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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

Stability of virosomes containing DOX.Notes: The virosomes containing DOX were kept in outer aqueous phase buffer at 4°C (red) and 37°C (blue) for 8 weeks. The relative amount of DOX (A) and particle size (B) were measured at 0, 1, 2, 3, 4, and 8 weeks. The virosomes containing DOX were kept in outer aqueous phase buffer at 4°C and 37°C for 8 weeks. The relative amount of DOX (C) and particle size (D) were measured at 0, 1, 2, 4, 5, 6, and 8 days. The virosomes containing DOX were kept in 50% FBS at 37°C for 8 days. Error bars represent standard deviation (n=3).Abbreviations: DOX, doxorubicin; FBS, fetal bovine serum.
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f4-ijn-10-4159: Stability of virosomes containing DOX.Notes: The virosomes containing DOX were kept in outer aqueous phase buffer at 4°C (red) and 37°C (blue) for 8 weeks. The relative amount of DOX (A) and particle size (B) were measured at 0, 1, 2, 3, 4, and 8 weeks. The virosomes containing DOX were kept in outer aqueous phase buffer at 4°C and 37°C for 8 weeks. The relative amount of DOX (C) and particle size (D) were measured at 0, 1, 2, 4, 5, 6, and 8 days. The virosomes containing DOX were kept in 50% FBS at 37°C for 8 days. Error bars represent standard deviation (n=3).Abbreviations: DOX, doxorubicin; FBS, fetal bovine serum.

Mentions: The stability of virosomes containing DOX was evaluated in 10 mM HEPES buffer (pH 7.4) containing 100 mM NaCl and 3.4% sucrose (outer aqueous phase buffer) at 4°C or 37°C. Based on the relative amount of DOX remaining, the virosomes were found to retain DOX long-term (8 weeks) at 4°C rather than at 37°C (Figure 4A), while the diameter of the virosomes increased slightly (Figure 4B). Thus, the structure of virosomes containing DOX was well maintained in the outer aqueous phase buffer at 4°C for at least 8 weeks. Furthermore, the virosomes containing DOX were stored in 50% FBS (containing active complements) at 37°C for 8 days. More than 70% of the incorporated DOX was retained in the virosomes during the first 2 days, whereas about 80% of the incorporated DOX was gradually released from the virosomes during another 4 days (Figure 4C). The diameter of the virosomes containing DOX was significantly increased within 6 days (Figure 4D). When using the previous version of BNC–LP–DOX complexes, the conjugation of BNCs could enhance the retention time of DOX in the bloodstream more efficiently.18 Thus, the virosomes containing DOX would harbor comparable level of stability for use in vivo.


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)

Stability of virosomes containing DOX.Notes: The virosomes containing DOX were kept in outer aqueous phase buffer at 4°C (red) and 37°C (blue) for 8 weeks. The relative amount of DOX (A) and particle size (B) were measured at 0, 1, 2, 3, 4, and 8 weeks. The virosomes containing DOX were kept in outer aqueous phase buffer at 4°C and 37°C for 8 weeks. The relative amount of DOX (C) and particle size (D) were measured at 0, 1, 2, 4, 5, 6, and 8 days. The virosomes containing DOX were kept in 50% FBS at 37°C for 8 days. Error bars represent standard deviation (n=3).Abbreviations: DOX, doxorubicin; FBS, fetal bovine serum.
© Copyright Policy
Related In: Results  -  Collection

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

f4-ijn-10-4159: Stability of virosomes containing DOX.Notes: The virosomes containing DOX were kept in outer aqueous phase buffer at 4°C (red) and 37°C (blue) for 8 weeks. The relative amount of DOX (A) and particle size (B) were measured at 0, 1, 2, 3, 4, and 8 weeks. The virosomes containing DOX were kept in outer aqueous phase buffer at 4°C and 37°C for 8 weeks. The relative amount of DOX (C) and particle size (D) were measured at 0, 1, 2, 4, 5, 6, and 8 days. The virosomes containing DOX were kept in 50% FBS at 37°C for 8 days. Error bars represent standard deviation (n=3).Abbreviations: DOX, doxorubicin; FBS, fetal bovine serum.
Mentions: The stability of virosomes containing DOX was evaluated in 10 mM HEPES buffer (pH 7.4) containing 100 mM NaCl and 3.4% sucrose (outer aqueous phase buffer) at 4°C or 37°C. Based on the relative amount of DOX remaining, the virosomes were found to retain DOX long-term (8 weeks) at 4°C rather than at 37°C (Figure 4A), while the diameter of the virosomes increased slightly (Figure 4B). Thus, the structure of virosomes containing DOX was well maintained in the outer aqueous phase buffer at 4°C for at least 8 weeks. Furthermore, the virosomes containing DOX were stored in 50% FBS (containing active complements) at 37°C for 8 days. More than 70% of the incorporated DOX was retained in the virosomes during the first 2 days, whereas about 80% of the incorporated DOX was gradually released from the virosomes during another 4 days (Figure 4C). The diameter of the virosomes containing DOX was significantly increased within 6 days (Figure 4D). When using the previous version of BNC–LP–DOX complexes, the conjugation of BNCs could enhance the retention time of DOX in the bloodstream more efficiently.18 Thus, the virosomes containing DOX would harbor comparable level of stability for use in vivo.

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