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Virosomes of hepatitis B virus envelope L proteins containing doxorubicin: synergistic enhancement of human liver-specific antitumor growth activity by radiotherapy.

Liu Q, Jung J, Somiya M, Iijima M, Yoshimoto N, Niimi T, Maturana AD, Shin SH, Jeong SY, Choi EK, Kuroda S - Int J Nanomedicine (2015)

Bottom Line: The growth of target cell-derived tumors was retarded effectively and specifically.Next, the combination of high dose (10.0 mg/kg as DOX, once) with tumor-specific radiotherapy (3 Gy, once after 2 hours) exhibited the most effective antitumor growth activity in mice harboring target cell-derived tumors.These results demonstrated that the HBV-based virosomes containing DOX could be an effective antitumor nanomedicine specific to human hepatic tissues, especially in combination with radiotherapy.

View Article: PubMed Central - PubMed

Affiliation: Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, Japan ; The Institute of Scientific and Industrial Research, Osaka University, Ibaraki, Japan.

ABSTRACT
Bionanocapsules (BNCs) are hollow nanoparticles consisting of hepatitis B virus (HBV) envelope L proteins and have been shown to deliver drugs and genes specifically to human hepatic tissues by utilizing HBV-derived infection machinery. The complex of BNCs with liposomes (LPs), the BNC-LP complexes (a LP surrounded by BNCs in a rugged spherical form), could also become active targeting nanocarriers by the BNC function. In this study, under acidic conditions and high temperature, BNCs were found to fully fuse with LPs (smooth-surfaced spherical form), deploying L proteins with a membrane topology similar to that of BNCs (ie, virosomes displaying L proteins). Doxorubicin (DOX) was efficiently encapsulated via the remote loading method at 14.2%±1.0% of total lipid weight (mean ± SD, n=3), with a capsule size of 118.2±4.7 nm and a ζ-potential of -51.1±1.0 mV (mean ± SD, n=5). When mammalian cells were exposed to the virosomes, the virosomes showed strong cytotoxicity in human hepatic cells (target cells of BNCs), but not in human colon cancer cells (nontarget cells of BNCs), whereas LPs containing DOX and DOXOVES (structurally stabilized PEGylated LPs containing DOX) did not show strong cytotoxicity in either cell type. Furthermore, the virosomes preferentially delivered DOX to the nuclei of human hepatic cells. Xenograft mice harboring either target or nontarget cell-derived tumors were injected twice intravenously with the virosomes containing DOX at a low dose (2.3 mg/kg as DOX, 5 days interval). The growth of target cell-derived tumors was retarded effectively and specifically. Next, the combination of high dose (10.0 mg/kg as DOX, once) with tumor-specific radiotherapy (3 Gy, once after 2 hours) exhibited the most effective antitumor growth activity in mice harboring target cell-derived tumors. These results demonstrated that the HBV-based virosomes containing DOX could be an effective antitumor nanomedicine specific to human hepatic tissues, especially in combination with radiotherapy.

No MeSH data available.


Related in: MedlinePlus

In vitro cytotoxicity assay.Notes: Huh7 (A) or WiDr (B) cells were treated with DOX (blue diamonds), DOXOVES (purple triangles), LP–DOX complexes (green circles), and virosomes containing DOX (red squares) at 37°C for 6 hours. Cell viability was measured using a commercial WST-8 assay reagent. Error bars represent standard deviation. t-test, **P<0.01, *P<0.05.Abbreviations: DOX, doxorubicin; LP, liposome; DOXOVES, structurally stabilized PEGylated LPs containing DOX; WST-8, 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt.
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f5-ijn-10-4159: In vitro cytotoxicity assay.Notes: Huh7 (A) or WiDr (B) cells were treated with DOX (blue diamonds), DOXOVES (purple triangles), LP–DOX complexes (green circles), and virosomes containing DOX (red squares) at 37°C for 6 hours. Cell viability was measured using a commercial WST-8 assay reagent. Error bars represent standard deviation. t-test, **P<0.01, *P<0.05.Abbreviations: DOX, doxorubicin; LP, liposome; DOXOVES, structurally stabilized PEGylated LPs containing DOX; WST-8, 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt.

Mentions: Huh7 cells (target cells) and WiDr cells (nontarget cells) were incubated with DOX, LP–DOX complexes, DOXOVES (commercial DOX-containing PEGylated LPs), and virosomes containing DOX at DOX concentrations of 0.195–50 μg/mL for 6 hours, and then subjected to WST-8 assay. In Huh7 cells, the virosomes containing DOX showed higher cytotoxicity than the LP–DOX complexes and DOXOVES (Figure 5A). The IC50 value (50% inhibitory concentration for cell growth) of each drug was ~4.0 μg/mL (DOX), ~12.5 μg/mL (virosomes containing DOX), ~25.0 μg/mL (LP-DOX), and ~46.0 μg/mL (DOXOVES). In WiDr cells (Figure 5B), while DOX showed strong cytotoxicity (~2.5 μg/mL), the other drugs showed low cytotoxicity (>100 μg/mL). These results demonstrated that the full fusion with BNCs conferred LP–DOX complexes with the targeting ability that is specific to human hepatic cells.


