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Use of molecular beacons to image effects of titanium surface microstructure on β1 integrin expression in live osteoblast-like cells

Paper ID Volume ID Publish Year Pages File Format Full-Text
7726 562 2010 8 PDF Available
Title
Use of molecular beacons to image effects of titanium surface microstructure on β1 integrin expression in live osteoblast-like cells
Abstract

This study used molecular beacon technology to examine substrate-dependent changes in integrin subunit expression in living cells. Molecular beacons are oligonucleotide probes that can be delivered into live cells to allow for real-time imaging of mRNA. They have a stem-loop hairpin structure with a fluorophore-quencher pair, which opens when bound to the target mRNA sequence, resulting in a fluorescent signal upon excitation. A novel molecular beacon that is specific to the β1 integrin subunit mRNA was developed and used to image osteoblast-like MG63 cells in vitro on both glass and titanium surfaces of varying roughness. Specificity was verified by comparing the molecular beacon signal intensities to real-time PCR results in both wild-type cells and cells with shRNA knockdown of β1 integrin mRNA. The molecular beacon was able to detect changes due to both surface microtopography and silencing of the mRNA target. The results showed that effects of the substrate on β1 mRNA noted previously in confluent cultures were evident in pre-confluent cells as well, supporting the hypothesis that β1 integrin pairs are important in proliferation as well as differentiation of osteoblasts. This technique overcomes the limitations of traditional gene assays (PCR, immunofluorescence) by allowing for the real-time measurement and tracking of specific mRNAs in individual live cells prior to confluence.

Keywords
Gene expression; Molecular imaging; Osteoblast; Titanium; Integrin
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Use of molecular beacons to image effects of titanium surface microstructure on β1 integrin expression in live osteoblast-like cells
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biomaterials - Volume 31, Issue 30, October 2010, Pages 7640–7647
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us