Ultrathin Clay Nanoparticles-Mediated Mutual Reinforcement of Ferroptosis and Cancer Immunotherapy.

Ferroptosis-triggered immunogenic cell death (ICD) is widely adopted to potentiate the body's antitumor immunity by catalyzing the production of toxic reactive oxygen species (ROS). However, the efficacy of ferroptosis and immunotherapy is greatly restricted by intracellular abundant glutathione (GSH) and immunosuppressive tumor microenvironment (TME). Herein, a facile bottom-up method for solvent-free synthesis of ultrathin manganese (Mn)-based layered double hydroxide nanosheets with high loading efficiency for pro-inflammatory cytokine interferon (IFNγ) (IFNγ/uMn-LDHs) is proposed to mutually reinforce the ferroptosis and systemic immunity. The introduction of manganese ions significantly contributes to GSH depletion and hydroxyl radical generation, which can be further enhanced by IFNγ delivery-induced SLC7A11 downregulation. The ICD effect after cell ferroptosis cooperates with the intrinsic immunomodulatory property of IFNγ/uMn-LDHs to facilitate the maturation of dendritic cells (DCs) and the priming of T cells. IFNγ secretion from activated CD8+ T cells in turn involves cascade immunogenic ferroptosis, thus constructing a closed-loop therapy. Remarkably, a potent abscopal effect is observed in the growth inhibition of both primary and distant tumors. Overall, the ultrathin Mn-based clay nanoplatform provides a simple approach for mutual regulation between ferroptosis and antitumor immune response, overcoming the obstacles of current cancer immunotherapy.

Ultrasmall Self-Cascade AuNP@FeS Nanozyme for H2S-Amplified Ferroptosis Therapy.

Recently, nanozymes with peroxidase (POD)-like activity have shown great promise for ferroptosis-based tumor therapy, which are capable of transforming hydrogen peroxide (H2O2) to highly toxic hydroxyl radicals (•OH). However, the unsatisfactory therapeutic performance of nanozymes due to insufficient endogenous H2O2 and acidity at tumor sites has always been a conundrum. Herein, an ultrasmall gold (Au) @ ferrous sulfide (FeS) cascade nanozyme (AuNP@FeS) with H2S-releasing ability constructed with an Au nanoparticle (AuNP) and an FeS nanoparticle (FeSNP) is designed to increase the H2O2 level and acidity in tumor cells via the collaboration between cascade reactions of AuNP@FeS and the biological effects of released H2S, achieving enhanced •OH generation as well as effective ferroptosis for tumor therapy. The cascade reaction in tumor cells is activated by the glucose oxidase (GOD)-like activity of AuNP in AuNP@FeS to catalyze intratumoral glucose into H2O2 and gluconic acid; meanwhile, the released H2S from AuNP@FeS reduces H2O2 consumption by inhibiting intracellular catalase (CAT) activity and promotes lactic acid accumulation. The two pathways synergistically boost H2O2 and acidity in tumor cells, thus inducing a cascade to generate abundant •OH by catalyzing H2O2 through the POD-like activity of FeS in AuNP@FeS and ultimately causing amplified ferroptosis. In vitro and in vivo experiments demonstrated that AuNP@FeS presents a superior tumor therapeutic effect compared to that of AuNP or FeS alone. This strategy represents a simple but powerful method to amplify ferroptosis with H2S-releasing cascade nanozymes and will pave a new way for the development of tumor therapy.

Strong infiltrative HHC36 antimicrobial peptide/silver nanoparticles-loaded carboxymethyl chitosan/sodium alginate hydrogel for acne vulgaris therapy.

