¥â-catenin and phospho-GSK3¥â were detected. Using immunofluorescence stain, it was confirmed that Ti-particles suppressed nucleus staining of ¥â-atenin 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/¥â-catenin signaling pathway in osteoblast through the transcriptional changes of membrane receptors and extracellular inhibitors for Wnt.
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KEYWORD
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Wnt/ ¥â-catenin, Osteoblast, Titanium particle
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