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Doxorubicin-incorporated polymeric micelles composed of dextran-b-poly(DL-lactide-co-glycolide) copolymer.

Jeong YI, Kim do H, Chung CW, Yoo JJ, Choi KH, Kim CH, Ha SH, Kang DH - Int J Nanomedicine (2011)

Bottom Line: To investigate the in vitro anticancer effects of the polymeric micelles, doxorubicin-resistant HuCC-T1 cells were treated with a very high concentration of doxorubicin.Furthermore, in flow cytometric analysis, fluorescence intensity of polymeric micelles was almost twice as high than with free doxorubicin.DexbLG polymeric micelles incorporating doxorubicin are promising vehicles for antitumor drug targeting.

View Article: PubMed Central - PubMed

Affiliation: National Research and Development Center for Hepatobiliary Cancer, Pusan National University Yangsan Hospital, Yangsan, Republic of Korea.

ABSTRACT

Background: Polymeric micelles using amphiphilic macromolecules are promising vehicles for antitumor targeting. In this study, we prepared anticancer agent-incorporated polymeric micelles using novel block copolymer.

Methods: We synthesized a block copolymer composed of dextran and poly (DL-lactide-co-glycolide) (DexbLG) for antitumor drug delivery. Doxorubicin was selected as the anticancer drug, and was incorporated into polymeric micelles by dialysis. Polymeric micelles were observed by transmission electron microscopy to be spherical and smaller than 100 nm, with a narrow size distribution. The particle size of doxorubicin-incorporated polymeric micelles increased with increasing drug content. Higher initial drug feeding also increased the drug content.

Results: During the drug-release study, an initial burst release of doxorubicin was observed for 10 hours, and doxorubicin was continuously released over 4 days. To investigate the in vitro anticancer effects of the polymeric micelles, doxorubicin-resistant HuCC-T1 cells were treated with a very high concentration of doxorubicin. In an antiproliferation study, the polymeric micelles showed higher cytotoxicity to doxorubicin-resistant HuCC-T1 cells than free doxorubicin, indicating that the polymeric micelles were effectively engulfed by tumor cells, while free doxorubicin hardly penetrated the tumor cell membrane. On confocal laser scanning microscopy, free doxorubicin expressed very weak fluorescence intensity, while the polymeric micelles expressed strong red fluorescence. Furthermore, in flow cytometric analysis, fluorescence intensity of polymeric micelles was almost twice as high than with free doxorubicin.

Conclusion: DexbLG polymeric micelles incorporating doxorubicin are promising vehicles for antitumor drug targeting.

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Flow cytometric analysis of doxorubicin-sensitive and doxorubicin-resistant HuCC-T1 cells. HuCCT1 cells were exposed to doxorubicin or polymeric micelles (equivalent concentration of doxorubicin 1 μg/mL) for 1 hour. 1 × 106 cells were used for FACScan analysis. Doxorubicin-sensitive cells: (A) control, (B) doxorubicin, (C) polymeric micelles, (D) relative fluorescence intensity. Doxorubicin-resistant cells: (E) control; (F) doxorubicin; (G) polymeric micelles; (H) relative fluorescence intensity. The values were averaged over four separate experiments.Notes: *P < 0.005; **P < 0.0001; ***P < 0.0001; ****P < 0.0001.
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f10-ijn-6-1415: Flow cytometric analysis of doxorubicin-sensitive and doxorubicin-resistant HuCC-T1 cells. HuCCT1 cells were exposed to doxorubicin or polymeric micelles (equivalent concentration of doxorubicin 1 μg/mL) for 1 hour. 1 × 106 cells were used for FACScan analysis. Doxorubicin-sensitive cells: (A) control, (B) doxorubicin, (C) polymeric micelles, (D) relative fluorescence intensity. Doxorubicin-resistant cells: (E) control; (F) doxorubicin; (G) polymeric micelles; (H) relative fluorescence intensity. The values were averaged over four separate experiments.Notes: *P < 0.005; **P < 0.0001; ***P < 0.0001; ****P < 0.0001.

