In vitro and in vivo evaluation of implantable bacterial-killing coatings based on host defense peptides and their synthetic mimics

doi:10.1016/j.jmst.2021.02.047

Abstract

Abstract:

Contact-killing antimicrobial coatings based on host defense peptides (HDPs) and their synthetic mimics have shown potential as powerful tools to combat implant-associated infections. Covalent modification of the antimicrobial surface has been utilized to prevent early-stage microbial infections owing to the less drug-leaching possibility that is beneficial to human health and the natural environment. Although considerable progress has been achieved in preparing contact-killing antimicrobial surfaces, discussions focusing on the in vitro and in vivo evaluations of these surfaces are limited. In this review, we summarized the established in vitro methods to simulate the practical interaction of microbes with the surrounding biological environment and the reported in vivo studies at different implant sites. We suggested that the in vivo specific site infection model is essential to gain a comprehensive understanding of these antimicrobial coatings in the preclinical stage, which can be established based on investigations performed using various in vitro assays and conventional non-specific site infection models. Overall, these precedent studies focusing on bacterial contact-killing coatings modified with HDPs and HDP mimics can be considered as critical to assess the surface antibacterial ability and to guide the future developments and applications of antimicrobial surfaces.

Key words: Contact-killing, Antimicrobial surface, HDPs, HDP mimic, Covalent binding

Yuxin Qian, Shuai Deng, Xue Wu, Yunrui She, Runhui Liu, Haodong Lin. In vitro and in vivo evaluation of implantable bacterial-killing coatings based on host defense peptides and their synthetic mimics[J]. J. Mater. Sci. Technol., 2021, 91: 90-104.

Figures/Tables 8

Fig. 1. Development and application of antimicrobial coatings. (A) Different site-specific implant infections in the human body; (Reprinted with permission from Ref. [118]; Copyright (2012), médecine/sciences.) (B) Schematic illustration of extracting an natural HDP from Xenopus laevis and the design of an HDP-mimicking polymer (Reprinted with permission from Ref. [29]; Copyright (2002), National Academy of Sciences.); (C) Statistics of annual journal articles focusing on antimicrobial coatings published from 2000 to 2019 (Archived from Pubmed, keyword: antimicrobial coating); (D) A brief summary of the in vitro and in vivo methods used to evaluate antimicrobial coatings.

Fig. 2. Schematic illustration of several conventional grafting methods for obtaining natural HDP and HDP mimic-modified surfaces.

Fig. 3. Contact-killing antimicrobial surfaces. (A) Thiol-terminated β-peptide polymers were immobilized on the Au surface through “Au-S” bonding (cationic subunits: DM for dimethyl, and lipophilic subunits: CH for cyclohexyl); (Reprinted with permission from Ref. [57]; Copyright (2018), American Chemical Society.) (B) Thiol-terminated β-peptide polymers were immobilized on biomedical substrates through a plasma assisted method, **p < 0.01 (lipophilic subunits: Hex for hexyl); (Reprinted with permission from Ref. [58]; Copyright (2019), American Chemical Society.) (C) Peptide CWR11 was tethered on the PDA (PD, polydopamine) coating on polydimethylsiloxane (PDMS) catheters via a simple dip-coating method; (Reprinted with permission from Ref. [77]; Copyright (2015), Elsevier Ltd.) (D) Dual-functional surface containing an antimicrobial polypeptide: poly(Ppep) and an antifouling polysarcosine: poly(Psar) was modified on various PDA-coated (pDA, polydopamine) surfaces via methacrylate-ended antimicrobial polypeptides (MePpep) and antifouling polypeptoids (MePsar) following UV radiation. (Reprinted with permission from Ref. [81]; Copyright (2017), The Royal Society of Chemistry.).

Fig. 4. Overview of well-established methods to evaluate the antimicrobial activity of contact-killing surfaces. JIS, Japanese Industrial Standards.

Fig. 5. Evaluation of an anti-biofilm surface. (A) The anti-biofilm activity of a polyethylene glycol (PEG)-Mal catheter and CysLasio-III catheter where Mal stands for maleimide; (B) Confocal laser scanning microscopy characterization of Enterococcus faecalis biofilm stained using the LIVE/DEAD assay; (Reprinted with permission from Ref. [114]; Copyright (2017), The Royal Society of Chemistry.) (C) Images of biofilm growth inside of the catheters; (D) Total live bacteria on the inner surfaces of the catheters. Data was obtained using a modified 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. (Reprinted with permission from Ref. [115]; Copyright (2016), American Chemical Society.).

Fig. 6. Subcutaneous implantation model of β-peptide polymer-modified TPU. (A) In vivo antibacterial studies using the subcutaneous implantation of MRSA-contaminated TPU; (B) In vivo antimicrobial activities of the β-peptide polymer-modified TPU recorded after 1 day, 3 days, and 11 days of implantation; (C) Histocompatibility analysis obtained after 1 day, 3 days, and 11 days of subcutaneous implantation. Arrows represent neutrophils. (Reprinted with permission from Ref. [58]; Copyright (2019), American Chemical Society). TPU, thermoplastic polyurethane; TPU-P1, TPU modified with 50:50 DM-Hex; CFU, colony forming units.

Fig. 7. Rabbit eye model to evaluate the antimicrobial ability of Mel4-coated contact lenses. (A) Images recorded under diffuse white light and a fluorescent slit lamp using the control and Mel4-coated contact lenses in rabbit eyes were observed at different time points: day 1, day 8, day 15, and day 22; (B) Representative micrographs of histological sections of rabbit eyes at day 22. (Reprinted with permission from Ref. [135]; Copyright (2017), Elsevier Ltd.).

Fig. 8. Bladder lumen model of a peptide-modified polyurethane (PU) catheter. (A) In Vivo Imaging System (IVIS) images of mice bearing either an uncoated or antimicrobial peptide (AMP)-Brush coated PU catheter recorded at day 1 and day 7 following the post-instillation of P. aeruginosa lux into the bladder; (B) Bioluminescence recorded at day 1, 4, and 7 after the post-bacterial-instillation into mice bladder. Data for the untreated and treated mice were measured using the IVIS® Lumina system; (C) The number of surviving P. aeruginosa recovered from the catheter surface in urine; (D) The number of surviving P. aeruginosa recovered from the catheter surface used in the mice model at 7 days after implantation. *p ≤ 0.05, **p ≤ 0.01, and ***p ≤ 0.001. (Reprinted with permission from Ref. [148]; Copyright (2016), Elsevier Ltd.).

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