PEG-poly(β-amino ester) (PEG-PBAE), which is an effective pH-responsive copolymer, was mainly synthesized by Michael step polymerization. Thioridazine (Thz), which was reported to selectively eliminate cancer stem cells (CSCs), was loaded into PEG-PBAE micelles (PPM) prepared by self-assembly at low concentrations. The critical micelle concentrations (CMC) of PPM in water were 2.49 μg/mL. The pH-responsive PBAE segment was soluble due to protonated tertiary amine groups when the pH decreased below pH 6.8, but it was insoluble at pH 7.4. The Thz-loaded PEG-PBAE micelle (Thz/PPM) exhibited a spherical shape, and the drug loading was 15.5%. In vitro release of Thz/PPM showed that this pH-sensitivity triggered the rapid release of encapsulated Thz in a weakly acidic environment. The in vitro cytotoxicity and cellular uptake of various formulations at pH 7.4 and 5.5 were evaluated on the mammospheres (MS), which were sorted by MCF-7 human breast cancer cell lines and identified to be a CD44⁺/CD24⁻ phenotype. The results of the cytotoxicity assay showed that blank micelles were nontoxic and Thz/PPM exhibited a similar anti-CSC effect on MS compared to Thz solution. Stronger fluorescence signal of Coumarin-6 (C6) was observed in MS treated by C6-loaded PPM (C6/PPM) at pH 5.5. The tumor inhibition rate and tumor weight of the free DOX and Thz/PPM groups were significantly different from those of the other groups, which free DOX and Thz/PPM effectively suppressed breast tumor growth in vivo. The above experimental results showed that Thz/PPM is an ideal and effective pH-responsive drug delivery carrier to a targeted therapy of CSCs.

Cancer stem-like cells (CSLCs) have been considered to be one of the main problems in tumor treatment owing to high tumorigenicity and chemotherapy resistance. In this study, we synthesized a novel mitochondria-target derivate, triphentlphosphonium-resveratrol (TPP-Res), and simultaneously encapsulated it with doxorubicin (Dox) in pH-sensitive liposomes (PSL (Dox/TPP-Res)), to reverse chemotherapeutic resistance of CSLCs. PSL (Dox/TPP-Res) was approximately 165 nm in size with high encapsulation efficiency for both Dox and TPP-Res. Cytotoxicity assay showed that the optimal synergistic effect was the drug ratio of 1:1 for TPP-Res and Dox. Cellular uptake and intracellular trafficking assay indicated that PSL (Dox/TPP-Res) could release drugs in acidic endosomes, followed by mitochondrial targeting of TPP-Res and nucleus transports for Dox. The mechanisms for reversing the resistance in CSLCs were mainly attributed to a synergistic effect for reduction of mitochondrial membrane potential, activation of caspase cascade reaction, reduction of ATP level and suppression of the Wnt/β-catenin pathway. Further, in vivo assay results demonstrated that the constructed liposomes could efficiently accumulate in the tumor region and possess excellent antineoplastic activity in an orthotopic xenograft tumor model with no evident systemic toxicity. The above experimental results determined that PSL (Dox/TPP-Res) provides a new method for the treatment of heterogenecity tumors.

Even though the reversal of multi-drug resistance (MDR) by numerous nanoparticles has been extensively studied, limited success has been achieved. To overcome this barrier, we report a rationally-designed pH-sensitive micelle, in which doxorubicin (Dox) and resveratrol (Res) were co-loaded. The micelle was based on methoxy poly (ethylene glycol)-poly(d,l-lactide)-poly(l-histidine) (mPEG-PLA-PHis), which integrated passive targeting, endo-lysosomal escape and pH-responsive payloads release. At a physiological pH of 7.4 (slightly alkali), Dox and Res were incorporated into the micelles core using the thin-film hydration method (pH-endoSM/Dox/Res). After cellular uptake, the micelles exhibited an enhanced dissociation in response to the acidic endosomes, triggering the release of Res and Dox. Furthermore, Res was observed to synergistically improve the cytotoxicity of Dox by down-regulating the P-glycoprotein (P-gp) expression, decreasing the membrane potential of the mitochondrial and ATP level, as well as inducing cell apoptosis mediated by mitochondria. The pH-endoSM/Dox/Res showed a prominent ability to decrease the IC₅₀ of Dox by a factor of 17.38 in cell cytotoxicity against the MCF-7/ADR cell line. In vivo distribution demonstrated the excellent tumor-targeting ability of the pH-endoSM/Dox/Res. All results indicated that pH-endoSM/Dox/Res held great potential for the treatment of Dox-resistance breast cancer cells.

Diabetic chronic wounds face multiple intractable healing obstacles including sustained inflammation, severe infection, insufficient angiogenesis and defective collagen deposition. Current dressings fail to simultaneously relieve all these pathological defects. Herein, we constructed a carrier-free binary self-assembled Glycyrrhizic acid monoammonium salt–Berberine (GB) hydrogel composed of berberine and glycyrrhizic acid monoammonium salt, which forms interconnected nanofiber networks via one-pot thermally assisted small-molecule co-assembly without chemical crosslinking or exogenous polymer carriers. Relying on intermolecular non-covalent interactions, this single supramolecular material integrates anti-inflammatory, broad-spectrum antibacterial, pro-angiogenic, and collagen-regenerative multifunctions, which simultaneously ameliorates multiple core pathological obstacles of diabetic wounds within one formulation. We systematically characterized its physicochemical features, biocompatibility, and antibacterial and anti-inflammatory activities, as well as in vivo wound repair performance. This hydrogel formed uniform nanofibrous architectures with favorable viscoelastic properties and pH-dependent sustained release. In vitro assays verified its outstanding biosafety, broad-spectrum bacteriostasis against Escherichia coli and Staphylococcus aureus, and potent inhibitory effects on pro-inflammatory cytokines TNF-α and IL-6, with bacterial inhibition rates reaching ~88% against E. coli and ~70% against S. aureus. In diabetic mouse full-thickness infected wound models, the GB hydrogel simultaneously alleviated local inflammation, accelerated wound closure and facilitated ordered collagen deposition and mature microvessel formation, achieving a 73.10% wound closure rate at day 7 and 58.01% collagen deposition fraction at day 14, thus exhibiting equivalent or superior repair capacity compared with commercial hydrogel dressings. This carrier-free supramolecular system based on natural herbal small molecules provides a safe, convenient, and integrated therapeutic strategy for diabetic infected chronic wounds, with promising clinical translation potential.