Che Soh N' et al. (2020), Acute Wound Healing Potential of Marine Worm, D...

ECB-ART-49354

Evid Based Complement Alternat Med 2020 Nov 25;2020:6688084. doi: 10.1155/2020/6688084.

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Acute Wound Healing Potential of Marine Worm, Diopatra claparedii Grube, 1878 Aqueous Extract on Sprague Dawley Rats.

Che Soh N' , Rapi HS , Mohd Azam NS , Santhanam RK , Assaw S , Haron MN , Ali AM , Maulidiani M , Idris I , Ismail WIW .

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Diopatra claparedii which is colloquially known as Ruat Sarung can be found along the west coast of Peninsular Malaysia. The species has a unique ability to regenerate anterior and posterior segments upon self-amputation or injury, thus having potential as a wound healing promoter. In this study, the wound healing potential of D. claparedii aqueous extract on acute wound model in rats was revealed for the first time. Various concentrations (0.1%, 0.5%, and 1.0% w/w) of D. claparedii ointment were formulated and tested on Sprague Dawley rats through topical application on full-thickness skin wounds for 14 days. The wound healing effects were investigated via behaviour observation, wound contraction, and histopathological analysis. Quality assessment was performed via skin irritation test, microbial contamination test (MCT), and heavy metal detection. The study also included test for antibacterial activities and detection of bioactive compounds in D. claparedii. One percent of D. claparedii ointment showed rapid wound healing potential with good soothing effects and more collagen deposition in comparison to the commercial wound healing ointments such as acriflavine (0.1% w/v) and traditional ointment gamat (sea cucumber extract) (15.0% w/v). No local skin irritation, microbial contamination, and insignificant concentration of heavy metals were observed, which indicate its safe application. Moreover, the aqueous extract of D. claparedii exhibited antibacterial activities against Escherichia coli and Pseudomonas aeruginosa with minimum inhibitory concentration (MIC) value at 0.4 g/ml. 1H NMR analysis of the aqueous extract of D. claparedii revealed some metabolites that might be responsible for its wound healing properties such as amino acids, halogenated aromatics, organic acids, vitamins, and others. Altogether, these results suggested that the aqueous extract of D. claparedii could be utilised as an alternative natural wound healing promoter.

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	Figure 1. Gross observations of wound healing progression of 8 mm in diameter of excisional wound on dorsum of Sprague Dawley rat. Seven treatment groups were tested which were 0.1%, 0.5%, and 1.0% of polychaete ointment (PO), gamat (15%), acriflavine (0.1%), cetomacrogol emulsifying ointment (negative control), and untreated group. The observation showed the wound was healed gradually within 14 days of experimental period (n = 5).
	Figure 2. Wound healing percentage of different treatment groups measured on days 3, 7, 11, and 14 after wounding. The wound contraction was measured as percentage reduction from original wound size. Seven treatment groups were tested which were 0.1%, 0.5%, and 1.0% of polychaete ointment (PO), gamat (15%), acriflavine (0.1%), cetomacrogol emulsifying ointment (negative control), and untreated group. The result showed that wound treated with 1.0% PO demonstrated rapid wound closure as compared to other treatment groups. Data are mean ± SEM (n = 5). The significance difference was analysed using one-way ANOVA. ∗p < 0.05 was compared between 1.0% PO, negative control, and untreated group.
	Figure 3. Hematoxylin & Eosin (H & E) staining of granulation healing tissue of Sprague Dawley rats treated with different treatment groups on day 14 after wounding. Seven groups were tested which were (a) untreated group, polychaete ointment (PO) with concentration of (b) 0.1%, (c) 0.5%, and (d) 1.0%, (e) cetomacrogol emulsifying ointment (negative control), (f) gamat 15%, and (g) acriflavine 0.1%. Smaller granulation tissue with denser collagen fibres and more new capillaries formation was seen in 1.0% PO than other treatments. E = epidermis, S = scar/granulation tissue, N = normal dermis, H = hair follicle, B = blood vessel. 40X magnification.
	Figure 4. Masson's trichrome (MT) staining of granulation healing tissue of Sprague Dawley rats treated with different treatment groups at day 14 after wounding. Seven groups were tested which were (a) untreated group, polychaete ointment (PO) with concentration of (b) 0.1%, (c) 0.5%, and (d) 1.0%, (e) cetomacrogol emulsifying ointment (negative control), (f) gamat 15%, and (g) acriflavine 0.1%. New collagen formation was seen as FC and matured collagen as CC. More new collagen depositions in meshwork formation appeared in 1.0% PO. S = scar/granulation tissue, E = epidermis, CC = coarse collagen, FC = fine collagen, H = hair follicle. 40X magnification.
	Figure 5. 1H NMR spectrum of the aqueous extract of D. claparedii in DMSO-d6 containing 0.03% tetramethylsilane TMS.

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