Hybrid Albumin-Decorated Lipid-Nanocarrier-Mediated Delivery of Polyphenol-Rich Sambucus nigra L. in a Potential Multiple Antitumoural Therapy

Abstract

The current research attempted to address the suitability of bioactive Sambucus nigra extract entrapped in albumin-decorated nanostructured lipid carriers (NLCs) as a promising "adjuvant" in improving tumour penetration for multiple antitumour therapy. The new hybrid albumin-decorated NLCs were characterised based on, e.g., the particle size, zeta electrokinetic potential, SambucusN entrapment efficiency, and fluorescence spectroscopy and tested for different formulation parameters. The antioxidant activity of NLC-SambucusN was significantly enhanced by a bovine serum albumin (BSA) polymer coating. According to the real-time cell analysis (RTCA) results, NLC-I-SambucusN-BSA behaved similarly to the chemotherapeutic drug, cisplatin, with cell viability for LoVo tumour cells of 21.81 ± 1.18%. The new albumin-NLC-SambucusN arrested cancer cells in G1 and G2 cycles and intensified the apoptosis process in both early and late phases. An advanced induction, over 50% apoptosis in LoVo colon cells, was registered for 50 μg/mL of NLC-II-SambucusN-BSA, a fourfold increase compared to that of untreated cells. RTCA and flow cytometry results showed that concentrations of the hybrid NLC-SambucusN up to 50 μg/mL do not affect the proliferation of normal HUVEC cells. This approach provides insightful information regarding the involvement of phytochemicals in future therapeutic strategies. Albumin-decorated NLCs can be considered a noteworthy strategy to be connected to antitumour therapeutic protocols.

Keywords: Sambucus -nigra; antioxidant effect; apoptosis process; cell cycle arrest; phytochemical entrapment; protein–lipid nanocarriers.

Figure 5

Effect of the conventional NLC-I and -II and hybrid BSA-coated NLC-I and -II, loaded with SambucusN extract, on the cell viability of the following: (A) normal HUVEC cells; (B) LoVo colon cancer cells; (C) MCF-7 breast cancer cells; and (D) SKOV-3 ovary cancer cells after 24 h and 48 h of incubation with developed NLC and chemotherapy drugs, cisplatin and doxorubicin. All experiments were performed in triplicate. * p < 0.05; ** p < 0.005; *** p < 0.0005. Data are expressed as mean ± SD, n = 3 NLCI/II vs. other groups.

Figure 6

Real-time cell analysis (RTCA) analysis recorded on normal HUVEC cells (A), normal LoVo colon cancer cells (B), normal MCF-7 breast cancer cells (C), and normal SKOV-3 ovarian cancer cells (D) for conventional NLC-I and -II and hybrid BSA-coated NLC-I and -II, with and without SambucusN extract.

Figure 6

Real-time cell analysis (RTCA) analysis recorded on normal HUVEC cells (A), normal LoVo colon cancer cells (B), normal MCF-7 breast cancer cells (C), and normal SKOV-3 ovarian cancer cells (D) for conventional NLC-I and -II and hybrid BSA-coated NLC-I and -II, with and without SambucusN extract.

Figure 6

Real-time cell analysis (RTCA) analysis recorded on normal HUVEC cells (A), normal LoVo colon cancer cells (B), normal MCF-7 breast cancer cells (C), and normal SKOV-3 ovarian cancer cells (D) for conventional NLC-I and -II and hybrid BSA-coated NLC-I and -II, with and without SambucusN extract.

Figure 6

Real-time cell analysis (RTCA) analysis recorded on normal HUVEC cells (A), normal LoVo colon cancer cells (B), normal MCF-7 breast cancer cells (C), and normal SKOV-3 ovarian cancer cells (D) for conventional NLC-I and -II and hybrid BSA-coated NLC-I and -II, with and without SambucusN extract.

Figure 7

Apoptotic process induced by various categories of NLC on different cancer cells: (A) normal HUVEC cells; (B) LoVo colon tumour cells; (C) MCF-7 breast tumour cells; (D) SKOV-3 ovary tumour cells. All experiments were performed in triplicate. * p < 0.05. Data are expressed as mean ± SD, n = 3 NLCI/II vs. other groups.

Figure 7

Apoptotic process induced by various categories of NLC on different cancer cells: (A) normal HUVEC cells; (B) LoVo colon tumour cells; (C) MCF-7 breast tumour cells; (D) SKOV-3 ovary tumour cells. All experiments were performed in triplicate. * p < 0.05. Data are expressed as mean ± SD, n = 3 NLCI/II vs. other groups.

Figure 8

The influence of the conventional NLC and hybrid BSA-coated NLC on cell cycle in tumour cells: (A) normal HUVEC cells; (B) LoVo colon tumour cells; (C) MCF-7 breast tumour cells; (D) SKOV-3 ovary tumour cells.

Figure 8

The influence of the conventional NLC and hybrid BSA-coated NLC on cell cycle in tumour cells: (A) normal HUVEC cells; (B) LoVo colon tumour cells; (C) MCF-7 breast tumour cells; (D) SKOV-3 ovary tumour cells.

Figure 1

Different quality attributes of lipid and hybrid BSA−lipid nanocarriers hosting SambucusN extract: main diameter size (A), electrokinetic potential (B), entrapment efficiency (C), and FT-ICR MS/MS of SambucusN (D). All experiments were performed in triplicate. * p < 0.05; ** p < 0.005; *** p < 0.0005. Data are expressed as mean ± SD, n = 3 NLCI/II vs. other groups.

Figure 1

Different quality attributes of lipid and hybrid BSA−lipid nanocarriers hosting SambucusN extract: main diameter size (A), electrokinetic potential (B), entrapment efficiency (C), and FT-ICR MS/MS of SambucusN (D). All experiments were performed in triplicate. * p < 0.05; ** p < 0.005; *** p < 0.0005. Data are expressed as mean ± SD, n = 3 NLCI/II vs. other groups.

Figure 2

Comparative ATR-FTIR spectra of coated and uncoated NLC-I−SambucusN (A) and NLC-II−SambucusN (B). Details on ATR−FTIR spectra of NLC-I (C) and II−SambucusN (D) coated and uncoated with protein.

Figure 3

Comparative fluorescence spectra of the conventional NLC and hybrid BSA-decorated NLC, loaded or free of SambucusN extract.

Figure 4

Antioxidant activity of NLC formulations: scavenging activity of ABTS radical cations by NLC-I and -II SambucusN with and without BSA polymer (A); dose−response curves and IC50 values of BSA-coated NLC (B). All experiments were performed in triplicate. * p < 0.05; ** p < 0.005.