Abstract
Here we describe a method for imaging the position of nanoparticles within a 3D tissue-engineered model using confocal laser scanning microscopy (CLSM). The ability to track diffusion of nanoparticles in vitro is an important part of trans-dermal and trans-mucosal drug delivery development as well as for intra-epithelial drug delivery. Using 3D tissue-engineered models enables us to image diffusion in vitro in a physiologically relevant way; not possible in two-dimensional monolayer cultures (MacNeil, Nature 445:874–880, 2007; Hearnden et al., Pharmaceutical Res. 26(7):1718–1728, 2009). CLSM enables imaging of viable in vitro models in three dimensions with good spatial and axial resolution (Georgakoudi et al., Tissue Eng 14:1–20, 2008; Schenke-Layland et al., Adv. Drug Del. Rev. 58:878–896, 2006). Here we show that fluorescently labelled nanoparticles can be visualised, quantified, and their position within the cell can be determined using CLSM.
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References
Bhargava, S., Chapple, C. R., Bullock, A. J., Layton, C., and MacNeil, S. (2004) Tissue-engineered buccal mucosa for substitution urethroplasty. BJU Int. 93, 807–811.
Moharamzadeh, K., Brook, I. M., Van Noort, R., Scutt, A. M., and Thornhill, M. H. (2007) Tissue-engineered oral mucosa: a review of the scientific literature. J. Dent. Res. 86, 115–124.
Lomas, H., Massignani, M., Abdullah, K. A., Canton, I., Lo Presti, C., MacNeil, S., Du, J., Blanazs, A., Madsen, J., Armes, S. P., Lewis, A. L., and Battaglia, G. (2008) Non-cytotoxic polymer vesicles for rapid and efficient intracellular delivery. Faraday Discuss. 139, 143.
Lomas, H., Canton, I., MacNeil, S., Du, J., Armes, S. P. A., Ryan, A. J., Lewis, A. L., and Battaglia, G. (2007) Biomimetic pH sensitive polymersomes for efficient DNA encapsulation and delivery. Adv. Mater. 19, 4238–4243.
Hearnden, V., Lomas, H., MacNeil, S., Thornhill, M., Murdoch, C., Lewis, A. L., Madsen, J., Blanazs, A., Armes, S. P., and Battaglia, G. (2009) Diffusion studies of nanometer polymersomes across tissue engineered human oral mucosa. Pharm. Res. 26(7), 1718–1728.
Georgakoudi, I., Rice, W. L., Hronik-Tupaj, M., and Kaplan, D. L. (2008) Optical spectroscopy and imaging for the noninvasive evaluation of engineered tissues. Tissue Eng. 14, 1–20.
Muller, M. (2006) Introduction to confocal microscopy. 2nd ed., SPIE – The international society for optical engineering, USA.
Schenke-Layland, K., Riemann, I., Damour, O., Stock, U. A., and König, K. (2006) Two-photon microscopes and in vivo multiphoton tomographs – powerful diagnostic tools for tissue engineering and drug delivery. Adv. Drug Deliv. Rev. 58, 878–896.
Du, J., Tang, Y., Lewis, A. L., and Armes, S. P. (2005) pH-sensitive vesicles based on a biocompatible zwitterionic diblock copolymer. J. Am. Chem. Soc. 127, 17982–17983.
Zeiss, C. Chapter 4 Operating in Expert Mode. In Carl Zeiss LSM 510 Operating Manual, pp 280–285.
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Hearnden, V., MacNeil, S., Battaglia, G. (2011). Tracking Nanoparticles in Three-Dimensional Tissue-Engineered Models Using Confocal Laser Scanning Microscopy. In: Haycock, J. (eds) 3D Cell Culture. Methods in Molecular Biology, vol 695. Humana Press. https://doi.org/10.1007/978-1-60761-984-0_3
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DOI: https://doi.org/10.1007/978-1-60761-984-0_3
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