Eradication of Mature Bacterial Biofilms with Concurrent Improvement in Chronic Wound Healing Using Silver Nanoparticle Hydrogel Treatment
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Preparation and Characterization of the AgNP Hydrogel
2.3. Stability and Release Kinetics of the AgNP Hydrogel
2.4. Bacterial Bioluminescent Strain and Growth
2.5. In Vitro Cytocompatibility
2.6. Hemolytic Activity of the Test Hydrogels
2.7. In Vitro Anti-Biofilm Eradication of AgNP Hydrogel
2.8. In Vivo Mature Biofilm Wound Infection Animal Model and Treatment
2.9. Macroscopic and Microscopic Analysis of Wound Healing
2.10. In Vivo Antibacterial Assessments
2.10.1. CFU Counts
2.10.2. Bacterial Viability (Live/Dead Staining)
2.11. Statistical Analysis
3. Results and Discussion
3.1. Preparation and Characterization of AgNP Hydrogel
3.2. In Vitro Biocompatibility of the AgNP Hydrogel
3.3. Hemocompatibility of the AgNP Hydrogel
3.4. In Vitro Antibiofilm Activity of the AgNP Hydrogel
3.5. In Vivo Anti-Biofilm Effect of the AgNP Hydrogel
3.6. End-Point Tissue Bacterial Biofilm Analysis
3.7. In Vivo Assessment of AgNP Hydrogel Efficacy in Promoting Healing of Infected Wounds
3.8. Safety of the AgNP Hydrogel Treatment
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Koo, H.; Allan, R.N.; Howlin, R.P.; Stoodley, P.; Hall-Stoodley, L. Targeting microbial biofilms: Current and prospective therapeutic strategies. Nat. Rev. Microbiol. 2017, 15, 740–755. [Google Scholar] [CrossRef]
- Frieri, M.; Kumar, K.; Boutin, A. Antibiotic resistance. J. Infect. Public Health 2017, 10, 369–378. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Flemming, H.-C.; Wingender, J.; Szewzyk, U.; Steinberg, P.; Rice, S.A.; Kjelleberg, S. Biofilms: An emergent form of bacterial life. Nat. Rev. Microbiol. 2016, 14, 563–575. [Google Scholar] [CrossRef] [PubMed]
- Kostakioti, M.; Hadjifrangiskou, M.; Hultgren, S.J. Bacterial biofilms: Development, dispersal, and therapeutic strategies in the dawn of the postantibiotic era. Cold Spring Harb. Perspect. Med. 2013, 3, a010306. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Liu, W.; Ou-Yang, W.; Zhang, C.; Wang, Q.; Pan, X.; Huang, P.; Zhang, C.; Li, Y.; Kong, D.; Wang, W. Synthetic Polymeric Antibacterial Hydrogel for Methicillin-Resistant Staphylococcus aureus-Infected Wound Healing: Nanoantimicrobial Self-Assembly, Drug- and Cytokine-Free Strategy. ACS Nano 2020, 14, 12905–12917. [Google Scholar] [CrossRef] [PubMed]
- Gao, Y.; Wang, J.; Chai, M.; Li, X.; Deng, Y.; Jin, Q.; Ji, J. Size and Charge Adaptive Clustered Nanoparticles Targeting the Biofilm Microenvironment for Chronic Lung Infection Management. ACS Nano 2020, 14, 5686–5699. [Google Scholar] [CrossRef]
- Han, G.; Ceilley, R. Chronic Wound Healing: A Review of Current Management and Treatments. Adv. Ther. 2017, 34, 599–610. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Mi, G.; Shi, D.; Wang, M.; Webster, T.J. Reducing Bacterial Infections and Biofilm Formation Using Nanoparticles and Nanostructured Antibacterial Surfaces. Adv. Funct. Mater. 2018, 7, 1800103. [Google Scholar] [CrossRef]
