Evaluation of keratin biomaterial containing silver nanoparticles as a potential wound dressing in full-thickness skin wound model in diabetic mice
Corresponding Author
Marek Konop
Department of Experimental Physiology and Pathophysiology, Laboratory of Center for Preclinical Research, Medical University of Warsaw, Warsaw, Poland
Department of Neuropeptides, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland
Department of Dermatology, Medical University of Warsaw, Warsaw, Poland
Correspondence
Marek Konop, Department of Experimental Physiology and Pathophysiology, Laboratory of Center for Preclinical Research, Medical University of Warsaw, 3C A. Pawińskiego Street, Warsaw 02-106, Poland.
Email: [email protected]
Search for more papers by this authorJoanna Czuwara
Department of Dermatology, Medical University of Warsaw, Warsaw, Poland
Search for more papers by this authorEwa Kłodzińska
Department of Analytical Chemistry and Instrumental Analysis, Institute of Sport–National Research Institute, Warsaw, Poland
Search for more papers by this authorAnna K. Laskowska
Department of Neuropeptides, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland
Search for more papers by this authorDorota Sulejczak
Department of Experimental Pharmacology, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland
Search for more papers by this authorTatsiana Damps
Department of Neuropeptides, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland
Department of Dermatology, Medical University of Warsaw, Warsaw, Poland
Search for more papers by this authorUrszula Zielenkiewicz
Department of Microbial Biochemistry, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland
Search for more papers by this authorIwona Brzozowska
Department of Microbial Biochemistry, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland
Search for more papers by this authorAntonio Sureda
Research Group in Community Nutrition and Oxidative Stress and CIBEROBN–Physiopathology of Obesity and Nutrition, University of Balearic Islands, Palma, Spain
Search for more papers by this authorTomasz Kowalkowski
Chair of Environmental Chemistry and Bioanalytics, Faculty of Chemistry, Nicolaus Copernicus University, Toruń, Poland
Interdisciplinary Centre of Modern Technology, Nicolaus Copernicus University, Toruń, Poland
Search for more papers by this authorRobert A. Schwartz
Department of Dermatology, Rutgers New Jersey Medical School, Newark, NJ
Search for more papers by this authorLidia Rudnicka
Department of Neuropeptides, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland
Department of Dermatology, Medical University of Warsaw, Warsaw, Poland
Search for more papers by this authorCorresponding Author
Marek Konop
Department of Experimental Physiology and Pathophysiology, Laboratory of Center for Preclinical Research, Medical University of Warsaw, Warsaw, Poland
Department of Neuropeptides, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland
Department of Dermatology, Medical University of Warsaw, Warsaw, Poland
Correspondence
Marek Konop, Department of Experimental Physiology and Pathophysiology, Laboratory of Center for Preclinical Research, Medical University of Warsaw, 3C A. Pawińskiego Street, Warsaw 02-106, Poland.
