Advanced Healthcare MaterialsVolume 13, Issue 17 2470109 Inside Front CoverFree Access Histatin 5-Inspired Short-Chain Peptides Selectively Combating Pathogenic Fungi with Multifaceted Mechanisms (Adv. Healthcare Mater. 17/2024) Guojun Lu, Guojun LuSearch for more papers by this authorXiaoyan Ju, Xiaoyan JuSearch for more papers by this authorMeng Zhu, Meng ZhuSearch for more papers by this authorJinzhao Ou, Jinzhao OuSearch for more papers by this authorDandan Xu, Dandan XuSearch for more papers by this authorKejia Li, Kejia LiSearch for more papers by this authorWei Jiang, Wei JiangSearch for more papers by this authorChenxiao Wan, Chenxiao WanSearch for more papers by this authorYe Tian, Ye TianSearch for more papers by this authorZhongwei Niu, Zhongwei NiuSearch for more papers by this author Guojun Lu, Guojun LuSearch for more papers by this authorXiaoyan Ju, Xiaoyan JuSearch for more papers by this authorMeng Zhu, Meng ZhuSearch for more papers by this authorJinzhao Ou, Jinzhao OuSearch for more papers by this authorDandan Xu, Dandan XuSearch for more papers by this authorKejia Li, Kejia LiSearch for more papers by this authorWei Jiang, Wei JiangSearch for more papers by this authorChenxiao Wan, Chenxiao WanSearch for more papers by this authorYe Tian, Ye TianSearch for more papers by this authorZhongwei Niu, Zhongwei NiuSearch for more papers by this author First published: 05 July 2024 https://doi.org/10.1002/adhm.202470109AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinkedInRedditWechat Graphical Abstract Selectively Combating Fungi The peptide WHI selectively combats fungi without destroying commensal bacteria. When WHI meets the cell membrane, it changes from the PPII conformation to α-helix conformation and targets the nucleus and mitochondria, leading to fungal death. More details can be found in article 2303755 by Zhongwei Niu and co-workers. Volume13, Issue17July 5, 20242470109 RelatedInformation

Short-chain antifungal peptides (AFPs) inspired by histatin 5 have been designed to address the problem of antifungal drug resistance. These AFPs demonstrate remarkable antifungal activity, with a minimal inhibitory concentration as low as 2 µg mL-1. Notably, these AFPs display a strong preference for targeting fungi rather than bacteria and mammalian cells. This is achieved by binding the histidine-rich domains of the AFPs to the Ssa1/2 proteins in the fungal cell wall, as well as the reduced membrane-disrupting activity due to their low amphiphilicity. These peptides disrupt the nucleus and mitochondria once inside the cells, leading to reactive oxygen species production and cell damage. In a mouse model of vulvovaginal candidiasis, the AFPs demonstrate not only antifungal activity, but also promote the growth of beneficial Lactobacillus spp. This research provides valuable insights for the development of fungus-specific AFPs and offers a promising strategy for the treatment of fungal infectious diseases.

Peptide vaccines induce specific neutralizing antibodies and are effective in disease prevention and treatment. However, peptide antigens have a low immunogenicity and are unstable, requiring efficient vaccine carriers to enhance their immunogenicity. Here, we develop a tobacco mosaic virus (TMV)-based peptide vaccine for transdermal immunization using a tip-loaded dissolving microneedle (MN) patch. TMV is decorated with the model peptide antigen PEP3. The prepared TMV-PEP3 promotes dendritic cell maturation and induces dendritic cells to overexpress MHC II, costimulatory factors, and pro-inflammatory factors. By encapsulation of TMV-PEP3 in the tips of a trehalose MN, TMV-PEP3 can be delivered by MN and significantly promote local immune cell infiltration. In vivo studies show that both subcutaneous injection and MN administration of TMV-PEP3 increase the production of anti-PEP3 IgG antibodies and the harvested serum can induce complement-dependent cytotoxicity. This work provides a promising strategy for constructing efficient and health-care-friendly peptide vaccines.

Osteomyelitis caused by methicillin-resistant Staphylococcus aureus (MRSA) is a challenging and life-threatening disease due to its long duration and deep site of occurrence. Herein, we design an in situ formed antibacterial hydrogel with collagenase-respontive activity for the prevention of MRSA-induced osteomyelitis. This hydrogel is constructed by chemical bonding between a 4-arm polyethylene glycol maleimide (4-Arm-PEG-Mal), an N-terminal maleimidated antimicrobial peptide (Mal-AMP) and a collagenase-cleavable peptide with two thiol groups (HS-VPM-SH). Collagenase-induced hydrogel cleavage confers on-demand delivery of the antimicrobial peptide, thereby enhancing the combating activity against MRSA in an infected environment. The designed hydrogel, namely AMP/VPM/PEG hydrogel, can be formed rapidly in situ under physiological conditions due to the rapid bonding between the maleimide and thiol groups. In the in vivo rat osteomyelitis model, the AMP/VPM/PEG hydrogel can be administered by simply injecting the 4-Arm-PEG-Mal and peptide solutions with a 26G needle, which is a minimally invasive method. In vivo evaluation further demonstrates that AMP/VPM/PEG hydrogel can successfully prevent MRSA-induced osteomyelitis. This work provides a minimally invasive approach for intramedullary delivery of antimicrobial peptides on-demand and may provide a viable strategy for the osteomyelitis prevention in clinic.