Acne is a common chronic skin inflammatory disease closely related toCutibacterium acnes(C. acnes), which affects the life quality of patients worldwide, especially adolescents and young adults. However, the physical barrier of the skin makes drugs difficult to infiltrate effectively into infected site, causing acne hard to cure and easy to recur. Herein, we developed an antibacterial skin dressing with strong infiltration of antibacterial agents which can co-delivery small-molecular antimicrobial agents through stratum corneum deeply into dermis, achieving high antimicrobial efficacy. The antibacterial dressings were constructed with carboxymethyl chitosan/sodium alginate (CMCS/SA) hydrogel loading with HHC36 (an antimicrobial peptide) and silver nanoparticles (AgNPs) conjugates (Ag-H2/CMCS/SA hydrogel). The released Ag-H2from Ag-H2/CMCS/SA hydrogel can early infiltrate into dermis, co-delivery HHC36 and AgNPs due to the infiltration and targeting of HHC36, presenting the superior antibacterial effect compared to HHC36 or AgNPs alone and killing 100%C. acnesand 100%Staphylococcus epidermidis(S. epidermidis) at a very low concentration of Ag-H2(15µg ml-1A g with 7.1µg ml-1HHC36). Meanwhile, Ag-H2/CMCS/SA hydrogel was biocompatible due to the natural polysaccharides carboxymethyl chitosan and sodium alginate. The HaCaT cells spread well in Ag-H2/CMCS/SA hydrogel. These results indicate that the co-delivery small-molecular antimicrobial agents is a promising strategy and Ag-H2/CMCS/SA hydrogel has a great potential in the therapy of acne.

Antimicrobial Titanium Surface via Click-Immobilization of Peptide and Its in Vitro/Vivo Activity.

The use of antimicrobial peptides (AMPs)-functionalized titanium implants is an efficient method for preventing bacterial infection. However, the attachment of AMPs to the surface of titanium implants remains a challenge. In this study, a "clickable" titanium surface was developed by using a silane coupling agent with an alkynyl group. The antimicrobial titanium implant was then constructed through the reaction between the "clickable" surface and azido-AMPs (PEG-HHC36:N3-PEG12-KRWWKWWRR) via click chemistry of Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC). Such an antimicrobial titanium implant, with an AMP density of 897.4 ± 67.3 ng/cm2 (2.5 ± 0.2 molecules per nm2) on the surface, exhibited good and stable antimicrobial activity, inhibited 90.2% of Staphylococcus aureus and 88.1% of Escherichia coli after 2.5 h of incubation, and even inhibited 69.5% of Staphylococcus aureus after 4 days of degradation. The CCK-8 assay indicated that the antimicrobial titanium implant exhibited negligible cytotoxicity to mouse bone mesenchymal stem cells. In vivo assay illustrated that this implant could kill 78.8% of Staphylococcus aureus after 7 days. This method has great potential for the preparation of antimicrobial titanium implants and the prevention of infections in the clinic.

Temperature-Controlled Reversible Exposure and Hiding of Antimicrobial Peptides on an Implant for Killing Bacteria at Room Temperature and Improving Biocompatibility in Vivo.

Modification of implants by antimicrobial peptides (AMPs) can improve the antimicrobial activity of the implants. However, AMPs have some cytotoxicity in vivo when they are exposed at body temperature. To tackle this challenge, we propose to develop a new approach to generating a smart antimicrobial surface through exposure of AMPs on the surface. A polydopamine film was first formed on the substrates, followed by the conjugation of a temperature-sensitive polymer, poly( N-isopropylacrylamide) (pNIPAM), to the film through atom transfer radical polymerization (ATRP). Then, AMPs were conjugated to the NIPAM on the resultant pNIPAM-modified surface through a click chemistry reaction. Because of the temperature-sensitive property of pNIPAM, the AMPs motif was more exposed to the external environment at room temperature (25 °C) than at body temperature (37 °C), making the surface present a higher antimicrobial activity at room temperature than at body temperature. More importantly, such a smart behavior is accompanied with the increased biocompatibility of the surface at body temperature when compared to the substrates unmodified or modified by AMPs or pNIPAM alone. Our in vivo study further verified that pNIPAM-AMP dual modified bone implants showed increased biocompatibility even when they were challenged with the bacteria at room temperature before implantation. These results indicate that the implants are antibacterial at room temperature and can be safely employed during surgery, resulting in no infection after implantations. Our work represents a new promising strategy to fully explore the antimicrobial property of AMPs, while improving their biocompatibility in vivo. The higher exposure of AMPs at room temperature (the temperature for storing the implants before surgery) will help decrease the risk of bacterial infection, and the lower exposure of AMPs at body temperature (the temperature after the implants are placed into the body by surgery) will improve the biocompatibility of AMPs.

