Speaker
Description
"Introduction
Mechanotransduction is a key process in many developmental, physiological and pathological processes in bone. Mechanosensitive ion channels such as Piezo1 and TREK1 are the bonafide mechanotransducers that are critical for various biological processes, plays a critical role in bone regeneration1, 2. We already reported that the Magnetic ion channel activation (MICA) technology could apply mechanical force directly to mechanosensitive ion channels on the cell surface targeted with antibody functionalised magnetic nanoparticles for stimulating mechanotransduction and downstream processes3. Recently there is an augmented interest has been aroused to exploit Graphene Oxide (GO) and its derivatives for various biomedical applications. The abundant oxygen-containing groups in GO provides an excellent platform for further modifications using functionalised antibodies to facilitate targeted binding4. Therefore, we have developed a potential nanoplatform using magnetic nanoparticles and GO (GOMNPs) for enhanced osteogenic differentiation through the MICA based non-invasive remote activation. GOMNPs targeting the mechanosensitive Piezo1 and TREK1 ion channels were developed and their osteogenic potential under MICA application is studied with osteoblast-like MG-63 cells.
Experimental details
Characterisation of synthesised GOMNPs was done using TEM, XRD, Raman spectroscopy and VSM. All the cell culture experiments were done on MG-63 human osteosarcoma cells using GOMNPs. Effect of MICA stimulation on osteogenic differentiation was studied with ALP activity, Alizarin red and PCR studies of GOMNPs functionalised with Piezo1 and TREK1 under MICA application for 1h daily for 7,14 and 21 days.
Results & Discussion
The biocompatible GOMNPs were prepared using a simple, versatile strategy. The size and morphology of the as-prepared magnetic nanoparticles and the GO were confirmed using TEM and the structural features of the GOMNPs were evaluated by X-ray diffraction XRD and Raman patterns. The XRD and Raman patterns indicating the peak positions and the relative intensities of MNPs, GO and the GOMNP nanocomposites are successfully obtained. The obtained results also suggested that the synthesised GOMNPs showed excellent biocompatibility with superparamagnetic behaviour. The enhanced ALP activity and Alizarin red staining data indicated the positive effect of MICA stimulation on MG63 cells towards osteogenic differentiation. MICA stimulation mediated osteogenic gene expression by quantitative real-time PCR also confirmed the differentiation of MG-63 cells at mRNA level and improved expression of a panel of osteogenic markers, Runx2, ALP and OCN were obtained upon 7th and 14th days under MICA application
Conclusion
The preliminary in vitro results demonstrated the ability of functionalised GOMNPs for osteogenic differentiation under MICA treatment. The studies also proved that the functionalised GOMNPs were able to successfully bind with the mechanosensitive TREK1 and Piezo1 ion channels and was able to enhance the osteogenesis through mechanotransduction via MICA.
References
1. Henstock, J. R.; Rotherham, M.; El Haj, A. J. J Tissue Eng 2018, 9.
2. Sun, W. J.; Chi, S. P.; Li, Y. H.; et al. Elife 2019, 8.
3. Hughes, S.; McBain, S.; Dobson, J.; El Haj, A. J. J R Soc Interface 2008, 5, (25), 855-863.
4. Sasikala, A. R. K.; Unnithan, A. R.; Thomas, R. G. et al. Adv Funct Mater 2018, 28, (8)."
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