- Haidari, H.; Garg, S.; Vasilev, K.; Kopecki, Z.; Cowin, A.J. Silver-based wound dressings: Current issues and future developments for treating bacterial infections. Wound Pract. Res. 2020, 28, 176–183. [Google Scholar]
- Liu, Y.; Shi, L.; Su, L.; van der Mei, H.C.; Jutte, P.C.; Ren, Y.; Busscher, H.J. Nanotechnology-based antimicrobials and delivery systems for biofilm-infection control. Chem. Soc. Rev. 2019, 48, 428–446. [Google Scholar] [CrossRef]
- Cheng, H.; Li, Y.; Huo, K.; Gao, B.; Xiong, W. Long-lasting in vivo and in vitro antibacterial ability of nanostructured titania coating incorporated with silver nanoparticles. J. Biomed. Mater. Res. Part A 2014, 102, 3488–3499. [Google Scholar] [CrossRef] [PubMed]
- Kalantari, K.; Mostafavi, E.; Afifi, A.M.; Izadiyan, Z.; Jahangirian, H.; Rafiee-Moghaddam, R.; Webster, T.J. Wound dressings functionalized with silver nanoparticles: Promises and pitfalls. Nanoscale 2020, 12, 2268–2291. [Google Scholar] [CrossRef] [PubMed]
- Barbalinardo, M.; Antosova, A.; Gambucci, M.; Bednarikova, Z.; Albonetti, C.; Valle, F.; Sassi, P.; Latterini, L.; Gazova, Z.; Bystrenova, E. Effect of metallic nanoparticles on amyloid fibrils and their influence to neural cell toxicity. Nano Res. 2020, 13, 1081–1089. [Google Scholar] [CrossRef]
- Decataldo, F.; Barbalinardo, M.; Gentili, D.; Tessarolo, M.; Calienni, M.; Cavallini, M.; Fraboni, B. Organic Electrochemical Transistors for Real-Time Monitoring of In Vitro Silver Nanoparticle Toxicity. Adv. Biosys. 2020, 4, 1900204. [Google Scholar] [CrossRef]
- Rao, B.R.; Kotcherlakota, R.; Nethi, S.K.; Puvvada, N.; Sreedhar, B.; Chaudhuri, A.; Patra, C.R. Ag2[Fe(CN)5NO] Nanoparticles Exhibit Antibacterial Activity and Wound Healing Properties. ACS Biomater. Sci. Eng. 2018, 4, 3434–3449. [Google Scholar] [CrossRef]
- Baek, K.; Liang, J.; Lim, W.T.; Zhao, H.; Kim, D.H.; Kong, H. In Situ Assembly of Antifouling/Bacterial Silver Nanoparticle-Hydrogel Composites with Controlled Particle Release and Matrix Softening. ACS Appl. Mater. Interfaces 2015, 7, 15359–15367. [Google Scholar] [CrossRef]
- Dai, T.; Wang, C.; Wang, Y.; Xu, W.; Hu, J.; Cheng, Y. A Nanocomposite Hydrogel with Potent and Broad-Spectrum Antibacterial Activity. ACS Appl. Mater. Interfaces 2018, 10, 15163–15173. [Google Scholar] [CrossRef] [PubMed]
- Urzedo, A.L.; Gonçalves, M.C.; Nascimento, M.H.M.; Lombello, C.B.; Nakazato, G.; Seabra, A.B. Cytotoxicity and Antibacterial Activity of Alginate Hydrogel Containing Nitric Oxide Donor and Silver Nanoparticles for Topical Applications. ACS Biomater. Sci. Eng. 2020, 6, 2117–2134. [Google Scholar] [CrossRef]
- Taheri, S.; Cavallaro, A.; Christo, S.N.; Smith, L.E.; Majewski, P.; Barton, M.; Hayball, J.D.; Vasilev, K. Substrate independent silver nanoparticle based antibacterial coatings. Biomaterials 2014, 35, 4601–4609. [Google Scholar] [CrossRef] [PubMed]
