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Enhanced Specificity and Drug Delivery in Tumors by cRGD - Anchoring Thermosensitive Liposomes.

Dicheva BM, Ten Hagen TL, Seynhaeve AL, Amin M, Eggermont AM, Koning GA - Pharm. Res. (2015)

Bottom Line: Cytotoxic effect of TSL and RGD-TSL was studied on B16Bl6 melanoma, B16F10 melanoma and HUVEC.High resolution intravital microscopy demonstrated specific accumulation of RGD-TSL to the tumor vasculature.Moreover, application of hyperthermia resulted in massive drug release from RGD-TSL.

View Article: PubMed Central - PubMed

Affiliation: Laboratory Experimental Surgical Oncology, Section Surgical Oncology Department of Surgery, Erasmus MC Cancer Center, Rotterdam, The Netherlands. b.dicheva@erasmusmc.nl.

ABSTRACT

Purpose: To develop RGD-targeted thermosensitive liposomes with increased tumor retention, improving drug release efficiency upon mild hyperthermia (HT) in both tumor and angiogenic endothelial cells.

Methods: Standard termosensitive liposomes (TSL) and TSL containing a cyclic Arg-Gly-Asp (cRGD) pentapeptide with the sequence Arg-Cys-D-Phe-Asp-Gly (RGDf[N-Met]C) were synthetized, loaded with Dox and characterized. Temperature- and time-dependent drug release profiles were assessed by fluorometry. Intracellular Dox delivery was studied by flow cytometry and confocal microscopy. Cytotoxic effect of TSL and RGD-TSL was studied on B16Bl6 melanoma, B16F10 melanoma and HUVEC. Intravital microscopy was performed on B16Bl6 tumors implanted in dorsal-skin fold window-bearing mice. Pharmacokinetic and biodistribution of Dox-TSL and Dox-RGD-TSL were followed in B16Bl6 tumor bearing mice upon normothermia or initial hyperthermia conditions.

Results: DLS and cryo-TEM revealed particle homogeneity and size of around 85 nm. Doxorubicin loading efficiency was >95%as assessed by spectrofluorometry. Flow cytometry and confocal microscopy showed a specific uptake of RGD-TSL by melanoma and endothelial cells when compared to TSL and an increased doxorubicin delivery. High resolution intravital microscopy demonstrated specific accumulation of RGD-TSL to the tumor vasculature. Moreover, application of hyperthermia resulted in massive drug release from RGD-TSL. Biodistribution studies showed that initial hyperthermia increases Dox uptake in tumors from TSL and RGD-TSL.

Conclusion: RGD-TSL have potency to increase drug efficacy due to higher uptake by tumor and angiogenic endothelial cells in combination with heat-triggered drug release.

No MeSH data available.


Related in: MedlinePlus

MALDI-TOFF spectra of the free peptide RGD (a), the free lipid (bupper panel) and the coupled RGD to mPEG in lipopeptide (blower panel).
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Fig1: MALDI-TOFF spectra of the free peptide RGD (a), the free lipid (bupper panel) and the coupled RGD to mPEG in lipopeptide (blower panel).

Mentions: RGD-TSL and TSL consisted of the phospholipids DPPC, DSPC, DSPE-PEG2000 in a molar ratio (70:25:5). In the RGD-TSL formulation, RGD peptide was coupled to mPEG2000 and its coupling efficiency was analyzed by MALDI (Fig. 1), after which it was mixed with all the lipids in a molar ratio (70:25:5). Both formulations were prepared by lipid film hydration and extrusion method (28). Dox was loaded into the liposomes after extrusion by (NH4)2SO4 loading method (29). Liposomes were characterized by measuring size, polydisperity index (pdI), encapsulation efficiency and stability. All liposomes were ~85 nm after extrusion and with a pdI < 0.1. After Dox loading, liposomes retained their small size and a low pdI. Encapsulation efficiency of both formulations was >95%. Liposomes were comparably stable after 1 h of incubation at 37°C in 99% FCS. TSL contained 92% ± 1.8 of the entrapped Dox after 1 h at 37°C whereas RGD-TSL contained 85% ± 2.3 (data not shown).Fig. 1


Enhanced Specificity and Drug Delivery in Tumors by cRGD - Anchoring Thermosensitive Liposomes.