The phenotype of tumor-associated macrophages plays an important role in their function of regulating the tumor immune microenvironment. The M1-phenotype macrophages display tumor-killing and immune activating functions. Here we show that the tobacco mosaic virus (TMV), a rod-like plant virus, can polarize macrophages to an M1 phenotype and shape a tumor-suppressive microenvironment. RAW 264.7 cells and bone marrow derived-macrophages (BMDMs) can recognize TMV via Toll-like receptor-4, and then the MAPK and NF-κB signaling pathways are activated, leading to the production of pro-inflammatory factors. Furthermore, the in vivo assessments on a subcutaneous co-injection tumor model show that the TMV-polarized BMDMs shape a tumor-suppressive microenvironment, resulting in remarkable delay of 4T1 tumor growth. Another in vivo assessment on an established tumor model indicates the high tumor-metastasis-inhibiting capacity of TMV-polarized BMDMs. This work suggests a role for this plant virus in macrophage-mediated therapeutic approaches and provides a strategy for tumor immunotherapy.

Surface-associated microbe contamination by Gram-negative bacteria poses a serious problem in medical care. Cationic peptides or polymers are the main materials used for antibacterial surface coating, but the positive charge may lead to blood coagulation. Therefore, exploiting surface coating which is free of positive charge and is effective for Gram-negative bacteria inactivation is in urgent need. In this study, inspired by the affinity between lipopolysaccharides of Gram-negative bacteria and Toll-like receptors of immune cells, we develop a leucine-based tetrapeptide coating strategy for combating Gram-negative bacteria. The obtained surface has excellent bactericidal activity against Gram-negative bacteria like Pseudomonas aeruginosa and Escherichia coli. A 1 mm2 coated glass surface could kill > 9.9 × 104 CFU bacteria in 1 h and has nearly no damage to mammal cells. Moreover, this surface coating strategy could be applied on various surfaces like glass slices, glass capillary cavity and thermoplastic polyurethane slices. And the coated surface could largely mitigate the microbe contamination in an in vivo subcutaneous implantation. This work paves a new way for antibacterial surface-coating which is behaving no positive charge and is of great importance for biomedical devices.

Fungal hyphae deeply invade the cornea in fungal keratitis. The corneal stroma hinders the infiltration of antifungal drugs and reduces their bioavailability. Here, this work reports a peptide conjugate nano-assembly that permeates the stroma and kills the pathogen without irritating the ocular cornea. The hydrophilic surface of the nano-assembly ensures deep permeation into the stroma. When encountering a fungal hyphal cell, the nano-assembly disassembles and exposes the α-helical peptide to destroy the fungal membrane, thus inactivating the pathogen. In a rabbit model of fungal keratitis, the nano-assembly exhibits a better therapeutic effect than commercially available natamycin ophthalmic suspension. Peptide conjugates with a nano-assembled structure and assembly-disassembly behavior could serve as the foundation of a new therapy for fungal keratitis.

Microencapsulated recombinant cells technology is a novel approach to tumors therapy. It is necessary to prepare a plenty of the microcapsules with better cell viability and higher endostatin production in order to bring this technology into the clinic. The in vitro culture and cryopreservation are very important parameters in the preparation of microencapsulated cells. In this work, we studied the effect of the in vitro culture and cryopreservation on microencapsulated recombinant cells growth and endostatin production and the effect of the in vitro culture on the cryopreservation of microencapsulated recombinant cells. The results showed that the time of in vitro culture potently affected microencapsulated recombinant CHO cells growth in vivo, endostatin production and the microcapsule stability. The microcapsule kept intact after 36 days of implantation when the in vitro culture time was under 4 days. The thawed microencapsulated recombinant CHO cells had better cell growth and higher endostatin production after 40 days of cryopreservation when the in vitro culture time was 4 days and 8 days. Therefore, the best in vitro culture time was 4 days according to the results of the in vivo culture and cryopreservation and the cryopreservation did not affect microencapsulated recombinant CHO cells growth in vivo, endostatin production and the microcapsule stability.

The transplantation of microencapsulated recombinant cells is a novel alternative approach for the treatment of tumors through gene therapy, whereas its clinical application was retarded because the technique for large-scale preparation and culture of microencapsulated cells is still immature. Optimization of the preparation and culture conditions is needed to acquire biological microcapsule with high cell viability and protein production. In this study, the effects of different preparation and culture conditions on microencapsulated recombinant CHO cell growth and endostatin production were studied. The results showed that the growth phase of the inoculum cells and the seeding density potently affected the growth and endostatin production of the recombinant CHO cells in the microcapsule. The exponential growth phase of the recombinant CHO cells with a seeding density of 1×106–2×106cells/mL microcapsule favored cell growth and endostatin production. The preparation time was another important factor that affected cell viability; the preparation time should be controlled within 5 h to avoid more damage to cells. There would be some damage to cells in the microencapsulation process, and the in vitro culture of microencapsulated cells was a suitable method to recover the cell viability. The highest viable cell density and endostatin production were acquired when the microcapsule percentage was 5% in the culture of microencapsulated cells.