Page 45 - JCTR-11-1

Journal of Clinical and
Translational Research Hesperidin enhances repair of γ-irradiated wounds

3. Results 3.2. Experiment 2: MHT

3.1. Experiment 1: Wound contraction The complete closure of wounds was observed by day
18.0 ± 0.65 days in the CMC + sham-irradiation group. In
The regeneration and repair of excision wounds can be contrast, treatment with hesperidin resulted in a significant,
effectively assessed through the periodic calculation of dose-dependent reduction in the MHT in the hesperidin
wound area as a measure of wound contraction. The + sham-irradiation group, with the maximum reduction
area of each wound at specific time points is expressed recorded for the group receiving 20 fractions of hesperidin,
as the percentage of its original size on day 1. The mean where the MHT was 15 days. Exposure to different doses of
corresponding wound area for each group was plotted as
a function of days post-wounding. The excision wound fractionated γ-radiation resulted in a dose-dependent delay
in wound closure, extending the MHT. An MHT of 21.0 ±
area progressively decreased over time, demonstrating 0.29, 24.0 ± 0.49, and 30 ± 0.33 days was observed in the
a consistent contraction of the wound in both the CMC animals exposed to 10, 20, and 40 Gy, respectively, in the
+ sham-irradiation and hesperidin + sham-irradiation CMC + irradiation group (Figure 2). Hesperidin treatment
groups (0 Gy). Maximum wound contraction was
observed on day 6 post-irradiation in both the CMC at a dose of 100 mg/kg before exposure to various doses of
and hesperidin + sham-irradiation groups (Figure 1A). fractionated γ-radiation mitigated the radiation-induced
Hesperidin treatment resulted in a significant and dose- delay in excision wound healing, resulting in MHTs of
19 ± 0.46, 22.0 ± 0.26, and 29.0 ± 0.37 days for exposures to
dependent enhancement in wound contraction at all 10, 20 and 40 Gy, respectively, in the hesperidin +irradiation
post-irradiation times compared to the CMC + sham-
irradiation group (Figure 1A). Furthermore, while the group (Figure 2). This treatment advanced MHT by
CMC + sham-irradiation group exhibited scab formation, approximately 2 days for both 10 Gy and 20 Gy, while the
this was relatively less pronounced in the hesperidin + reduction for 40 Gy was <2 days. The reductions in wound
sham-irradiation group. healing time were statistically significant (P < 0.05) for 10
and 20 Gy in the hesperidin + irradiation group.
Exposure to various doses of fractionated γ-radiation
resulted in a dose-dependent delay in wound contraction 3.3. Experiment 3: Biochemical estimations
(Figure 1A). Specifically, exposure to 10 Gy delayed The estimation of hydroxyproline levels in granulation
wound contraction at days 6 (P < 0.02) and 9 (P < 0.03) tissues serves as an accurate indicator of collagen
post-irradiation (Figure 1B), accompanied by an earlier biosynthesis during wound repair and regeneration.
scab formation compared to the CMC + sham-irradiation Hydroxyproline content increased with time, peaking on
group. The difference in wound contraction between day 8 post-irradiation in both the CMC and hesperidin
CMC + irradiation (20 Gy) and CMC + sham-irradiation + sham-irradiation groups. Following this peak, the
group was significant at days 3 (P < 0.01), 6 (P < 0.005), synthesis of new collagen declined by 12 days post-
9 (P < 0.002), 12 (P < 0.005), and 15 (P < 0.01) post- irradiation; however, hydroxyproline levels on day 12
irradiation (Figure 1C). Exposure to 40 Gy significantly remained higher than those measured on day 4 post-
delayed wound contraction at all the post-irradiation irradiation in both groups (Figure 3). The pattern of
times (Figure 1D), with scab formation being notably thick collagen synthesis was almost similar in the CMC +
compared to the CMC + sham-irradiation group. irradiation group, although exposure to 10, 20, or 40 Gy

Hesperidin treatment before exposure to 10 Gy resulted in a significant, dose-dependent decline in
irradiation significantly reduced the radiation-induced collagen synthesis at all post-irradiation times compared
delay in wound contraction on days 3 (P < 0.03), 6 to the CMC + sham-irradiation and hesperidin + sham-
(P < 0.03), 9 (P < 0.04), and 12 (P < 0.05) post-irradiation irradiation groups (Figure 3). A significant reduction in
(Figure 1B), with thinner scab formation. Furthermore, collagen deposition was observed on days 4 (P < 0.002),
hesperidin treatment before exposure to 20 Gy irradiation 8 (P < 0.001), and 12 (P < 0.002) post-irradiation for
resulted in a significant increase in wound contraction on the 40 Gy of CMC + irradiation group compared to the
days 3 (P < 0.05), 6 (P < 0.05), and 9 (P < 0.05) compared CMC + sham-irradiation group. Despite the overall
to the concurrent CMC + irradiation group (Figure 1C). reduction, maximum collagen synthesis was observed on
However, a significant increase in wound contraction after day 8 post-irradiation in the CMC + irradiation group.
hesperidin treatment before exposure to 40 Gy radiation In the hesperidin + irradiation group, the treatment with
was detected only at days 6 (P < 0.05) and 9 (P < 0.05) 100 mg/kg hesperidin before exposure to various doses of
post-irradiation in the hesperidin + irradiation group fractionated γ-radiation resulted in a significant elevation
(Figure 1D). in collagen synthesis compared to the concurrent CMC

Volume 11 Issue 1 (2025) 39 doi: 10.36922/jctr.24.00049