Dicheva BM, Ten Hagen TL, Seynhaeve AL, Amin M, Eggermont AM, Koning GA - Pharm. Res. (2015)

MALDI-TOFF spectra of the free peptide RGD (a), the free lipid (bupper panel) and the coupled RGD to mPEG in lipopeptide (blower panel).
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4628091&req=5

Fig1: MALDI-TOFF spectra of the free peptide RGD (a), the free lipid (bupper panel) and the coupled RGD to mPEG in lipopeptide (blower panel).
Mentions: RGD-TSL and TSL consisted of the phospholipids DPPC, DSPC, DSPE-PEG2000 in a molar ratio (70:25:5). In the RGD-TSL formulation, RGD peptide was coupled to mPEG2000 and its coupling efficiency was analyzed by MALDI (Fig. 1), after which it was mixed with all the lipids in a molar ratio (70:25:5). Both formulations were prepared by lipid film hydration and extrusion method (28). Dox was loaded into the liposomes after extrusion by (NH4)2SO4 loading method (29). Liposomes were characterized by measuring size, polydisperity index (pdI), encapsulation efficiency and stability. All liposomes were ~85 nm after extrusion and with a pdI < 0.1. After Dox loading, liposomes retained their small size and a low pdI. Encapsulation efficiency of both formulations was >95%. Liposomes were comparably stable after 1 h of incubation at 37°C in 99% FCS. TSL contained 92% ± 1.8 of the entrapped Dox after 1 h at 37°C whereas RGD-TSL contained 85% ± 2.3 (data not shown).Fig. 1

Bottom Line: Cytotoxic effect of TSL and RGD-TSL was studied on B16Bl6 melanoma, B16F10 melanoma and HUVEC.High resolution intravital microscopy demonstrated specific accumulation of RGD-TSL to the tumor vasculature.Moreover, application of hyperthermia resulted in massive drug release from RGD-TSL.

View Article: PubMed Central - PubMed

Affiliation: Laboratory Experimental Surgical Oncology, Section Surgical Oncology Department of Surgery, Erasmus MC Cancer Center, Rotterdam, The Netherlands. b.dicheva@erasmusmc.nl.

ABSTRACT

Purpose: To develop RGD-targeted thermosensitive liposomes with increased tumor retention, improving drug release efficiency upon mild hyperthermia (HT) in both tumor and angiogenic endothelial cells.

Methods: Standard termosensitive liposomes (TSL) and TSL containing a cyclic Arg-Gly-Asp (cRGD) pentapeptide with the sequence Arg-Cys-D-Phe-Asp-Gly (RGDf[N-Met]C) were synthetized, loaded with Dox and characterized. Temperature- and time-dependent drug release profiles were assessed by fluorometry. Intracellular Dox delivery was studied by flow cytometry and confocal microscopy. Cytotoxic effect of TSL and RGD-TSL was studied on B16Bl6 melanoma, B16F10 melanoma and HUVEC. Intravital microscopy was performed on B16Bl6 tumors implanted in dorsal-skin fold window-bearing mice. Pharmacokinetic and biodistribution of Dox-TSL and Dox-RGD-TSL were followed in B16Bl6 tumor bearing mice upon normothermia or initial hyperthermia conditions.

Results: DLS and cryo-TEM revealed particle homogeneity and size of around 85 nm. Doxorubicin loading efficiency was >95%as assessed by spectrofluorometry. Flow cytometry and confocal microscopy showed a specific uptake of RGD-TSL by melanoma and endothelial cells when compared to TSL and an increased doxorubicin delivery. High resolution intravital microscopy demonstrated specific accumulation of RGD-TSL to the tumor vasculature. Moreover, application of hyperthermia resulted in massive drug release from RGD-TSL. Biodistribution studies showed that initial hyperthermia increases Dox uptake in tumors from TSL and RGD-TSL.

Conclusion: RGD-TSL have potency to increase drug efficacy due to higher uptake by tumor and angiogenic endothelial cells in combination with heat-triggered drug release.

No MeSH data available.


Related in: MedlinePlus