- Haidari, H.; Kopecki, Z.; Sutton, A.T.; Garg, S.; Cowin, A.J.; Vasilev, K. pH-Responsive “Smart” Hydrogel for Controlled Delivery of Silver Nanoparticles to Infected Wounds. Antibiotics 2021, 10, 49. [Google Scholar] [CrossRef] [PubMed]
- GhavamiNejad, A.; Rajan Unnithan, A.; Ramachandra Kurup Sasikala, A.; Samarikhalaj, M.; Thomas, R.G.; Jeong, Y.Y.; Nasseri, S.; Murugesan, P.; Wu, D.; Hee Park, C.; et al. Mussel-Inspired Electrospun Nanofibers Functionalized with Size-Controlled Silver Nanoparticles for Wound Dressing Application. ACS Appl. Mater. Interfaces 2015, 7, 12176–12183. [Google Scholar] [CrossRef] [PubMed]
- Liu, J.; Li, S.; Fang, Y.; Zhu, Z. Boosting antibacterial activity with mesoporous silica nanoparticles supported silver nanoclusters. J. Colloid Interface Sci. 2019, 555, 470–479. [Google Scholar] [CrossRef]
- Zhang, Y.; Sun, P.; Zhang, L.; Wang, Z.; Wang, F.; Dong, K.; Liu, Z.; Ren, J.; Qu, X. Silver-Infused Porphyrinic Metal–Organic Framework: Surface-Adaptive, On-Demand Nanoplatform for Synergistic Bacteria Killing and Wound Disinfection. Adv. Funct. Mater. 2019, 29, 1808594. [Google Scholar] [CrossRef]
- Haidari, H.; Bright, R.; Strudwick, X.L.; Garg, S.; Vasilev, K.; Cowin, A.J.; Kopecki, Z. Multifunctional ultrasmall AgNP hydrogel accelerates healing of S. aureus infected wounds. Acta Biomater. 2021, 128, 420–434. [Google Scholar] [CrossRef]
- Gurtner, G.C.; Werner, S.; Barrandon, Y.; Longaker, M.T. Wound repair and regeneration. Nature 2008, 453, 314–321. [Google Scholar] [CrossRef] [PubMed]
- Parani, M.; Lokhande, G.; Singh, A.; Gaharwar, A.K. Engineered Nanomaterials for Infection Control and Healing Acute and Chronic Wounds. ACS Appl. Mater. Interfaces 2016, 8, 10049–10069. [Google Scholar] [CrossRef]
- AshaRani, P.V.; Low Kah Mun, G.; Hande, M.P.; Valiyaveettil, S. Cytotoxicity and Genotoxicity of Silver Nanoparticles in Human Cells. ACS Nano 2009, 3, 279–290. [Google Scholar] [CrossRef]
- Zanette, C.; Pelin, M.; Crosera, M.; Adami, G.; Bovenzi, M.; Larese, F.F.; Florio, C. Silver nanoparticles exert a long-lasting antiproliferative effect on human keratinocyte HaCaT cell line. Toxicol. In Vitro 2011, 25, 1053–1060. [Google Scholar] [CrossRef]
- Jose Ruben, M.; Jose Luis, E.; Alejandra, C.; Katherine, H.; Juan, B.K.; Jose Tapia, R.; Miguel Jose, Y. The bactericidal effect of silver nanoparticles. Nanotechnology 2005, 16, 2346–2353. [Google Scholar]
- Chen, G.; Yu, Y.; Wu, X.; Wang, G.; Ren, J.; Zhao, Y. Bioinspired Multifunctional Hybrid Hydrogel Promotes Wound Healing. Adv. Funct. Mater. 2018, 28, 1–10. [Google Scholar] [CrossRef] [Green Version]
- Haidari, H.; Zhang, Q.; Melville, E.; Kopecki, Z.; Song, Y.; Cowin, A.J.; Garg, S. Development of Topical Delivery Systems for Flightless Neutralizing Antibody. J. Pharm. Sci. 2017, 106, 1795–1804. [Google Scholar] [CrossRef] [PubMed]
- Haidari, H.; Goswami, N.; Bright, R.; Kopecki, Z.; Cowin, A.J.; Garg, S.; Vasilev, K. The interplay between size and valence state on the antibacterial activity of sub-10 nm silver nanoparticles. Nanoscale Adv. 2019, 1, 2365–2371. [Google Scholar] [CrossRef] [Green Version]
