International Journal of Nanomedicine (Jul 2018)
Bioengineering the spider silk sequence to modify its affinity for drugs
Abstract
Kamil Kucharczyk,1,2 Marek Weiss,3 Katarzyna Jastrzebska,1,2 Magdalena Luczak,4,5 Arkadiusz Ptak,3 Maciej Kozak,6,7 Andrzej Mackiewicz,1,2 Hanna Dams-Kozlowska1,2 1Department of Medical Biotechnology, Poznan University of Medical Sciences, Poznan, Poland; 2Department of Diagnostics and Cancer Immunology, Greater Poland Cancer Centre, Poznan, Poland; 3Division of Computational Physics and Nanomechanics, Institute of Physics, Faculty of Technical Physics, Poznan University of Technology, Poznan, Poland; 4Department of Biomedical Proteomics, Institute of Bioorganic Chemistry, Polish Academy of Sciences, Poznan, Poland; 5Department of Organic Chemistry, Institute of Chemical Technology and Engineering, Poznan University of Technology, Poznan, Poland; 6Department of Macromolecular Physics, Adam Mickiewicz University, Poznan, Poland; 7Joint Laboratory for SAXS Studies, Adam Mickiewicz University, Poznan, Poland Background: Silk is a biocompatible and biodegradable material, able to self-assemble into different morphological structures. Silk structures may be used for many biomedical applications, including carriers for drug delivery. The authors designed a new bioengineered spider silk protein, EMS2, and examined its property as a carrier of chemotherapeutics.Materials and methods: To obtain EMS protein, the MS2 silk monomer (that was based on the MaSp2 spidroin of Nephila clavipes) was modified by the addition of a glutamic acid residue. Both bioengineered silks were produced in an Escherichia coli expression system and purified by thermal method. The silk spheres were produced by mixing with potassium phosphate buffer. The physical properties of the particles were characterized using scanning electron microscopy, atomic force microscopy, Fourier-transform infrared spectroscopy, and zeta potential measurements. The MTT assay was used to examine the cytotoxicity of spheres. The loading and release profiles of drugs were studied spectrophotometrically.Results: The bioengineered silk variant, EMS2, was constructed, produced, and purified. The EMS2 silk retained the self-assembly property and formed spheres. The spheres made of EMS2 and MS2 silks were not cytotoxic and had a similar secondary structure content but differed in morphology and zeta potential values; EMS2 particles were more negatively charged than MS2 particles. Independently of the loading method (pre- or post-loading), the loading of drugs into EMS2 spheres was more efficient than the loading into MS2 spheres. The advantageous loading efficiency and release rate made EMS2 spheres a good choice to deliver neutral etoposide (ETP). Despite the high loading efficiency of positively charged mitoxantrone (MTX) into EMS2 particles, the fast release rate made EMS2 unsuitable for the delivery of this drug. A faster release rate from EMS2 particles compared to MS2 particles was observed for positively charged doxorubicin (DOX).Conclusion: By modifying its sequence, silk affinity for drugs can be controlled. Keywords: silk, bioengineering, spheres, drug delivery, chemotherapeutics, cancer therapy