Virosomes of hepatitis B virus envelope L proteins containing doxorubicin: synergistic enhancement of human liver-specific antitumor growth activity by radiotherapy.

Liu Q, Jung J, Somiya M, Iijima M, Yoshimoto N, Niimi T, Maturana AD, Shin SH, Jeong SY, Choi EK, Kuroda S - Int J Nanomedicine (2015)

In vitro cytotoxicity assay.Notes: Huh7 (A) or WiDr (B) cells were treated with DOX (blue diamonds), DOXOVES (purple triangles), LP–DOX complexes (green circles), and virosomes containing DOX (red squares) at 37°C for 6 hours. Cell viability was measured using a commercial WST-8 assay reagent. Error bars represent standard deviation. t-test, **P<0.01, *P<0.05.Abbreviations: DOX, doxorubicin; LP, liposome; DOXOVES, structurally stabilized PEGylated LPs containing DOX; WST-8, 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4487236&req=5

f5-ijn-10-4159: In vitro cytotoxicity assay.Notes: Huh7 (A) or WiDr (B) cells were treated with DOX (blue diamonds), DOXOVES (purple triangles), LP–DOX complexes (green circles), and virosomes containing DOX (red squares) at 37°C for 6 hours. Cell viability was measured using a commercial WST-8 assay reagent. Error bars represent standard deviation. t-test, **P<0.01, *P<0.05.Abbreviations: DOX, doxorubicin; LP, liposome; DOXOVES, structurally stabilized PEGylated LPs containing DOX; WST-8, 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt.
Mentions: Huh7 cells (target cells) and WiDr cells (nontarget cells) were incubated with DOX, LP–DOX complexes, DOXOVES (commercial DOX-containing PEGylated LPs), and virosomes containing DOX at DOX concentrations of 0.195–50 μg/mL for 6 hours, and then subjected to WST-8 assay. In Huh7 cells, the virosomes containing DOX showed higher cytotoxicity than the LP–DOX complexes and DOXOVES (Figure 5A). The IC50 value (50% inhibitory concentration for cell growth) of each drug was ~4.0 μg/mL (DOX), ~12.5 μg/mL (virosomes containing DOX), ~25.0 μg/mL (LP-DOX), and ~46.0 μg/mL (DOXOVES). In WiDr cells (Figure 5B), while DOX showed strong cytotoxicity (~2.5 μg/mL), the other drugs showed low cytotoxicity (>100 μg/mL). These results demonstrated that the full fusion with BNCs conferred LP–DOX complexes with the targeting ability that is specific to human hepatic cells.

Bottom Line: The growth of target cell-derived tumors was retarded effectively and specifically.Next, the combination of high dose (10.0 mg/kg as DOX, once) with tumor-specific radiotherapy (3 Gy, once after 2 hours) exhibited the most effective antitumor growth activity in mice harboring target cell-derived tumors.These results demonstrated that the HBV-based virosomes containing DOX could be an effective antitumor nanomedicine specific to human hepatic tissues, especially in combination with radiotherapy.

View Article: PubMed Central - PubMed

Affiliation: Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, Japan ; The Institute of Scientific and Industrial Research, Osaka University, Ibaraki, Japan.

ABSTRACT
Bionanocapsules (BNCs) are hollow nanoparticles consisting of hepatitis B virus (HBV) envelope L proteins and have been shown to deliver drugs and genes specifically to human hepatic tissues by utilizing HBV-derived infection machinery. The complex of BNCs with liposomes (LPs), the BNC-LP complexes (a LP surrounded by BNCs in a rugged spherical form), could also become active targeting nanocarriers by the BNC function. In this study, under acidic conditions and high temperature, BNCs were found to fully fuse with LPs (smooth-surfaced spherical form), deploying L proteins with a membrane topology similar to that of BNCs (ie, virosomes displaying L proteins). Doxorubicin (DOX) was efficiently encapsulated via the remote loading method at 14.2%±1.0% of total lipid weight (mean ± SD, n=3), with a capsule size of 118.2±4.7 nm and a ζ-potential of -51.1±1.0 mV (mean ± SD, n=5). When mammalian cells were exposed to the virosomes, the virosomes showed strong cytotoxicity in human hepatic cells (target cells of BNCs), but not in human colon cancer cells (nontarget cells of BNCs), whereas LPs containing DOX and DOXOVES (structurally stabilized PEGylated LPs containing DOX) did not show strong cytotoxicity in either cell type. Furthermore, the virosomes preferentially delivered DOX to the nuclei of human hepatic cells. Xenograft mice harboring either target or nontarget cell-derived tumors were injected twice intravenously with the virosomes containing DOX at a low dose (2.3 mg/kg as DOX, 5 days interval). The growth of target cell-derived tumors was retarded effectively and specifically. Next, the combination of high dose (10.0 mg/kg as DOX, once) with tumor-specific radiotherapy (3 Gy, once after 2 hours) exhibited the most effective antitumor growth activity in mice harboring target cell-derived tumors. These results demonstrated that the HBV-based virosomes containing DOX could be an effective antitumor nanomedicine specific to human hepatic tissues, especially in combination with radiotherapy.

No MeSH data available.


Related in: MedlinePlus