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


RESULTS
We 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.


CONCLUSIONS
Although 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.
Department of Bioengineering, University of Louisville, 505 S. Hancock, CTRB 623, Louisville, KY, 40208, USA.
Lee B
LB
Sims
Department of Bioengineering, University of Louisville, 505 S. Hancock, CTRB 623, Louisville, KY, 40208, USA.
Louis T
LT
Curtis
Department of Bioengineering, University of Louisville, 505 S. Hancock, CTRB 623, Louisville, KY, 40208, USA.|James Graham Brown Cancer Center, University of Louisville, Louisville, KY, USA.|Department of Pharmacology and Toxicology, University of Louisville, Louisville, KY, USA.
Hermann B
HB
Frieboes
Department of Bioengineering, University of Louisville, 505 S. Hancock, CTRB 623, Louisville, KY, 40208, USA. jill.steinbach@louisville.edu.|Department of Pharmacology and Toxicology, University of Louisville, Louisville, KY, USA. jill.steinbach@louisville.edu.|Department of Microbiology and Immunology, University of Louisville, Louisville, KY, USA. jill.steinbach@louisville.edu.|Center for Predictive Medicine, University of Louisville, Louisville, KY, USA. jill.steinbach@louisville.edu.
Jill M
JM
Steinbach-Rankins
D002460:Cell Line; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004337:Drug Carriers / Q000008:administration & dosage; D016503:Drug Delivery Systems / Q000379:methods; D005260:Female; D006367:HeLa Cells; D006801:Humans; D019344:Lactic Acid / Q000008:administration & dosage*; D053758:Nanoparticles / Q000008:administration & dosage*; D010316:Particle Size; D011092:Polyethylene Glycols / Q000008:administration & dosage; D011100:Polyglycolic Acid / Q000008:administration & dosage*; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D002583:Uterine Cervical Neoplasms / Q000188:drug therapy*
D004337:Drug Carriers; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D011092:Polyethylene Glycols
3D cell culture; Cell penetrating peptide; Cervical cancer; MPG; Nanoparticles; PEG; PLGA; Tumor spheroids; Vaginal epithelium; Vimentin