Honey is one of the oldest substances used in wound management. Efficacy of Gelam honey in wound healing was evaluated in this paper.
Wound healing is a complex biological cascade of cellular and biochemical events comprised of three phases: inflammation, proliferation, and maturation [
Honey has been used for its medicinal properties in many cultures since ancient times [
In Malaysia, a variety of honey is locally produced. Unfortunately, there is still little evidence to support the potential of Malaysian honey on wound healing. Thus, this present study was designed to evaluate the efficacy of a selected Malaysian honey towards excisional wound healing.
Gelam honey is a local monofloral honey produced by
This wound dressing is a clear amorphous hydrogel containing modified carboxymethyl cellulose polymer, propylene glycol, and water. It provides a moist environment which is suitable for wound healing and promotes natural debridement by rehydrating necrotic tissue. Intrasite Gel (Smith & Nephew) was obtained from the pharmacy of University of Malaya Medical Centre.
Adult male
Uninfected, 4 cm2 full thickness, excisional wounds were used in this study to represent the acute wound healing (Figure
2 cm by 2 cm excision skin wound on day 0, before application of treatments.
This experimental protocol was adapted and modified from Aljady et al. [
The wounding procedure was carried out under general anesthesia by using Ketamine 50 mg/kg and Xylazine 5 mg/kg in mix in same syringe-injected intramuscularly, producing 30 minutes of anesthesia. Dorsum area of the rats was shaved by electrical hair clipper and swabbed with 70% alcohol and 1 mL of Lignocaine HCl (2%, 100 mL/5 mL) was injected subcutaneously for local anesthesia. A square wound (2 cm by 2 cm) was marked by using wound mold on the posterior neck area between the shoulders and excised carefully by using a pair of surgical scissors aseptically. The wound mold did not contact the wound, since it was only used in marking the border of the wound before the surgery. The location of the wound area was chosen so as to prevent unwanted wound caused by stretching and biting from the rat, itself. The mold, scissors, and forceps were cleansed with 95% alcohol after each use. All wounds were of full thickness extending vertically down to the subcutaneous tissue.
Treatments started 24 hours after wound creation, the wounds were air exposed, and dressing was applied topically once a day. The rats were randomly divided into four experimental groups with 24 animals per group. Untreated group was left without any treatment to serve as the untreated control group. Saline group was treated topically with normal saline (negative control), Intrasite group was treated with Intrasite Gel (positive control), while Gelam group (experimental group) was treated with Gelam honey.
Six rats from each group were sacrificed at days 1, 5, 10, and 15 of treatments. Entire wound tissue area was removed carefully from each rat and fixed immediately for histological process. All samples were properly labeled with unique numbers before storage and the measurements were done at random to overcome experimental bias.
Photograph of each wound was taken for analytical purposes, and macroscopic evaluation (e.g., wound appearance) was recorded. All wounds were assessed clinically according to the scoring system modified from the clinical judgment by Bates-Jensen [
On days 1, 5, 10, and 15 of treatments, relevant groups had their wounds measured before tissues were excised. Animals were anesthetized during the procedures. Margin of the wounds was traced on a transparency paper by a fine tip permanent marker for the evaluation of the rate of wound contraction. Contraction of both longitudinal (length) and transverse (width) measurements of wounds was recorded. Wound contraction was calculated as described by Aljady et al. [
Excised tissue was fixed in 10% formalin before histologically processed. Sections were made at the thickness of 5
All values were reported as Mean ± Standard Error Mean (S.E.M.). The statistical differences among groups were assessed using one-way ANOVA (analysis of variance). A value of
The effects of various treatments on duration of wound healing process were shown in Table
Time required for wound healing in rats.
Type of dressing and grouping | Healing time (days) |
---|---|
No treatment (untreated group) | |
Normal saline (saline group) | |
Intrasite Gel (Intrasite group) | |
Gelam honey (Gelam group) |
All values were expressed as mean and standard error mean (S.E.M.); a value of
On day 1 of treatments (Figure
Excisional wound appearance after treatments: Results from (a) day 1 of treatments, (b) day 5 of treatments, (c) day 10 of treatments, and (d) day 15 of treatments; (i) untreated group, (ii) saline group, (iii) Intrasite Gel group, and (iv) Gelam honey group.
The histological sections demonstrated the process of healing reflected by the macroscopical evaluation. Figure
Photomicrographs of wound tissues at day 10 of treatments stained with H & E, 20x magnification: (a) Untreated Group, (b) Saline Group, (c) Intrasite Group, (d) Gelam Group. S—Scab; E—Epidermis; D—Dermis; GT-Granulation tissue. The arrow showed epithelialization.
The results showed a trend of progressive contraction from day 5 onwards for both longitudinal and transverse wound contraction. Percentage of wound area contraction of each treatment was shown in Figure
Contraction of wound area as percentage of original wound.
As expected, there was no significant difference (
Simple and reproducible wound model is essential for wound research. Incisional and excisional wounds are the two main wound models in wound research which allowed the determination of the wound healing phases. Full thickness excisional wounds were used in this study to macroscopically and histologically evaluate the efficacy of topical application of Gelam honey (a Malaysian honey) in facilitating wound healing. The excisional wound was found to be more suitable for histological evaluation due to the broader morphological changes occurring during the process [
The results of this study showed that topical application of Gelam honey significantly accelerated the rate of wound healing compared to the saline treated group. This is similar to findings on the efficacy of topical application of honey in wound management as reported by many researchers [
This study demonstrated that topical application of Gelam honey reduced the hard, intact dark brown scab in wound healing if compared to other treatment groups (Figure
Wound contraction is an essential process in healing that lead to wound closure. Thus, visible appearances and measurements of wound contraction become reliable parameters in macroscopic evaluation for wound healing [
Histological evaluation showed significant epithelial regeneration appearance in Gelam-honey-treated wound. The epithelialization process was almost complete in Gelam honey treatment compared to other treatment groups (Figure
This study demonstrated the efficacy of Gelam honey in wound healing. Topical application of Gelam honey accelerated the rate of wound healing by increasing the wound contraction. The acceleration of wound healing was demonstrated by the healing rate and wound contraction through macroscopic and histological evaluation. Possibly, the relevant actions regarding the healing (wound contraction percentage) seemed to be occurring between days 5 and 10. Hence, Gelam honey is potentially useful in dressing wounds but further studies are needed to detail out the specific mechanisms of the honey.
All authors have nothing to disclose and have no commercial or financial interest in the products described in this paper.
The authors are grateful to the University of Malaya for research grants PS280/2009B and RG081/09AFR. They would also like to thank the staff of Animal House, faculty of Medicine, University of Malaya, for the care and supply of rats; Mr. Pouya Hassandarvish for his help in animal work; Madam Puah Lin Eng for histotechnical consultancies; and members of Honey Research Group of University of Malaya and members of Neuroscience Research Group of University of Malaya for technical support. This work was presented in The 3rd International Conference on Natural Products for Health and Beauty: “Through Sustainable Health” on 16–18 March 2011 at The Emerald Hotel, Bangkok, Thailand.