ICRS-PAT 2021

Enhancing inflammation targeting using tunable leukocyte-based biomimetic nanoparticles

Assaf Zinger 1 Caroline Cvetkovic Manuela Sushnitha 1 Tomoyuki Naoi 1 Gherardo Baudo 1 Enrica De Rosa 1 Jenny Chang 2 Morgan Anderson Arya Shetty Ennio Tasciotti 1 Francesca Taraballi 1 Nupur Basu Jennifer Covello Ennio Tasciotti Moran Amit
1Center for Musculoskeletal Regeneration; Orthopedics and Sports Medicine, Houston Methodist Academic Institute; Houston Methodist Hospital, USA
2Houston Methodist Cancer Center, Houston Methodist Research Institute, USA

Biomimetic nanoparticles aim to effectively emulate the behavior of either cells or exosomes.1 Leukocyte-based biomimetic nanoparticles, for instance, incorporate cell membrane proteins to transfer the natural tropism of leukocytes to the final delivery platform. However, tuning the protein integration can affect the in vivo behavior of these nanoparticles and alter their efficacy. Here we show that, while increasing the protein:lipid ratio to a maximum of 1:20 (w/w) maintained the nanoparticle’s structural properties, increasing protein content resulted in improved targeting of inflamed endothelium in two different animal models. Our combined use of a microfluidic, bottom-up approach and tuning of a key synthesis parameter enabled the synthesis of reproducible, enhanced biomimetic nanoparticles that have the potential to improve the treatment of inflammatory-based conditions through targeted nanodelivery (Fig. 1).2

References:

1. Zinger, A., Brozovich, A., Pasto, A., Sushnitha, M., Martinez, J.O., Evangelopoulos, M., Boada, C., Tasciotti, E. and Taraballi, F., 2020. Bioinspired extracellular vesicles: lessons learned from nature for biomedicine and bioengineering. Nanomaterials, 10(11), p.2172.

2. Zinger, A., Sushnitha, M., Naoi, T., Baudo, G., De Rosa, E., Chang, J., Tasciotti, E. and Taraballi, F., 2021. Enhancing inflammation targeting using tunable leukocyte-based biomimetic nanoparticles. ACS nano, 15(4), pp.6326-6339.

Figure 1. Schematic of the different protein:lipid ratios (P:L) biomimetic NP microfluidic synthesis, characterization, and in vitro and in vivo experiments. (1) Leukocytes (J774 cell line) were cultivated in vitro and used to extract (2) membrane proteins for the synthesis of (3) different P:L NPs using a microfluidic approach by increasing the membrane protein concentration in the aqueous phase. For Lipo, no membrane proteins were added, while we added 0.058 mg/mL of protein extract for Leuko1:100, 0.145 mg/mL for Leuko1:40, and 0.29 mg/mL for Leuko 1:20. The physical, chemical, and biological properties of the NP were characterized (4) and then tested in vitro using inflamed endothelial cells (5). Finally, the enhanced targeting was evaluated in vivo using LLI and TNBC models. Figure 1 was created using Biorender.com.









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