Immobilization of an antimicrobial peptide on silicon surface with stable activity by click chemistry.

Infections associated with biomedical implants and devices pose a serious clinical challenge in hospitals worldwide. Antimicrobial peptides (AMPs) have become a great prospect to inhibit this type of infection due to their broad-spectrum antimicrobial activity and low cytotoxicity. However, it is still a challenge to apply AMPs on the biomaterial surface as the activity of AMPs is sensitive to salt or enzyme. In the present study, we prepared a spacer molecule, poly[2-(methacryloyloxy)ethyl]dimethyl-(3-sulfopropyl)ammonium hydroxide (polySBMA), on a model silicon surface via surface-initiated atom transfer radical polymerization (SI-ATRP). We then modified the antimicrobial peptide HHC36 (KRWWKWWRR) with l-propargylglycine (PraAMP) to improve its salt-tolerant activity and integrated PraAMP onto the spacer molecule using click chemistry. We employed X-ray photoelectron spectroscopy (XPS), contact angle goniometry, and atomic force microscopy (AFM) to confirm the success of the immobilization process. We also characterized the antimicrobial activity and stability of the surface with an antimicrobial assay. The results reveal that the modified surface exhibits good antimicrobial activity to inhibit 98.26% of E. coli, 83.72% of S. aureus, and 81.59% of P. aeruginosa. Furthermore, as compared to the control group without the polySBMA spacer, the modified surface improved its resistance to enzymolysis. An in vitro CCK-8 assay also illustrated that this surface showed negligible cytotoxicity to mouse bone mesenchymal stem cells.

Improvement of Uveal and Capsular Biocompatibility of Hydrophobic Acrylic Intraocular Lens by Surface Grafting with 2-Methacryloyloxyethyl Phosphorylcholine-Methacrylic Acid Copolymer.

Biocompatibility of intraocular lens (IOL) is critical to vision reconstruction after cataract surgery. Foldable hydrophobic acrylic IOL is vulnerable to the adhesion of extracellular matrix proteins and cells, leading to increased incidence of postoperative inflammation and capsule opacification. To increase IOL biocompatibility, we synthesized a hydrophilic copolymer P(MPC-MAA) and grafted the copolymer onto the surface of IOL through air plasma treatment. X-ray photoelectron spectroscopy, atomic force microscopy and static water contact angle were used to characterize chemical changes, topography and hydrophilicity of the IOL surface, respectively. Quartz crystal microbalance with dissipation (QCM-D) showed that P(MPC-MAA) modified IOLs were resistant to protein adsorption. Moreover, P(MPC-MAA) modification inhibited adhesion and proliferation of lens epithelial cells (LECs) in vitro. To analyze uveal and capsular biocompatibility in vivo, we implanted the P(MPC-MAA) modified IOLs into rabbits after phacoemulsification. P(MPC-MAA) modification significantly reduced postoperative inflammation and anterior capsule opacification (ACO), and did not affect posterior capsule opacification (PCO). Collectively, our study suggests that surface modification by P(MPC-MAA) can significantly improve uveal and capsular biocompatibility of hydrophobic acrylic IOL, which could potentially benefit patients with blood-aqueous barrier damage.

Preparation of an antimicrobial surface by direct assembly of antimicrobial peptide with its surface binding activity.

Antimicrobial peptides (AMPs) are a broad prospect for clinical application against bacterial infections of biomaterials. However, a bottleneck exists as there is a lack of simple technology to prepare AMPs on biomaterials with sufficient activity, as the activity of AMP is dependent on the correct orientation on the biomaterial. In the present study, based on the conventional AMP (Tet213: KRWWKWWRRC) and surface binding peptide (SKHKGGKHKGGKHKG), we designed an Anchor-AMP that could be directly assembled onto the surface of the biomaterial and also showed excellent antimicrobial activity. By characterizing the surface using a quartz crystal microbalance with dissipation (QCM-D), contact angle, atom force microscopy (AFM) and X-ray photoelectron spectroscopy (XPS), we found that Anchor-AMP could adsorb onto the titanium surface with a strong affinity. Different from Tet213 peptide, Anchor-AMP exhibits excellent antimicrobial activity on the titanium surface being able to inhibit 95.33% of Escherichia coli and 96.67% of Staphylococcus aureus after 2.5 h. The improved antimicrobial activity is a result of improved orientation of Anchor-AMP on the biomaterial compared to that of the Tet213 peptide. In addition, the antimicrobial activity of Anchor-AMP was active for more than 24 h. The CCK-8 assay illustrated that the modified titanium surface showed negligible cytotoxicity to bone marrow mesenchymal stem cells. The in vivo results showed that it exhibited excellent antimicrobial activity after 5 and 7 days, inhibiting 89.32% and 99.78% of S. aureus, respectively. We also demonstrated that Anchor-AMP could be applied on a variety of surfaces including gold (Au), polymethyl methacrylate (PMMA) and hydroxyapatite (HA) with strong affinity and good antimicrobial activity.