- Haidari, H.; Kopecki, Z.; Bright, R.; Cowin, A.J.; Garg, S.; Goswami, N.; Vasilev, K. Ultrasmall AgNP-Impregnated Biocompatible Hydrogel with Highly Effective Biofilm Elimination Properties. ACS Appl. Mater. Interfaces 2020, 12, 41011–41025. [Google Scholar] [CrossRef]
- Shakya, S.; He, Y.; Ren, X.; Guo, T.; Maharjan, A.; Luo, T.; Wang, T.; Dhakhwa, R.; Regmi, B.; Li, H.; et al. Ultrafine Silver Nanoparticles Embedded in Cyclodextrin Metal-Organic Frameworks with GRGDS Functionalization to Promote Antibacterial and Wound Healing Application. Small 2019, 15, 1–13. [Google Scholar] [CrossRef]
- Liang, Y.; Zhao, X.; Hu, T.; Han, Y.; Guo, B. Mussel-inspired, antibacterial, conductive, antioxidant, injectable composite hydrogel wound dressing to promote the regeneration of infected skin. J. Colloid Interface Sci. 2019, 556, 514–528. [Google Scholar] [CrossRef]
- Ogunniyi, A.D.; Kopecki, Z.; Hickey, E.E.; Khazandi, M.; Peel, E.; Belov, K.; Boileau, A.; Garg, S.; Venter, H.; Chan, W.Y.; et al. Bioluminescent murine models of bacterial sepsis and scald wound infections for antimicrobial efficacy testing. PLoS ONE 2018, 13, e0200195. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Gurjala, A.N.; Geringer, M.R.; Seth, A.K.; Hong, S.J.; Smeltzer, M.S.; Galiano, R.D.; Leung, K.P.; Mustoe, T.A. Development of a novel, highly quantitative in vivo model for the study of biofilm-impaired cutaneous wound healing. Wound Repair Regen. 2011, 19, 400–410. [Google Scholar] [CrossRef] [PubMed]
- Cowin, A.; Adams, D.; Strudwick, X.; Chan, H.; Hooper, J.; Sander, G.; Rayner, T.; Matthaei, K.; Powell, B.; Campbell, H. Flightless I deficiency enhances wound repair by increasing cell migration and proliferation. J. Pathol. 2007, 211, 572–581. [Google Scholar] [CrossRef] [PubMed]
- Yu, Y.; Li, P.; Zhu, C.; Ning, N.; Zhang, S.; Vancso, G.J. Multifunctional and Recyclable Photothermally Responsive Cryogels as Efficient Platforms for Wound Healing. Adv. Funct. Mater. 2019, 29, 1904402. [Google Scholar] [CrossRef]
- Kopecki, Z.; Arkell, R.; Powell, B.C.; Cowin, A.J. Flightless I Regulates Hemidesmosome Formation and Integrin-Mediated Cellular Adhesion and Migration during Wound Repair. J. Investig. Dermatol. 2009, 129, 2031–2045. [Google Scholar] [CrossRef]
- Han, L.; Lu, X.; Liu, K.; Wang, K.; Fang, L.; Weng, L.-T.; Zhang, H.; Tang, Y.; Ren, F.; Zhao, C.; et al. Mussel-Inspired Adhesive and Tough Hydrogel Based on Nanoclay Confined Dopamine Polymerization. ACS Nano 2017, 11, 2561–2574. [Google Scholar] [CrossRef]
- Tenzer, S.; Docter, D.; Rosfa, S.; Wlodarski, A.; Kuharev, J.; Rekik, A.; Knauer, S.K.; Bantz, C.; Nawroth, T.; Bier, C.; et al. Nanoparticle Size Is a Critical Physicochemical Determinant of the Human Blood Plasma Corona: A Comprehensive Quantitative Proteomic Analysis. ACS Nano 2011, 5, 7155–7167. [Google Scholar] [CrossRef]