Mentions: To investigate the antitumor activity of free doxorubicin and DexbLG polymeric micelles incorporating doxorubicin, polymeric micelles incorporating doxorubicin were tested with doxorubicin-sensitive HuCC-T1 cells and doxorubicin-resistant HuCC-T1 cells. For this test, doxorubicin-resistant HuCC-T1 cells were exposed to doxorubicin 0.0001 μg/mL to 0.1 μg/mL for several months. As shown in Figure 7A, free doxorubicin and polymeric micelles incorporating doxorubicin showed dose-dependent growth inhibition of doxorubicin-sensitive HuCC-T1 cells. Cell viability on exposure to free doxorubicin and to polymeric micelles incorporating doxorubicin was not significantly different. However, polymeric micelles incorporating doxorubicin showed higher antitumor cytotoxicity against doxorubicin-resistant HuCC-T1 cells, as shown in Figure 7B. As shown in Figure 7C, the empty polymeric micelles did not significantly inhibit tumor cell growth. The IC50 of polymeric micelles against doxorubicin-resistant HuCC-T1 cells was significantly lower than that for free doxorubicin, as shown in Figure 8B, but for doxorubicin-sensitive cells was not significantly different. To verify these results, HuCC-T1 cells were observed using confocal laser scanning microscopy because doxorubicin has strong red fluorescence, ie, tumor cells will reveal red fluorescence when doxorubicin enters the cell. Figure 9 supports the growth inhibition data. When doxorubicin-resistant cells were treated with free doxorubicin, HuCC-T1 cells revealed very weak red fluorescence intensity, while the polymeric micelles showed strong red fluorescence intensity. Figure 10 also supported these results, ie, polymeric micelle treatment showed higher fluorescence intensity than free doxorubicin treatment. These results showed that DexbLG polymeric micelles incorporating doxorubicin are superior candidates for anticancer drug targeting.


Doxorubicin-incorporated polymeric micelles composed of dextran-b-poly(DL-lactide-co-glycolide) copolymer.

Jeong YI, Kim do H, Chung CW, Yoo JJ, Choi KH, Kim CH, Ha SH, Kang DH - Int J Nanomedicine (2011)

Flow cytometric analysis of doxorubicin-sensitive and doxorubicin-resistant HuCC-T1 cells. HuCCT1 cells were exposed to doxorubicin or polymeric micelles (equivalent concentration of doxorubicin 1 μg/mL) for 1 hour. 1 × 106 cells were used for FACScan analysis. Doxorubicin-sensitive cells: (A) control, (B) doxorubicin, (C) polymeric micelles, (D) relative fluorescence intensity. Doxorubicin-resistant cells: (E) control; (F) doxorubicin; (G) polymeric micelles; (H) relative fluorescence intensity. The values were averaged over four separate experiments.Notes: *P < 0.005; **P < 0.0001; ***P < 0.0001; ****P < 0.0001.
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Related In: Results  -  Collection

