This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
University of Belgrade, Faculty of Technology and Metallurgy, Karnegijeva 4 , Belgrade , Serbia
University of Belgrade, Faculty of Technology and Metallurgy, Karnegijeva 4 , Belgrade , Serbia
University of Belgrade, Faculty of Technology and Metallurgy, Karnegijeva 4 , Belgrade , Serbia
University of Belgrade, Innovation Center of the Faculty of Technology and Metallurgy, Karnegijeva 4 , Belgrade , Serbia
University of Belgrade, Institute for the Application of Nuclear Energy INEP, Banatska 31b , Belgrade , Serbia
,
University of Belgrade, Faculty of Technology and Metallurgy, Karnegijeva 4 , Belgrade , Serbia
University of Belgrade, Innovation Center of the Faculty of Technology and Metallurgy, Karnegijeva 4 , Belgrade , Serbia
Puffballs are a group of macrofungi that produce enclosed, globose fruiting bodies. When young, they have a solid white interior (gleba), which turns into a powdery, brown spore-bearing mass as they mature. These fungi have been used in folk medicine worldwide, primarily as wound dressings. Since fungal β-glucans are known to aid in wound healing, this study utilized the β-glucan-enriched fraction from the pestle puffball (Lycoperdon excipuliforme) to synthesize silver nanoparticles (AgNPs), which were incorporated into alginate-based hydrogels for potential wound treatment. Silver nanoparticles were synthesized via a hydrothermal method using silver nitrate, sodium citrate, and varying concentrations of β-glucan extract as stabilizing agents: 0.4 mg/mL (S1), 0.8 mg/mL (S2), 1.6 mg/mL (S3), and 2.4 mg/mL (S4). Particle size was approximately 30 nm for all samples, with zeta potential ranging from -22.3 to -13, increasing with the extract concentration. A one-year stability study at room temperature showed minimal changes in UV-VIS spectra and particle size distribution, despite relatively low zeta potential, suggesting steric stabilization by β-glucan extract. To assess stability in physiological conditions, samples were diluted in 0.85% NaCl solution, and size distribution and UV-VIS absorption were monitored for seven days. Sample S4 exhibited the highest stability and was selected for further experiments. Its antibacterial activity was tested against common wound-infecting bacteria, Staphylococcus aureus and Pseudomonas aeruginosa, using the broth microdilution method. The minimum inhibitory concentration (MIC) was 35 µg/mL for both strains, with minimum bactericidal concentrations (MBC) of 120 µg/mL and 240 µg/mL, respectively. S4 was then used to prepare an alginate-based hydrogel, and a release study was conducted to assess the product’s potential efficacy. The hydrogel was made by mixing equal volumes of AgNP suspension and 4% sodium alginate solution, with the addition of 4% CaCl₂ solution in a circular mold. The release study, conducted in TRIS buffer (50 mM) with 0.85% NaCl at varying pH levels over 72 hours, showed pH-dependent release. At pH 8.5 (similar to that of infected wounds), an effective AgNP concentration (~35 µg/mL) was reached after ~3 hours, increasing to ~100 µg/mL after 72 hours. These results suggest that fungal β-glucans are excellent stabilizing agents for AgNPs and that the combination of β-glucans’ immunostimulatory and AgNPs’ antibacterial properties may have a synergistic effect in the treatment of infected wounds. Hydrogel containing AgNPs both prolonged, and pH-dependent release of the particles.
This work was supported by the Ministry of Science, Technological Development and Innovation of the Republic of Serbia (Contract No. 451-03-136/2025-03/ 200135, 451-03-137/2025-03/ 200135, 451-03-136/2025-03/ 200287 and 451-03-136/2025-03/200019)
The statements, opinions and data contained in the journal are solely those of the individual authors and contributors and not of the publisher and the editor(s). We stay neutral with regard to jurisdictional claims in published maps and institutional affiliations.
