Fig 1.
This figure shows the hydrogel film placed within a petri dish, demonstrating its uniform structure and translucent appearance post-synthesis.
Fig 2.
Characterization of SA-CMC-HA hydrogels.
(A) Water Absorption Rate in SA-CMC-HA Hydrogel In 7 days. (B) Swelling Ratio in SA-CMC-HA Hydrogel In 7 days.
Fig 3.
Degradation profile of SA-CMC-HA hydrogel over seven days.
The figure presents the degradation behavior of the SA-CMC-HA hydrogel over a period of seven days, measured as the percentage of hydrogel mass remaining (%). The degradation shows a gradual decline in hydrogel mass over time, which decreases steadily until Day 7. This profile indicates the hydrogel’s stability and controlled biodegradability, suitable for applications requiring a slow degradation rate.
Fig 4.
Surface morphology at a scale of 50 μm.
(A): Control hydrogel with an almost uniform and smooth surface. (B): Hydrogel infused with bacteriophage SAM-E.f 12 exhibiting a protruding surface texture.
Fig 5.
FTIR spectrum of (A). SA, (B). CMC, (C). HA, and (D). SA-CMC-HA Hydrogel. The peaks in the range of 3200–3500 cm-1 are related to (O-H), those in the 1620–1420 cm-1 range to (C = O and C-N) bonds, and those at 1020 cm-1 to (C-O) bonds. The presence of distinctive peaks at 2330 cm⁻¹ in the spectra of the phage-loaded hydrogel indicates the possible retention of phages within the hydrogel.
Fig 6.
(A) This graph shows that the direct phage and phage-loaded hydrogel effectively suppressed bacterial growth, as evidenced by the steady decrease in optical density (OD) of the bacterial culture over time compared to the control. (B) BHI broth contains only Enterococcus faecalis (Control), phage-containing hydrogel, and direct phage. The effectiveness of the hydrogel-containing phage and direct phage in a liquid culture medium is illustrated, with the inhibitory effect contrasting sharply with the control group (no hydrogel), which showed a significant increase in OD.
Fig 7.
(A) A disk infused with hydrogel-containing phage produced a clear halo around the surrounding bacteria. (B) Blank hydrogel discs were used as negative controls, and no halo formation was observed. (C) Direct phage produced a clear halo around the surrounding bacteria.
Fig 8.
The biostability of phage SAM-E.f 12 in SA-CMC-HA hydrogel was evaluated after storage for 0, 1, 2, 3, and 4 weeks at 25°C.
The phage count was determined using double-layer agar for conducting plaque assay (n = 3), and the error bars represent the standard deviations.
Fig 9.
(A) Comparative Analysis of Treated Groups. The chart illustrates the bacterial load in different groups of mice on days 1, 3, 7, 10, and 14. Wound samples were collected at specific intervals. (B) The group treated with pure hydrogel (without phage), (C) The group treated with hydrogel containing phage, (D) The group treated with direct phage.
Fig 10.
(A) Effect of Phage-Containing Hydrogel on Wounds of Male BALB/c Mice Infected with E. faecalis. (II) Negative wound control group (without bacterial infection, no treatment). (III) The positive control (wound, with bacterial infection, and no treatment). (IV) The group was treated with a hydrogel containing bacteriophage (wound infected with E. faecalis). (V) The group was treated with a hydrogel without phage (wound, infected with E. faecalis). (VI) The group treated with direct phage (wound and infected with E. faecalis). (B) Wound healing rates based on wound sizes measured over 14 days in different treatment groups of mice. The graph represents the following groups: II (Wounded negative control): Mice without treatment (blue line). III (Positive control): Mice treated with standard wound healing agents (orange line). IV (Phage-containing hydrogel): Mice treated with hydrogel infused with bacteriophages for enhanced wound healing (gray line). V (Hydrogel without phage): Mice treated with hydrogel alone, without bacteriophages (yellow line). VI (Direct phage): Mice treated with bacteriophages applied directly to the wound (light blue line). All groups’ wound sizes decrease over time, with the fastest reduction observed in the Phage-containing hydrogel group (IV) and the slowest in the Wounded negative control group (II). Time points include measurements on days 1, 3, 7, 10, and 14.
Fig 11.
Histopathological evaluation of wound healing outcomes.
Sections of the skin in different groups with 10x magnifications. The areas marked with blue rectangles indicate the locations of the wounds. (A) I) control group (healthy mice). (B) II) Negative control group (wounded mice, without bacterial infection, without treatment). (C) III) Positive control group (wounded mice, wounds infected by E. faecalis bacteria, without treatment). (D) IV) The group was treated with a hydrogel containing SAM-E.f 12 phage (wounded mice, the wounds were infected with E. faecalis). (E) V) The group was treated with a hydrogel without phage (wounded mice, the wounds were infected with E. faecalis). (F) VI) The group was treated with direct phage (wounded mice, the wounds were infected with E. faecalis).
Table 1.
Histopathological assessment of tissue parameters collected from wounds on day 14.