Thermoresponsive Self-Assembled ß-Cyclodextrin-Modified Surface for Blood Purification.

For patients with liver failure, bilirubin (BR) is one of the endogenous toxins in their blood. Although blood purification can remove the bilirubin from the body in clinics, the detoxification system needs to be improved, and the cost needs to be decreased. In the present study, we developed a recyclable model surface that can strongly remove bilirubin. We first prepared adamantane (Ad) on a model gold surface by self-assembly. Then, we integrated the ß-cyclodextrin dimer (CDD) onto the surface with host-guest interactions between one of the CD cavities in the CDD and Ad. We characterized the surface with XPS, static contact angle measurements, and AFM. In addition, we employed QCM-D to characterize the preparation process as well as the detoxification of the surface. We demonstrated that this modified surface could strongly adsorb bilirubin through host-guest interactions between the CD cavities in the CDD and bilirubin and that the detoxification was improved 1.7 times (compared to the surface only with Ad). Interestingly, after characterization with QCM-D, this surface could be recycled due to the thermoresponsive property of the host-guest interaction between the CDD and Ad. After adsorbing the toxin and increasing the temperature to 45 °C, the CDD with bilirubin could be removed from the surface. Then, the refreshed surface with CDD could be prepared again at room temperature. This cycle could be repeated at least 3 times. Additionally, during each cycle, the modified surface exhibited good detoxification to bilirubin. This modified surface also showed strong resistance to plasma proteins, decreasing the adsorption of human serum albumin (HSA) and fibrinogen (Fg). An in vitro platelet adhesion assay showed that the adhesion of the platelets on the modified surface decreased and that the platelets were in an inactivated state. The hemolysis assay showed that this surface exhibited no hemolysis activity in the samples to red blood cells (RBCs). The CCK-8 assay showed that this surface had negligible cytotoxicity to L929 cells. This work has taken advantage of the host-guest self-assembly between ß-CD and BR/Ad for special recognizing adsorption, as well as the thermoresponse of ß-CD-Ad inclusion for recyclable application, and these results demonstrate that this technology has great potential for removing bilirubin and decreasing clinic costs.

Antimicrobial Hyaluronic Acid/Poly(amidoamine) Dendrimer Multilayer on Poly(3-hydroxybutyrate-co-4-hydroxybutyrate) Prepared by a Layer-by-Layer Self-Assembly Method.

In this article, we prepared hyaluronic acid/poly(amidoamine) dendrimer (HA/PAMAM) multilayers on a poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-4HB)] substrate by a layer-by-layer self-assembly method for antimicrobial biomaterials. The results of ζ potential and quartz crystal microbalance with dissipation (QCM-D) showed that HA/PAMAM multilayers could be formed on the substrate layer by layer. We used QCM-D to show that both the HA outer layer and the PAMAM outer layer exhibited good protein-resistant activity to bovine serum albumin and bacterial antiadhesion activity to Escherichia coli. By a live/dead assay and the colony counting method, we found that the PAMAM outer layer could also exhibit bactericidal activity against E. coli, while the HA outer layer had no bactericidal activity. Both the bacterial antiadhesion activity and the bactericidal activity of the samples could be maintained even after storage in phosphate-buffered saline for up to 14 days. An in vitro MTT assay showed that the multilayers had no cytotoxicity to L929 cells, and HA molecules in the multilayers could improve the biocompatibility of the film.