Email: [email protected]
Search for more papers by this authorJoanna Czuwara
Department of Dermatology, Medical University of Warsaw, Warsaw, Poland
Search for more papers by this authorEwa Kłodzińska
Department of Analytical Chemistry and Instrumental Analysis, Institute of Sport–National Research Institute, Warsaw, Poland
Search for more papers by this authorAnna K. Laskowska
Department of Neuropeptides, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland
Search for more papers by this authorDorota Sulejczak
Department of Experimental Pharmacology, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland
Search for more papers by this authorTatsiana Damps
Department of Neuropeptides, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland
Department of Dermatology, Medical University of Warsaw, Warsaw, Poland
Search for more papers by this authorUrszula Zielenkiewicz
Department of Microbial Biochemistry, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland
Search for more papers by this authorIwona Brzozowska
Department of Microbial Biochemistry, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland
Search for more papers by this authorAntonio Sureda
Research Group in Community Nutrition and Oxidative Stress and CIBEROBN–Physiopathology of Obesity and Nutrition, University of Balearic Islands, Palma, Spain
Search for more papers by this authorTomasz Kowalkowski
Chair of Environmental Chemistry and Bioanalytics, Faculty of Chemistry, Nicolaus Copernicus University, Toruń, Poland
Interdisciplinary Centre of Modern Technology, Nicolaus Copernicus University, Toruń, Poland
Search for more papers by this authorRobert A. Schwartz
Department of Dermatology, Rutgers New Jersey Medical School, Newark, NJ
Search for more papers by this authorLidia Rudnicka
Department of Neuropeptides, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland
Department of Dermatology, Medical University of Warsaw, Warsaw, Poland
Search for more papers by this authorAbstract
Keratin is a cytoskeletal scaffolding protein essential for wound healing and tissue recovery. The aim of the study was to evaluate the potential role of insoluble fur keratin-derived powder containing silver nanoparticles (FKDP-AgNP) in the allogenic full-thickness surgical skin wound model in diabetic mice. The scanning electron microscopy image evidenced that the keratin surface is covered by a single layer of silver nanoparticles. Data obtained from dynamic light scattering and micellar electrokinetic chromatography showed three fractions of silver nanoparticles with an average diameter of 130, 22.5, and 5 nm. Microbiologic results revealed that the designed insoluble FKDP-AgNP dressing to some extent inhibit the growth of Escherichia coli and Staphylococcus aureus. In vitro assays showed that the FKDP-AgNP dressing did not inhibit fibroblast growth or induce hemolysis. In vivo studies using a diabetic mice model confirmed biocompatible properties of the insoluble keratin dressings. FKDP-AgNP significantly accelerated wound closure and epithelization at Days 5 and 8 (p < .05) when compared with controls. Histological examination of the inflammatory response documented that FKDP-AgNP-treated wounds contained predominantly macrophages, whereas their untreated variants showed mixed cell infiltrates rich in neutrophils. Wound inflammatory response based on macrophages favors tissue remodeling and healing. In conclusion, the investigated FKDP-AgNP dressing consisting of an insoluble fraction of keratin, which is biocompatible, significantly accelerated wound healing in a diabetic mouse model.
CONFLICT OF INTEREST
The authors report no conflicts of interest in this work.
Supporting Information
Filename | Description |
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term2998-sup-0001-Supplementary Materials 31.12.19 do pdf.pdfPDF document, 1.2 MB |
FIGURE S1. Mice during the surgical procedure. Left side – control wound, right side – FKDP+AgNP treated wound. FIGURE S2. Electropherogram obtained for E. coli and S. aureus before treatment with AgNP. FIGURE S3a. Antimicrobial examination of S. aureus, E. coli and B. subtilis after treatment with FKDP, FKDP+AgNP, and control dressing. TABLE 1s. Microbiology results obtained for E. coli and S. aureus. FIGURE S3b. E. coli and S. aureus stained with the LIVE/DEAD bacterial viability kit. Representative micrographs of bacterial cultures after 0, 2h and 24h of FKDP+AgNP or AgNP addition. Live bacteria fluoresce green, dead bacteria fluoresce red. Magnification 1000x. FIGURE S4. a) Cell viability after treatment with AgNP loaded keratin dressing, b) Cell viability after treatment with keratin dressing without AgNP, c) Viability of NIH/3T3 cells after 48 h incubation conditioned medium from FKDP and FKDP-AgNP, d) Effect of silver nanoparticles on murine fibroblasts viability after 24 and 72 h treatment, e) Effect of silver nanoparticles on human red blood cells hemolysis. FIGURE S5. NIH/3T3 and FibStz stained with fibronectin and β-tubulin. FIGURE S6. Wound healing scratch assay after AgNP treatment in different concentrations after 0; 24 and 36 h. Control wells did not contain AgNP. FIGURE S7. Microscopic view of control and FKDP-AgNP-treated wounds during the healing process. H&E. white arrow – neutrophils, black arrow – macrophages, green arrow – multinucleated giant cells, black & white arrow – lymphocytes; magnification 100x. FIGURE S8. a) Tissue biopsy derived from the wound stroma and the wound bed (b) from FKDP-AgNP-treated and non-treated wounds in the process of wound healing, magnification 100x. |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
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