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f10-ijn-6-1415: Flow cytometric analysis of doxorubicin-sensitive and doxorubicin-resistant HuCC-T1 cells. HuCCT1 cells were exposed to doxorubicin or polymeric micelles (equivalent concentration of doxorubicin 1 μg/mL) for 1 hour. 1 × 106 cells were used for FACScan analysis. Doxorubicin-sensitive cells: (A) control, (B) doxorubicin, (C) polymeric micelles, (D) relative fluorescence intensity. Doxorubicin-resistant cells: (E) control; (F) doxorubicin; (G) polymeric micelles; (H) relative fluorescence intensity. The values were averaged over four separate experiments.Notes: *P < 0.005; **P < 0.0001; ***P < 0.0001; ****P < 0.0001.
Mentions: To investigate the antitumor activity of free doxorubicin and DexbLG polymeric micelles incorporating doxorubicin, polymeric micelles incorporating doxorubicin were tested with doxorubicin-sensitive HuCC-T1 cells and doxorubicin-resistant HuCC-T1 cells. For this test, doxorubicin-resistant HuCC-T1 cells were exposed to doxorubicin 0.0001 μg/mL to 0.1 μg/mL for several months. As shown in Figure 7A, free doxorubicin and polymeric micelles incorporating doxorubicin showed dose-dependent growth inhibition of doxorubicin-sensitive HuCC-T1 cells. Cell viability on exposure to free doxorubicin and to polymeric micelles incorporating doxorubicin was not significantly different. However, polymeric micelles incorporating doxorubicin showed higher antitumor cytotoxicity against doxorubicin-resistant HuCC-T1 cells, as shown in Figure 7B. As shown in Figure 7C, the empty polymeric micelles did not significantly inhibit tumor cell growth. The IC50 of polymeric micelles against doxorubicin-resistant HuCC-T1 cells was significantly lower than that for free doxorubicin, as shown in Figure 8B, but for doxorubicin-sensitive cells was not significantly different. To verify these results, HuCC-T1 cells were observed using confocal laser scanning microscopy because doxorubicin has strong red fluorescence, ie, tumor cells will reveal red fluorescence when doxorubicin enters the cell. Figure 9 supports the growth inhibition data. When doxorubicin-resistant cells were treated with free doxorubicin, HuCC-T1 cells revealed very weak red fluorescence intensity, while the polymeric micelles showed strong red fluorescence intensity. Figure 10 also supported these results, ie, polymeric micelle treatment showed higher fluorescence intensity than free doxorubicin treatment. These results showed that DexbLG polymeric micelles incorporating doxorubicin are superior candidates for anticancer drug targeting.

Bottom Line: To investigate the in vitro anticancer effects of the polymeric micelles, doxorubicin-resistant HuCC-T1 cells were treated with a very high concentration of doxorubicin.Furthermore, in flow cytometric analysis, fluorescence intensity of polymeric micelles was almost twice as high than with free doxorubicin.DexbLG polymeric micelles incorporating doxorubicin are promising vehicles for antitumor drug targeting.

View Article: PubMed Central - PubMed

Affiliation: National Research and Development Center for Hepatobiliary Cancer, Pusan National University Yangsan Hospital, Yangsan, Republic of Korea.

ABSTRACT

Background: Polymeric micelles using amphiphilic macromolecules are promising vehicles for antitumor targeting. In this study, we prepared anticancer agent-incorporated polymeric micelles using novel block copolymer.

Methods: We synthesized a block copolymer composed of dextran and poly (DL-lactide-co-glycolide) (DexbLG) for antitumor drug delivery. Doxorubicin was selected as the anticancer drug, and was incorporated into polymeric micelles by dialysis. Polymeric micelles were observed by transmission electron microscopy to be spherical and smaller than 100 nm, with a narrow size distribution. The particle size of doxorubicin-incorporated polymeric micelles increased with increasing drug content. Higher initial drug feeding also increased the drug content.

Results: During the drug-release study, an initial burst release of doxorubicin was observed for 10 hours, and doxorubicin was continuously released over 4 days. To investigate the in vitro anticancer effects of the polymeric micelles, doxorubicin-resistant HuCC-T1 cells were treated with a very high concentration of doxorubicin. In an antiproliferation study, the polymeric micelles showed higher cytotoxicity to doxorubicin-resistant HuCC-T1 cells than free doxorubicin, indicating that the polymeric micelles were effectively engulfed by tumor cells, while free doxorubicin hardly penetrated the tumor cell membrane. On confocal laser scanning microscopy, free doxorubicin expressed very weak fluorescence intensity, while the polymeric micelles expressed strong red fluorescence. Furthermore, in flow cytometric analysis, fluorescence intensity of polymeric micelles was almost twice as high than with free doxorubicin.

Conclusion: DexbLG polymeric micelles incorporating doxorubicin are promising vehicles for antitumor drug targeting.

Show MeSH
Related in: MedlinePlus