Inhalable Biomineralized Liposomes for Cyclic Ca2+-Burst-Centered Endoplasmic Reticulum Stress Enhanced Lung Cancer Ferroptosis Therapy.

Lung cancer with the highest mortality poses a great threat to human health. Ferroptosis therapy has recently been raised as a promising strategy for lung cancer treatment by boosting the reactive species (ROS) production and lipid peroxidation (LPO) accumulation intracellularly. However, the insufficient intracellular ROS level and the unsatisfactory drug accumulation in lung cancer lesions hamper the efficacy of ferroptosis therapy. Here, an inhalable biomineralized liposome LDM co-loaded with dihydroartemisinin (DHA) and pH-responsive calcium phosphate (CaP) was constructed as a ferroptosis nanoinducer for achieving Ca2+-burst-centered endoplasmic reticulum (ER) stress enhanced lung cancer ferroptosis therapy. Equipped with excellent nebulization properties, about 6.80-fold higher lung lesions drug accumulation than intravenous injection made the proposed inhalable LDM an ideal nanoplatform for lung cancer treatment. The Fenton-like reaction mediated by DHA with peroxide bridge structure could contribute to intracellular ROS production and induce ferroptosis. Assisted by DHA-mediated sarco-/endoplasmic reticulum calcium ATPase (SERCA) inhibition, the initial Ca2+ burst caused by CaP shell degradation triggered the Ca2+-mediated intense ER stress and subsequently induced mitochondria dysfunction to further boost ROS accumulation, which strengthens ferroptosis. The second Ca2+ burst occurred as a result of Ca2+ influx through ferroptotic pores on cell membranes, thus sequentially constructing the lethal "Ca2+ burst-ER stress-ferroptosis" cycle. Consequently, the Ca2+-burst-centered ER stress enhanced ferroptosis process was confirmed as a cell swelling and cell membrane disruption process driven by notable intracellular ROS and LPO accumulation. The proposed LDM showed an encouraging lung retention property and extraordinary antitumor ability in an orthotropic lung tumor murine model. In conclusion, the constructed ferroptosis nanoinducer could be a potential tailored nanoplatform for nebulization-based pulmonary delivery and underscore the application of Ca2+-burst-centered ER stress enhanced lung cancer ferroptosis therapy.

Two different protein corona formation modes on Soluplus® nanomicelles.

Soluplus® nanomicelles have been widely reported in biomedical field for their excellent drug loading capacity and solubility enhancement ability. However, when administrated in vivo, the protein corona will be formed on Soluplus® nanomicelles, significantly affecting their drug delivery performance. Up to now, few studies examined the protein corona formation process and its impact factors of Soluplus® nanomicelles. The multiple proteins in biofluids may form protein corona in different modes due to their diversified properties. In this study, Bovine serum albumin (BSA), Lysozyme (Lyso) and Bovine hemoglobin (BHb) were chosen as model proteins to investigate the protein corona formation process of Soluplus® nanomicelles. By analyzing the polarity of the protein amino acid residues distributing microenvironments, the results showed that there were two different protein corona formation modes, i.e., surface adsorption and insertion, which were determined by the hydrophilicity of proteins. The hydrophobic BHb followed the insertion mode while hydrophilic BSA and Lyso followed the surface adsorption mode. Ultimately, upon protein corona formation, the size and surface chemistry of nanomicelles was significantly affected. We believe this study will provide a new research paradigm to the design and application of Soluplus® nanomicelles.

Inhalable Biomimetic Protein Corona-Mediated Nanoreactor for Self-Amplified Lung Adenocarcinoma Ferroptosis Therapy.

Ferroptosis therapy by catalyzing the Fenton reaction has emerged as a promising tumor elimination strategy for lung adenocarcinoma (ADC). However, the unsatisfactory Fenton reaction efficiency, strong intracellular antioxidant system, and insufficient lung drug accumulation limits the ferroptosis therapeutic effect. To address these issues, an inhalable nanoreactor was proposed by spontaneously adsorbing biomimetic protein corona (PC) composed of matrix metalloproteinase 2 responsive gelatin and glutamate (Glu) on the surface of cationic nanostructured lipid carriers (NLC) core loaded with ferrocene (Fc) and fluvastatin. The prepared Fc-NLC(F)@PC could be nebulized into lung lesions with 2.6 times higher drug accumulation and boost lipid peroxide production by 3.2 times to enhance ferroptosis therapy. Mechanically, fluvastatin was proved to inhibit monocarboxylic acid transporter 4 mediated lactate efflux, inducing tumor acidosis to boost Fc-catalyzing reactive oxygen species production, while the extracellular elevating Glu concentration was found to inhibit xCT (system Xc-) functions and further collapse the tumor antioxidant system by glutathione synthesis suppression. Mitochondrial dysfunction and cell membrane damage were involved in the nanoreactor-driven ferroptotic cell death process. The enhanced antitumor effects by combination of tumor acidosis and antioxidant system collapse were confirmed in an orthotopic lung ADC tumor model. Overall, the proposed nanoreactor highlights the pulmonary delivery approach for local lung ADC treatment and underscores the great potential of ferroptosis therapy.

