Calcium phosphate-polymer hybrid nanoparticles for enhanced triple negative breast cancer treatment via co-delivery of paclitaxel and miR-221/222 inhibitors

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Calcium phosphate-polymer hybrid nanoparticles for enhanced triple negative breast cancer treatment via co-delivery of paclitaxel and miR-221/222 inhibitors Zilan Zhou, BS, Carly Kennell, MS, Joo-Youp Lee, PhD, Yuet-Kin Leung, PhD, Pheruza Tarapore, PhD Nanomedicine: Nanotechnology, Biology and Medicine Volume 13, Issue 2, Pages 403-410 (February 2017) DOI: 10.1016/j.nano.2016.07.016 Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 1 (A) Synthesis route of co-delivery nanoparticles (NP(miRi/pac)); (B) TEM images of (a) lipid-coated calcium phosphate miRi complex (lipid/CaP/miRi) and (b) co-delivery nanoparticle NP(miRi/pac); and (C) particle size distributions of (a) lipid/CaP/miRi complex and (b) NP(miRi/pac). Arrows indicate the lipid/CaP/miRi complex inside the NP(pac/miRi). Nanomedicine: Nanotechnology, Biology and Medicine 2017 13, 403-410DOI: (10.1016/j.nano.2016.07.016) Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 2 (A) encapsulation efficiencies of miRi-221/222 (miRi), scrambled miRi (miRiNC) and paclitaxel (pac) by nanoparticles. Data represent mean±standard deviation, n=4; and (B) in vitro release kinetics of miRi (black) and pac (red) from co-delivery nanoparticle NP (miRi/pac) at pH 7 (■) and pH5 (●), the amounts of miRi and paclitaxel remained inside NP(pac/miRi) were measured at 20h, 42h and 66h using fluorescence plate reader and HPLC respectively. Data represent mean±standard deviation, n=3. Nanomedicine: Nanotechnology, Biology and Medicine 2017 13, 403-410DOI: (10.1016/j.nano.2016.07.016) Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 3 Intracellular distribution of FAM-labeled miRi (green) delivered by NP(miRi) over MDA-MB-231. Nuclei were stained by 4′,6-diamidino-2-phenylindole (DAPI) (blue); endolysosomes were stained by LysoTracker (red). The cells were incubated with NP(miRi) at a miRi concentration of 20 pmol/ml for 5min, 30min, 2h, and 4h at 37°C. Arrows indicate miRi in the cytoplasm. Nanomedicine: Nanotechnology, Biology and Medicine 2017 13, 403-410DOI: (10.1016/j.nano.2016.07.016) Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 4 MicroRNA (miR) expression levels of (A) miR-221 and (B) miR-222 after 3-day treatment with NP(miRi) and NP(miRiNC) and mRNA expression levels of (C) p27 and (D) TIMP3 after 3-day treatment with NP(miRi) and NP(miRiNC).miRiNC with scrambled sequence serves as a negative control. Untreated (UT) cells were used as a control. Data represent mean±standard deviation, n=3. *P<0.05 vs. UT. Nanomedicine: Nanotechnology, Biology and Medicine 2017 13, 403-410DOI: (10.1016/j.nano.2016.07.016) Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 5 Cell viability for MDA-MB-231 after 3-day treatment with free drug (pac), NP(pac), NP(pac/miRiNC) and NP(pac/miRi). miRiNC with scrambled sequence serves as a negative control.For NP(pac/miRi) and NP(pac/miRiNC), the pac to miRi ratio is 1μg pac to 20 pmol miRi. Data represent mean±standard deviation, n=3. *P<0.05 vs. pac. Nanomedicine: Nanotechnology, Biology and Medicine 2017 13, 403-410DOI: (10.1016/j.nano.2016.07.016) Copyright © 2016 Elsevier Inc. Terms and Conditions

Nanomedicine: Nanotechnology, Biology and Medicine 2017 13, 403-410DOI: (10.1016/j.nano.2016.07.016) Copyright © 2016 Elsevier Inc. Terms and Conditions