Biomimetic cancer cell membrane engineered lipid nanoparticles for enhanced chemotherapy of homologous malignant tumor

Abstract

Background: The advancement of biomimetic drug delivery systems designed for biomedical applications has attracted considerable attention from researchers in recent years. A particularly noteworthy approach involves the use of various cell membranes, which can impart distinctive functionalities to the nanoparticles, including specific recognition of target cells, prolonged circulation within the bloodstream, and enhanced ability to evade the immune system, as surface coatings on nanoparticles. This innovative strategy has positioned cell membrane-coated nanoparticles (CMCNPs) as a promising framework for addressing a wide range of diseases more effectively.

Methods: In the current investigation, lipid nanoparticles were specifically engineered using glioblastoma cell membrane (GBMM) coatings, termed as LNPs/D@GBMM, to serve as targeted nanotheranostics against homologous malignant glioblastoma (GBM). The physicochemical properties of LNPs/D@GBMM were investigated in terms of particle size, morphology, drug loading (DL), drug release behavior and so on. Homologous cellular uptake was evaluated by confocal laser scanning microscopy (CLSM). Cell cytotoxicity was evaluated by MTT assay. Moreover, the bio-distribution of CMCNPs in vivo was investigated via the near-infrared (NIR) fluorescence imaging technique, and the anti-tumor effect in vivo was evaluated in xenografted nude mice.

Results: Compared to non-targeted lipid nanoparticles, LNPs/D@GBMM exhibited superior cytotoxic effects against homologous tumor cells. In addition, fluorescence imaging of targeted tumor cells treated with LNPs/D@GBMM indicated a marked increase in cell internalization, and improved fluorescence distribution in vivo. LNPs/D@GBMM finally produced an excellent tumor suppression effect on homologous tumors.

Conclusion: The robust platform established by CMCNPs leveraging the inherent characteristics of homologous tumor cell membranes, is expected to facilitate systemic delivery of therapeutic agents specifically aimed at treating tumors, thus advancing the efficacy of cancer therapy in clinical settings.

Keywords: Biomimetic drug delivery systems; Cell membrane; Glioblastoma; Homologous tumor; Lipid nanoparticles.

Fig. 1

Characterization of membrane-coated nanoparticles. a SDS-PAGE analysis. b Western blotting analysis of U87 MG cells, GBMM, LNPs/D@GBMM nanoparticles. c TEM images of LNPs, LNPs/D and LNPs/D@GBMM nanoparticles (Scale bar = 100 nm)

Fig. 2

DOX release behaviors of free DOX·HCl, LNPs/D and LNPs/D@GBMM formulations under (a) pH 7.4 and (b) pH 6.8 PBS solutions within 48 h (n = 3)

Fig. 3

Cellular uptake. Intracellular uptake of free DOX·HCl, LNPs/D, LNPs/D@GBMM in (a) U87 MG cells and (b) MG 63 cells for 4 h, 8 h and 24 h determined by CLSM (scale bar = 30 μm)

Fig. 4

Cytotoxicity. Cell survival rates of free DOX·HCl, LNPs/D, LNPs/D@GBMM against (a) U87 MG cells and (b) MG 63 cells, respectively (n = 6)

Fig. 5

Cell migration suppression. a The wound width was observed and measured by a microscope 1 h after wounding, and images of wound healing assays of U87 MG cells treated with free DOX·HCl, LNPs/D and LNPs/D@GBMM nanoparticles (containing 1.5 µg/mL of DOX) for 24 h. b Semi-quantitative results of wound healing percentages (n = 3)

Fig. 6

Bio-distribution in vivo. a In vivo fluorescence imaging of free DOX·HCl, LNPs/D, LNPs/D@GBMM in subcutaneous GBM models 12 h and 24 h after i.v. administration. b Ex vivo imaging of free DOX·HCl, LNPs/D, LNPs/D@GBMM nanoparticles in tumor cells and vital organs after i.v. injection for 24 h. c Fluorescence distribution of free DOX·HCl, LNPs/D, LNPs/D@GBMM in tumor tissues. d The fluorescence semi-quantitative analysis was evaluated by Image J (n = 3)

Fig. 7

Anti-tumor efficacy of LNPs/D@GBMM nanoparticles in vivo. a Tumor growth curves of saline group, free DOX·HCl group, LNPs/D group and LNPs/D@GBMM group. b Images of excised tumor tissues from all experimental groups, c average tumor weight for each experimental group at the end of treatment (n = 5). d Body weight curves of all experimental groups measured every two days after the first injection (n = 5). e H&E staining of tumor sections and immunohistochemical analysis of tumor sections stained with Bax, Bcl-2, CD31 and Ki67 (brown). Magnification: 200× (Scale bar = 100 μm)

Fig. 8

Safety evaluation in vivo. Studies of the long-term toxicity of different formational drugs in the heart, liver, spleen, lung and kidney using H&E staining. Magnification: 200× (Scale bar = 100 μm)