Page 42 - JCTR-11-1

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

2. Materials and methods 100 mg/kg body weight hesperidin once daily, depending
on the total fractionated irradiation dose. A gap of 2 days
2.1. Animals was consistently maintained between the delivery of the
Animal care and handling were conducted following the 6 , 11 , or 16 fractions of the radiation dose.
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guidelines of the World Health Organization, the Indian
National Science Academy, and the “Guide for the Care 2.4. Experimental protocol
and Use of Laboratory Animals” (NIH publication #86- The effect of hesperidin treatment after delivery of
23, revised in 1985). Swiss albino mice, aged 8 – 10 weeks different doses of fractionated radiation was investigated
of either sex and weighing 30 – 36 g, were sourced from by performing the following experiments:
an inbred colony maintained under controlled conditions
of temperature (23 ± 2°C), humidity (50 ± 5%), and 2.4.1. Experiment 1: Wound contraction
light (12 h of light and dark cycle). The animals had free Wound contraction is an important aspect of wound repair
access to sterile food and water. The diet consisted of 50% and regeneration, which was studied by dividing an equal
cracked wheat, 40% Bengal gram, 4% milk powder, 4% number of males and females into the following groups:
yeast powder, 0.75% sesame oil, 0.25% cod liver oil, and (i) CMC + sham-irradiation: Animals in this group
1% salt. Four animals were housed in a polypropylene received 0.01 mL/g body weight of 1% CMC once
cage containing sterile paddy husk (procured locally) daily, consecutively for 5, 10, or 20 days before sham-
as bedding throughout the experiment. The study was irradiation.
approved by the institutional animal Ethical Committee of (ii) Hesperidin + sham-irradiation: Animals in this
Manipal University, Manipal, India (Approval no.: IAEC/ group were administered 100 mg/kg body weight
KMC/03/2003-2004). of hesperidin once daily, consecutively for 5, 10, or
20 days before sham-irradiation.
2.2. Chemicals
(iii) CMC + irradiation: This group received 0.01 mL/g
Hesperidin (catalog no.: 12346-0050), hydroxyproline body weight of 1% CMC once daily, consecutively for
(catalog no.: H5534), chloramine T (catalog no.: C9887), 5, 10, or 20 days before exposure to various doses of
glucosamine (catalog no.: G44875), acetylacetone γ-radiation.
(catalog no.: A3511), deoxyribonucleic acid (catalog (iv) Hesperidin + irradiation: Animals in this group were
no.: D4522), diphenylamine (catalog no.: D2385), administered 100 mg/kg body weight of hesperidin
5,5’-dithiobis(2-nitrobenzoic acid) (DTNB, catalog once daily, consecutively for 5, 10, or 20 days
no.: D8130), ethylenediaminetetraacetic acid (EDTA, before exposure to various doses of fractionated
catalog no.: E4884), β-nicotinamideadenine dinucleotide γ-radiation.
phosphate (catalog no.: N1630), N-(1-Naphthyl)
ethylenediamine dihydrochloride (NEDD, catalog no.: 2.4.1.1. Irradiation
N5889) and sulfanilamide (catalog no.: S9251) were One hour after each administration of CMC or hesperidin,
procured from Sigma Chemical Co., United States. The each animal was placed into a specially designed well-
trichloroacetic acid (TCA, catalog no.: 15213-5000) ventilated acrylic restrainer. The lower half of the animals
and ρ-dimethylaminobenzaldehyde (Ehrlich’s reagent (below the rib cage) was exposed to a single dose of either
[catalog no.: 42363-0250]) were requisitioned from Across 0 or 2 Gy once daily, delivered at a dose rate of 1.35 Gy/min
Organics, Belgium. Imidazole buffer (catalog no.: 38546) from a Co Teletherapy source (Theratron, Atomic Energy
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was procured from S.d. fine-chemicals Ltd., India. Ethanol Agency, Canada). Treatments were given once daily for
(catalog no.: SIN 1170) was purchased from Hayman 5, 10, or 20 days, resulting in a total dose of 10 Gy (five
Ltd., England, and sterillium disinfectant solution was fractions of 2 Gy each), 20 Gy (10 fractions of 2 Gy each),
supplied by Bode Chemie, Germany. Perchloric acid, or 40 Gy (20 fractions of 2 Gy each). A gap of 2 days was
carboxymethylcellulose (CMC), diethyl ether, formalin, consistently maintained between the delivery of the 6 ,
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sodium hydroxide, sodium nitrate, sodium nitrite, 11 , or 16 fractions of the radiation dose.
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phosphoric acid, hydrochloric acid (HCl), and sodium
chloride were obtained from Ranbaxy Fine Chemical, India. 2.4.1.2. Production of full-thickness skin wound
The fur of the dorsum of each animal was removed
2.3. Preparation of drug and mode of administration with a cordless electric mouse clipper (Wahl Clipper
The required amount of hesperidin was suspended in Corporation, United States) before exposure to the last
1% CMC, and the animals were administered orally fraction of γ-radiation. A full-thickness skin wound was
with a single dose of 0.01 mL/g body weight of CMC or created on the dorsum (below the rib cage) as previously

Volume 11 Issue 1 (2025) 36 doi: 10.36922/jctr.24.00049