- Visalakshan, R.M.; MacGregor, M.N.; Sasidharan, S.; Ghazaryan, A.; Mierczynska-Vasilev, A.M.; Morsbach, S.; Mailänder, V.; Landfester, K.; Hayball, J.D.; Vasilev, K. Biomaterial Surface Hydrophobicity-Mediated Serum Protein Adsorption and Immune Responses. ACS Appl. Mater. Interfaces 2019, 11, 27615–27623. [Google Scholar] [CrossRef]
- Song, X.; Zhu, C.; Fan, D.; Mi, Y.; Li, X.; Fu, R.Z.; Duan, Z.; Wang, Y.; Feng, R.R. A Novel Human-Like Collagen Hydrogel Scaffold with Porous Structure and Sponge-Like Properties. Polymers 2017, 9, 638. [Google Scholar] [CrossRef] [Green Version]
- Lopez Hernandez, H.; Grosskopf, A.K.; Stapleton, L.M.; Agmon, G.; Appel, E.A. Non-Newtonian Polymer–Nanoparticle Hydrogels Enhance Cell Viability during Injection. Macromol. Biosci. 2019, 19, 1800275. [Google Scholar] [CrossRef] [Green Version]
- Sun Han Chang, R.; Lee, J.C.-W.; Pedron, S.; Harley, B.A.C.; Rogers, S.A. Rheological Analysis of the Gelation Kinetics of an Enzyme Cross-linked PEG Hydrogel. Biomacromolecules 2019, 20, 2198–2206. [Google Scholar] [CrossRef]
- Li, J.; Mooney, D.J. Designing hydrogels for controlled drug delivery. Nat. Rev. Mater. 2016, 1, 16071. [Google Scholar] [CrossRef] [PubMed]
- Feng, Y.; Wang, Q.; He, M.; Zhang, X.; Liu, X.; Zhao, C. Antibiofouling Zwitterionic Gradational Membranes with Moisture Retention Capability and Sustained Antimicrobial Property for Chronic Wound Infection and Skin Regeneration. Biomacromolecules 2019, 20, 3057–3069. [Google Scholar] [CrossRef] [PubMed]
- Kopecki, Z.; Ogunniyi, A.D.; Trott, D.J.; Cowin, A. Fighting chronic wound infection—One model at a time. Wound Pract. Res. 2017, 25, 1–6. [Google Scholar]
- Phillips, P.L.; Wolcott, R.D.; Fletcher, J.; Schultz, G.S. Biofilms Made Easy. Wounds Int. 2010, 2010, 1. [Google Scholar]
- Wu, J.; Li, F.; Hu, X.; Lu, J.; Sun, X.; Gao, J.; Ling, D. Responsive Assembly of Silver Nanoclusters with a Biofilm Locally Amplified Bactericidal Effect to Enhance Treatments against Multi-Drug-Resistant Bacterial Infections. ACS Cent. Sci. 2019, 5, 1366–1376. [Google Scholar] [CrossRef] [Green Version]
- Le Ouay, B.; Stellacci, F. Antibacterial activity of silver nanoparticles: A surface science insight. Nano Today 2015, 10, 339–354. [Google Scholar] [CrossRef] [Green Version]
- Patel, K.K.; Surekha, D.B.; Tripathi, M.; Anjum, M.M.; Muthu, M.S.; Tilak, R.; Agrawal, A.K.; Singh, S. Antibiofilm Potential of Silver Sulfadiazine-Loaded Nanoparticle Formulations: A Study on the Effect of DNase-I on Microbial Biofilm and Wound Healing Activity. Mol. Pharm. 2019, 16, 3916–3925. [Google Scholar] [CrossRef] [PubMed]
- Ferry, J.L.; Craig, P.; Hexel, C.; Sisco, P.; Frey, R.; Pennington, P.L.; Fulton, M.H.; Scott, I.G.; Decho, A.W.; Kashiwada, S.; et al. Transfer of gold nanoparticles from the water column to the estuarine food web. Nat. Nanotechnol. 2009, 4, 441–444. [Google Scholar] [CrossRef]
