Enhanced uptake and transport of PLGA-modified nanoparticles in cervical cancer

Abstract

BackgroundUncoordinated cellular proliferation and dysregulated angiogenesis in solid tumors are coupled with inadequate tissue, blood, and lymphatic vascularization. Consequently, tumors are often characterized by hypoxic regions with limited access to vascular-borne substances. In particular, systemically administered nanoparticles (NPs) targeting tumor cells and relying on vascular access to reach tumor tissue can suffer from limited therapeutic efficacy due to inhomogeneous intra-tumor distribution and insufficient cellular internalization of NPs. To circumvent these challenges, NP surfaces can be modified to facilitate tumor interstitial transport and cellular uptake.ResultsWe create poly(lactic-co-glycolic) acid NPs modified with MPG, polyethylene glycol (PEG), MPG/PEG, and Vimentin (VIM), and evaluate their cellular uptake in 2D (monolayer) cell culture of human cervical carcinoma (HeLa). We compare NP performance by evaluating uptake by non-cancerous vaginal (VK2) cells. We further assess NP interstitial transport in hypo-vascularized lesions by evaluating the effect of the various modifications on NP penetration in 3D cell culture of the HeLa cells. Results show that after 24 h incubation with HeLa cells in monolayer, MPG, MPG/PEG, PEG, and VIM NPs were internalized at 66×, 24×, 30×, and 15× that of unmodified NPs, respectively. In contrast, incubation with VK2 cells in monolayer showed that MPG , MPG/PEG , PEG , and VIM NPs internalized at 6.3×, 4.3×, 12.4×, and 3.0× that of unmodified NPs, respectively. Uptake was significantly enhanced in tumorigenic vs. normal cells, with internalization of MPG NPs by HeLa cells being twice that of PEG NPs by VK2 cells. After 24 h incubation in HeLa 3D cell culture, MPG and MPG/PEGNPs were internalized 2× and 3× compared to PEG and VIM NPs, respectively. Whereas MPG NPs were internalized mostly in the cell culture periphery (1.2×, 1.4×, and 2.7× that of PEG, MPG/PEG, and VIM NPs, respectively), PEG NPs at 250 μm penetrated 2× farther into the tissue culture than MPG NPs. For all NP types, cellular internalization was severely hindered in 3D compared to monolayer.ConclusionsAlthough MPG surface modification enhances internalization and uptake in hypo-vascularized cervical tissue culture, coating with PEG reduces this internalization while enhancing penetration. A delivery strategy combining NPs with either modification may balance cellular internalization vs. tissue penetration in hypo-vascularized cervical cancer lesions.Electronic supplementary materialThe online version of this article (doi:10.1186/s12951-016-0185-x) contains supplementary material, which is available to authorized users.

Peptide-modified nanoparticles inhibit formation of <em>Porphyromonas gingivalis</em> biofilms with <em>Streptococcus gordonii</em>

“…18,21 Here, we produced spherical NPs with zeta potential values for unmodified (-25 mV) and BAR-modified NPs (-7 mV) that are consistent with other avidin-NPs studies. 19,20 Since unmodified NPs have a negative surface charge, this difference can be attributed to the successful conjugation of positively charged avidin and BAR peptide to the NPs surface. In addition to confirming successful conjugation, the more positive surface charge exhibited by avidin-NPs and BARmodified NPs may also facilitate electrostatic interactions of NPs with negatively charged moieties on the P. gingivalis cell surface.…”

“…[18][19][20] Briefly, 10 mg of avidin was dissolved in 1.2 mL of 2% (w/v) sodium deoxycholate (NaDC) in PBS and warmed to 37°C. A 1 mg/mL palmitic acid-Nhydroxysuccinimide ester (PA-NHS; Sigma-Aldrich, St Louis, MO, USA) solution was prepared in 2% (w/v) NaDC and sonicated until well-mixed.…”

“…C6-containing NPs were synthesized using an oil-in-water (o/w) single-emulsion technique. [19][20][21] Briefly, C6 was encapsulated in 50-200 mg PLGA carboxylterminated polymer (0.55-0.75 dL/g; LACTEL ® ; DURECT Corporation, Cupertino, CA, USA). C6 was dissolved in 200 µL dichloromethane (DCM) overnight at a concentration of 15 µg/mg PLGA.…”

“…19,20 In this study, we synthesized unmodified, avidin-modified, and BAR-modified NPs and determined the overall NPs size distribution for each sample (Figure 1). The size distribution of unhydrated modified NPs trended smaller than unmodified NPs, but the average diameters of unmodified, avidin-modified, and BARmodified NPs calculated from SEM images (134±28 nm, 107±25 nm, and 99±29 nm, respectively; Table 1) were not statistically significant (P.0.05).…”

IL‑8 is upregulated in cervical cancer tissues and is associated with the proliferation and migration of HeLa cervical cancer cells

“…The human cervical carcinoma immortalized cell line HeLa was obtained from the American Type Culture Collection (Manassas, VA, USA). Cells were maintained in RPMI-�640 medium (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) and supplemented with �0% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc.) and penicillin-streptomycin (�00 units/ml) under standard culture conditions (12).…”