Pulmonary delivery nanomedicines towards circumventing physiological barriers: Strategies and characterization approaches.

Pulmonary delivery of nanomedicines is very promising in lung local disease treatments whereas several physiological barriers limit its application via the interaction with inhaled nanomedicines, namely bio-nano interactions. These bio-nano interactions may affect the pulmonary fate of nanomedicines and impede the distribution of nanomedicines in its targeted region, and subsequently undermine the therapeutic efficacy. Pulmonary diseases are under worse scenarios as the altered physiological barriers generally induce stronger bio-nano interactions. To mitigate the bio-nano interactions and regulate the pulmonary fate of nanomedicines, a number of manipulating strategies were established based on size control, surface modification, charge tuning and co-delivery of mucolytic agents. Visualized and non-visualized characterizations can be employed to validate the robustness of the proposed strategies. This review provides a guiding overview of the physiological barriers affecting the in vivo fate of inhaled nanomedicines, the manipulating strategies, and the validation methods, which will assist with the rational design and application of pulmonary nanomedicine.

Bibliometric landscape of the researches on protein corona of nanoparticles.

Unclear biological fate hampers the clinical translation of nanoparticles for biomedical uses. In recent years, it is documented that the formation of protein corona upon nanoparticles is a critical factor leading to the ambiguous biological fate. Efforts have been made to explore the protein corona forming behaviors on nanoparticles, and rearrangement of the relevant studies will help to understand the current trend of such a topic. In this work, the publications about protein corona of nanoparticles in Science Citation Index Expanded database of Web of Science from 2007 to 2020 (1417 in total) were analyzed in detail, and the bibliometrics landscape of them was showcased. The basic bibliometrics characteristics were summarized to provide an overall understanding. Citation analysis was performed to scrutinize the peer interests of these papers. The research hotspots in the field were evaluated, based on which some feasible topics for future studies were proposed. In general, the results demonstrated that protein corona of nanoparticles was a prospective research area, and had attracted global research interests. It was believed that this work could comprehensively highlight the bibliometrics landscape, inspire further exploitation on protein corona of nanoparticles, and ultimately promote the clinical translation of nanoparticles.

Major difference in particle size, minor difference in release profile: a case study of solid lipid nanoparticles.

Solid lipid nanoparticles (SLN) have been widely used in a variety of drug delivery routes, which have the outstanding advantage of controlled drug release. The release of SLN is dominated by many factors, among which the particle size of SLN is a critical one. The aim of this project was to explore the relationship between drug release profile and particle size of SLN. SLN were synthesized via the hot high-pressure homogenization (HPH) method, budesonide (BUD) was used as the model drug, and BUD-SLN1-BUD-SLN4 with increasing particle size was obtained, i.e. 120, 240, 360, and 480 nm. The prepared SLN has good encapsulation efficiency, drug loading capacity, and stability. In vitro release behavior studies showed that the cumulative release of BUD-SLN in Tris-Maleate (Tris-M) media was negligible, while that in Tris-M plus pancreatin media or Tris-M-ethanol media obeyed Ritger-Peppas model or first-order kinetic model, respectively. Noticeably, the release behavior of SLN was to some extent related to the average particle size of SLN, but the correlation was insignificant when the intersection degree of particle size distribution was great. This study provides a new idea for the understanding of in vitro release of SLN and has a certain referencing value for the research and development of novel nanomedicines.

Stability Evaluation of Lyotropic Liquid Crystalline Precursor for the Co-delivery of Chlorhexidine and Silver Nanoparticles.