- Gentile, P.; Sterodimas, A.; Pizzicannella, J.; Dionisi, L.; De Fazio, D.; Calabrese, C.; Garcovich, S. Systematic Review: Allogenic Use of Stromal Vascular Fraction (SVF) and Decellularized Extracellular Matrices (ECM) as Advanced Therapy Medicinal Products (ATMP) in Tissue Regeneration. Int. J. Mol. Sci. 2020, 21, 4982. [Google Scholar] [CrossRef] [PubMed]
- Gentile, P.; Garcovich, S. Concise Review: Adipose-Derived Stem Cells (ASCs) and Adipocyte-Secreted Exosomal microRNA (A-SE-miR) Modulate Cancer Growth and promote Wound Repair. J. Clin. Med. 2019, 8, 855. [Google Scholar] [CrossRef] [Green Version]
- Abdal Dayem, A.; Lee, S.B.; Cho, S.G. The Impact of Metallic Nanoparticles on Stem Cell Proliferation and Differentiation. Nanomaterials 2018, 8, 761. [Google Scholar] [CrossRef] [Green Version]
- De Angelis, B.; D’Autilio, M.F.L.M.; Orlandi, F.; Pepe, G.; Garcovich, S.; Scioli, M.G.; Orlandi, F.; Cervelli, V.; Gentile, P. Wound Healing: In Vitro and In Vivo Evaluation of a Bio-Functionalized Scaffold Based on Hyaluronic Acid and Platelet-Rich Plasma in Chronic Ulcers. J. Clin. Med. 2019, 8, 855. [Google Scholar] [CrossRef] [Green Version]
- Zhang, H.; Sun, X.; Wang, J.; Zhang, Y.; Dong, M.; Bu, T.; Li, L.; Liu, Y.; Wang, L. Multifunctional Injectable Hydrogel Dressings for Effectively Accelerating Wound Healing: Enhancing Biomineralization Strategy. Adv. Funct. Mater. 2021, 31, 2100093. [Google Scholar] [CrossRef]
- Huang, W.-C.; Ying, R.; Wang, W.; Guo, Y.; He, Y.; Mo, X.; Xue, C.; Mao, X. A Macroporous Hydrogel Dressing with Enhanced Antibacterial and Anti-Inflammatory Capabilities for Accelerated Wound Healing. Adv. Funct. Mater. 2020, 30, 1–11. [Google Scholar] [CrossRef]
- Gentile, P.; Garcovich, S. Systematic Review: Adipose-Derived Mesenchymal Stem Cells, Platelet-Rich Plasma and Biomaterials as New Regenerative Strategies in Chronic Skin Wounds and Soft Tissue Defects. Int. J. Mol. Sci. 2021, 22, 1538. [Google Scholar] [CrossRef] [PubMed]
- Kopecki, Z. Development of next-generation antimicrobial hydrogel dressing to combat burn wound infection. Biosci Rep. 2021, 41, BSR20203404. [Google Scholar] [CrossRef] [PubMed]
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Haidari, H.; Bright, R.; Garg, S.; Vasilev, K.; Cowin, A.J.; Kopecki, Z. Eradication of Mature Bacterial Biofilms with Concurrent Improvement in Chronic Wound Healing Using Silver Nanoparticle Hydrogel Treatment. Biomedicines 2021, 9, 1182. https://doi.org/10.3390/biomedicines9091182
Haidari H, Bright R, Garg S, Vasilev K, Cowin AJ, Kopecki Z. Eradication of Mature Bacterial Biofilms with Concurrent Improvement in Chronic Wound Healing Using Silver Nanoparticle Hydrogel Treatment. Biomedicines. 2021; 9(9):1182. https://doi.org/10.3390/biomedicines9091182
Chicago/Turabian StyleHaidari, Hanif, Richard Bright, Sanjay Garg, Krasimir Vasilev, Allison J. Cowin, and Zlatko Kopecki. 2021. "Eradication of Mature Bacterial Biofilms with Concurrent Improvement in Chronic Wound Healing Using Silver Nanoparticle Hydrogel Treatment" Biomedicines 9, no. 9: 1182. https://doi.org/10.3390/biomedicines9091182