Hyaluronic acid-based nano drug delivery systems for breast cancer treatment: Recent advances

Abstract

Breast cancer (BC) is the most common malignancy among females worldwide, and high resistance to drugs and metastasis rates are the leading causes of death in BC patients. Releasing anti-cancer drugs precisely to the tumor site can improve the efficacy and reduce the side effects on the body. Natural polymers are attracting extensive interest as drug carriers in treating breast cancer. Hyaluronic acid (HA) is a natural polysaccharide with excellent biocompatibility, biodegradability, and non-immunogenicity and is a significant component of the extracellular matrix. The CD44 receptor of HA is overexpressed in breast cancer cells and can be targeted to breast tumors. Therefore, many researchers have developed nano drug delivery systems (NDDS) based on the CD44 receptor tumor-targeting properties of HA. This review examines the application of HA in NDDSs for breast cancer in recent years. Based on the structural composition of NDDSs, they are divided into HA NDDSs, Modified HA NDDSs, and HA hybrid NDDSs.

Keywords: anticancer drugs; breast cancer; drug delivery system (DDS); hyaluronic acid; nanoparticles.

FIGURE 1

Classification of hyaluronic acid NDDSs in breast cancer.

FIGURE 2

(A) Uncoated NPs, without HA covering (B) HA-NPs at low magnification, (C) HA-NPs at higher magnification. (D) Naproxen release kinetics from HA-coated or uncoated NPs. (E) Percentage of positive cells for c6 and (F) the mean fluorescence intensity per cell (t-student, *p < 0.05, **p < 0.01, ***p < 0.001). In vitro cytotoxicity of HA-NPs in cells with differential expression of CD44. (G) Percentage of cell viability of MCF-7 cells (high expression of CD44) after 72 h of treatment with different concentrations. (H) Cell viability experiments in HepG2 cells (low expression of CD44) using the same NPs. Cell viability assays by Alamar Blue in MCF-7, RAW264.7 and HUVEC cells treated with S-HA-NPs. (I) Percentage of viable MCF-7 cells relative to control (culture media of MCF-7 cells) after 72 h of treatment with different concentrations. (J) Percentage of cell viability in respect to controls (cells treated with culture media) after 72 h of treatment with different concentrations of S-HA-NPs(Statistical analysis was performed by one-way ANOVA test with *p < 0.01, **p < 0.05 and ***p < 0.001). ELISA quantification of (K) PGE2 and (L)* VEGF released by MCF-7 cells after 72 h of treatment (Statistical analysis was performed by one-way ANOVA test with **p < 0.05). Wound healing assay in S-HA-NPs or free NAP treated MCF-7 cells. Effect of different concentrations of S-HA-NPs or free NAP on MCF-7 migration in vitro: (M) Inverted microscope images (20-fold magnification) of the wound at the beginning of the assay (0 h) and 24 h post-scratching and (N) Percentage of open wound after 24 h of treatment when compared to the original wound size (Statistical analysis was performed by one-way ANOVA with *p < 0.01 and **p < 0.05). Reproduced with permission from ref (Liu et al., 2018a). CC BY 4.0. Copyright 2021 The Authors.

FIGURE 3

In vitro cellular uptake and intracellular DOX release in 3T3 and 4T1 cells. (A) Confocal laser scattering microscopy and (B) FCM analysis were performed on 3T3 and 4T1 cells following HA pretreatment, and treatment with free DOX + CDDP and HA-DOX-CDDP. In vitro multicellular spheroids in 3D suspension cultures. (C) Confocal laser scattering microscopy of 4T1 and 3T3 cell spheroids treated with HA, DOX + CDDP, and HA-DOX-CDDP for 24 h. (D) Colony volume and (E) fluorescence density analyses of 4T1 and 3T3 cell spheroids treated with HA, DOX + CDDP, and HA-DOX-CDDP for 24 h (*p < 0.05 compared with the DOX + CDDP group). In vivo DOX biodistribution. (F) Ex vivo fluorescence images of isolated organs and tumors at 6 or 12 h post-injection. (G) Semi-quantitative analysis of the mean fluorescence intensity in isolated organs and tumors at 6 or 12 h post-injection. Data are presented as the mean ± SD (n = 3) (*p < 0.05 compared with the DOX + CDDP group). Reproduced with permission from ref (Owen et al., 2012). CC BY 4.0. Copyright 2020 The Authors.

FIGURE 4

In vivo safety and antitumor efficacies. (A) Tumor volumes and (B) body weights of 4T1-xenografted mice after treatment with NS as the control, DOX + CDDP, or HA-DOX-CDDP. Red arrows showed the tail-veil injection time. (C) Organ coefficients of isolated organs in NS, DOX + CDDP, or HA-DOX-CDDP treated groups (*p < 0.05, #p < 0.001). Histopathology and immunofluorescence analyses. (D) Histopathological (H&E) analyses and (E) necrotic areas in H&E-stained tumor sections from 4T1-xenografted mice following treatment with NS as the control, DOX + CDDP, or HA-DOX-CDDP. Red arrows indicated the necrotic area. (F) Immunohistochemical (PARP and survivin) analyses of tumor tissue sections following treatment with NS as the control, DOX + CDDP, or HA-DOX-CDDP. (G) Relative optical densities of tumor sections showing PARP immunofluorescence. (H) Relative optical densities of tumor sections showing survivin immunofluorescence (Data are presented as the mean ± SD (n = 5). *p < 0.05, **p < 0.01). Reproduced with permission from ref (Owen et al., 2012). CC BY 4.0. Copyright 2020 The Authors.

FIGURE 5

(A) Schematic illustration of the formation and functions of P@CH nanoparticles. (B) NIR thermal imaging before and after NIR irradiation on PBS, PPy NPs, and P@CH NPs plus laser irradiation. (C) Monitoring the increase of temperature with prolonged irradiation time over a series of concentrations of PPy NPs, and P@CH NPs (n = 3). (D) Scheme showing the experimental design to evaluate the therapeutic effect. (E) Infrared thermal images of the tumor-bearing mice at 24 h post-injection of PBS, PPy NPs, and P@CH NPs via the tail vein, before and after NIR laser light irradiation (1.5 W/cm2, 10 min), and the increase of temperature at tumor sites at different time points. (F) Tumor volume measurement for different treatment groups. (G) Quantification of the BLI signal of tumors after different treatments. (H) The body weight of 4T1 tumor-bearing mice with different treatments was measured. (I) Survival analysis of orthotopic 4T1 breast cancer mice with different treatments (n = 5) (**p < 0.01). Immune-memory effects induced by P@CH-L-IT combination treatment. Reproduced with permission from ref (Chen et al., 2019). CC BY 4.0. Copyright 2019 The Authors.

FIGURE 6

(A) Scheme showing the rechallenge experimental design to evaluate the immune-memory effect. (B) The dynamic BLI images of rechallenged mice from the P@CH-L and P@CH-L-IT groups (n = 5) for 65 continuous days. (C) Quantification of the BLI signal intensity of rechallenged tumors. (D) Tumor volume measurement of second rechallenged tumors and first tumors in two groups. Chemo-photothermal therapy in combination with immune checkpoint blockade therapy prevented lung metastases. (E) Representative BLI images and white-light pictures showing the lung metastases. (F) The number of lung metastasis nodules of different groups (n = 3). Reproduced with permission from ref (Chen et al., 2019). CC BY 4.0. Copyright 2019 The Authors.