Sealing the therapeutic agents in the root canal is considered to be an essential step in root canal therapy. The lyotropic liquid crystalline precursor (LLCP) incorporated with chlorhexidine (CHX) and silver nanoparticles (Ag-NPs) has been confirmed as a promising candidate for root canal therapy in the previous study. Importantly, the stability of the LLCP system was a significant determinant for its therapeutic effect and further application. The objective of this study was to comprehensively investigate the stability of the LLCP incorporated with CHX and Ag-NPs. The oil-water partition coefficient of CHX and Ag-NPs was measured. The water absorption and the physical stability of drug-loaded LLCP solution were studied. Stability under high temperature, high humidity, and strong light irradiation was also investigated. The results demonstrated that CHX and Ag-NPs could be entrapped in the water channel of LLCP, indicating the low tendency of drugs leakage. The drug-loaded LLCP was a pseudoplastic fluid and it showed an excellent physical stability with a sedimentation rate of 0.981 and a settling time of 26~28 h. The payload of LLCP was confirmed to weaken the water absorption behavior, which facilitated its transformation to cubic liquid crystal. The stress testing under high temperature, high humidity, and strong light irradiation also manifested that the LLCP was stable when stored under moisture-proof condition. In conclusion, the developed LLCP incorporated with CHX and Ag-NPs was highly stable during storage and qualified for further application.

Impact of particle size and pH on protein corona formation of solid lipid nanoparticles: A proof-of-concept study.

When nanoparticles were introduced into the biological media, the protein corona would be formed, which endowed the nanoparticles with new bio-identities. Thus, controlling protein corona formation is critical to in vivo therapeutic effect. Controlling the particle size is the most feasible method during design, and the influence of media pH which varies with disease condition is quite important. The impact of particle size and pH on bovine serum albumin (BSA) corona formation of solid lipid nanoparticles (SLNs) was studied here. The BSA corona formation of SLNs with increasing particle size (120-480 nm) in pH 6.0 and 7.4 was investigated. Multiple techniques were employed for visualization study, conformational structure study and mechanism study, etc. "BSA corona-caused aggregation" of SLN2‒3 was revealed in pH 6.0 while the dispersed state of SLNs was maintained in pH 7.4, which significantly affected the secondary structure of BSA and cell uptake of SLNs. The main interaction was driven by van der Waals force plus hydrogen bonding in pH 7.4, while by electrostatic attraction in pH 6.0, and size-dependent adsorption was confirmed. This study provides a systematic insight to the understanding of protein corona formation of SLNs.

Nanostructure of DiR-Loaded Solid Lipid Nanoparticles with Potential Bioimaging Functions.

The fluorescence dye-loaded nanoparticles are widely used as bioimaging agents in the field of nanotheranostics. However, the nanoparticles for nanotheranostics usually consist of synthetic materials, such as metal, silica, and organic polymers, which are often biologically incompatible and may arouse toxicity issues. Herein, the potential of near-infrared probe DiR-containing solid lipid nanoparticle suspensions (DiR-SLNS) as the bioimaging agent, which was prepared by lipids and surfactants with excellent biocompatibility, was investigated in this study. The nanostructure of DIR-SLNS system and the distribution of DiR were studied by dissipative particle dynamics (DPD) simulations. The stability of physicochemical properties and fluorescence spectra of DIR-SLNS system were investigated using dynamic laser scattering (DLS), nanoparticle tracking analysis (NTA), and fluorescence spectra. The fluorescence intensity-concentration correlation of DIR-SLNS was also evaluated. As a result, DiR-SLNS demonstrated a "core-shell"-like nanostructure and DiR was mainly distributed in the cetyl palmitate (CP) core rather than the surface of SLNS, which was beneficial to its potential applications in bioimaging. DiR-SLNS exhibited remarkable physicochemical stability as the nanoparticles maintained ~ 90% fluorescence intensity during the 10-day storage time. The correlation between fluorescence intensity and concentration was established and validated using a linear regression model. This study proposed a type of promising candidates in nano-scale with higher safety and fluorescence stability for bioimaging.

Relationship between particle size and lung retention time of intact solid lipid nanoparticle suspensions after pulmonary delivery.

The relationship between the particle size and lung retention time of inhaled nanocarriers was unclear, and this uncertainty hampered the design of nanocarriers for pulmonary delivery. The debate resulted from a lack of knowledge regarding the integrity of the involved nanocarriers. A distinguishable bioimaging probe which could differentiate between integrated and disintegrated nanocarriers by emitting different signals was introduced to address this problem. The aza-BODIPY structured aggregation-caused quenching (ACQ) probes were promising candidates, because they showed intense fluorescence signals in intact nanocarriers while quenched after the decomposition of nanocarriers. This attribute was called an on-off switch. In this paper, ACQ probes were encapsulated into a solid lipid nanoparticle suspension (SLNS) with different particle sizes (120-480 nm), and the relationship between particle size and lung retention time after pulmonary delivery was investigated in BALB/c mice. The results showed that a larger particle size led to a longer lung retention time. By comparing with the results of a non-water-quenching probe, the SLNS systems were found to be mostly intact in the pulmonary region. These findings will serve as a firm basis for the design and development of nanocarriers for pulmonary delivery.