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Full Research Article
Nile Tilapia Skin as Dermal Wound Healing Promoter in Cats
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During recent years there is increasing trend to use tilapia fish for different medical purposes (Ibrahim et al. 2020). Tilapia fish skin shows a number of unique characteristic and the most important of them is that it has thick slime coated skin that serves as a barrier for many bacterial infections.The simplicity in breeding tilapia is that it grows well in high density and this makes it suitable for farming (DeLong et al., 2009). Tilapia fish has great economic impact as it is in greater number in inland waters and can be cultivated easily (Yue et al., 2016). The scientific name of Nile tilapia is Oreochromis niloticus and is commonly named “tilapia”. Tilapia (Oreochromis niloticus) is a fresh water most cultured fish in the world.
Skin of tilapia fish has unique property which separates it from other is that it contains gelatin that can be used in emulsions and in few recent years this property has made tilapia in medicine field a center of attraction for everyone. (Zhang et al. 2020). Hydrolysates of gelatin obtained from tilapia fish skin proved to have dynamic antioxidant and anti-hypertensive effects (Choonpicharn et al. 2015). Tilapia fish skin possesses antiviral and antibacterial properties. As it provides a barrier for bacteria for 24-48 hours and this antibacterial barrier activity of fish skin is tested by a 2-chamber model. Tilapia fish skin is preferred to use for wound healing because it prevents bacteria to invade the wound. Tilapia skin has greater ability to support the growth of cells on wound area in 3 dimensions (Magnusson et al. 2017). Another use of Tilapia fish skin is as a biomaterial used for dressing in case of burn wounds because it is rich in collagen that promotes wound healing. High level of collagen in tilapia fish skin makes it more biocompatible to be used as graft. Collagen present in tilapia fish skin shows quick absorption on wound site as compared to bovine and procain collagen. It has gained hype in cosmetic industry too because COL-1 from tilapia fish skin is safest to use and has no adverse effects (Verde et al. 2021).
Widespread application of tilapia fish skin grafting has been reported in human medical practice following its approval by FDA. Tilapia fish skin has properties that enhance the process of wound healing. Due to its cost effectiveness these grafts are potential alternates of allograft and xenograft. Besides wound healing these grafts have shown encouraging results in treating diabetes foot ulcers, venous leg ulcers and other acute and chronic wounds. By using tilapia fish skin there are very less chances of transmission of any viral disease to other species and that is why preparation of fish skin as a graft requires less effort (Fiakos et al., 2020).
According to a report tilapia fish skin graft has resulted healing in 6.5 million patients in US by promoting wound healing to live a healthier life. Tilapia fish skin is also known for possessingvarious bioactive properties. It acts as bacterial barrier and promotes 3-dimensional (3-D) growth of cells on wound site. A random clinical trial study was conducted on 162 full, thickness wounds by using tilapia fish skin as it was noted that wound with tilapia fish skin healed within 14 days of application. After 14 days 13% of the fish skin treated group had healed as compared to the 6% procain SIS matrix treated group. After 21 days 72.5% of fish skin treated group had healed and 65%of the other group. Collectively the study showed that fish skin helps wound to heal faster. This study also showed that fish skin do not actives any autoimmune activity of body again the fish skin graft (Magnusson et al. 2016).
In human reconstructive surgical practice tilapia skin is now considered a great alternative for skin grafting. The structure of tilapia fish skin is highly porous, with high level of hydrophilicity, property to absorb water and greater permeability. All these properties make tilapia skin a good alternative of synthetic skin grafts as it promotes wound healing by enhancing angiogenesis and collagen deposition on wound site (Chen et al., 2021).
Currently, fish collagen is being used widely in different biomedical applications linked with wound healing, tissue engineering, regeneration, drug delivery, cell culture and other therapeutic procedures (Rodríguezet_al2020). Nile tilapia fish skin is also proved to be effective when used as xenograft for the treatment of burn wounds (Dias et al., 2020).
Tilapia fish skin accelerates the wound healing by decreasing the effects of pro-inflammatory (TNF-α, IL-6 and IL-8) and increase the number of receptors on target cells for β-defense 14, pattern recognition receptor (NOD2), anti-inflammatory (IL-10), vascular endothelial growth factor (VEGF) and fibroblast growth factor (β-FGF). Excellent healing is obtained due to high content of collagen in tilapia fish skin and by its activity to increase the growth of bacteria that promotes wound healing and decrease the bacteria that disturbs the process of wound healing (Mei et al., 2020b).
So far no study data is available on wound healing properties of Nile tilapia skin in veterinary practice. The objective of this study was to evaluate the efficacy of Nile tilapia skin as wound healing promoter biological dressing in cats.
MATERIALS AND METHODS
The present study was performed at Department of Small Animal Clinical Sciences, University of Veterinary and Animal Sciences, Lahore, Pakistan in 2022. The study was conducted on randomly selected 10 vaccinated clinical cases of cats having minor or major losses of skin. Study included cats of different breed, age and sex.
Fully vaccinated, fresh wounds with major or minor skin losses are included.
Old necrosed infectious wounds, more than 7 days old dog/cat bite wound, wounds because of stray cat bites were not included, poor health condition with compromised immune system, underlying severe disease or zoonotic infection, pregnancy are not included.
Selected cats were divided randomly into 2 groups i.e. A and B each with 5 cats and subjected to the treatment plan as follows (Table 1).
Fresh fish was collected from Tilapia hatchery and transported to UVAS Department of Small Animal Clinical Sciences in thermostat container having ice in it. Skin of fish was separated from muscles with the help of sharp scalpel blade. Harvested skin was then washed with sterile normal saline (0.9% NaCl) to remove debris and impurities. Collected skin was placed in a sterile container filled with normal saline (Ibrahim et al., 2020a). Collected fish skin strips were dipped in 10% povidone iodine solution for 10-15 minutes before application.
• Wound was washed with normal saline followed by disinfection with 10% povidone iodine.
• Skin edges of wound were trimmed in cosmetic appearance for regular pattern.
• Gas anesthesia was used by using isoflurane.
• Wound size was measured using vernier caliper.
Fish skin was trimmed and shaped according to the shape and size of recipient site. The dressing was applied on wound as per following technique.
Dressing was secured on wound site by applying simple interrupted sutures with non-absorbable suture material (silk 2-0) (Fig 1). The dressing was covered with gauze impregnated with paraffin to avoid adherence of tilapia skin with bandage. Gauze and wound were secured with simple adhesive tape.
Wound area was washed with 0.9% Normal Saline (NaCl) solution and treated as an open wound.
Parameters of study
Parameters of study includedwound size, physical characteristics of wound and Quality of healing (presence or absence of scar formation, speed of healing, sensitivity response to tilapia skin dressing, histo-pathological evidences) showing wound healing process and skin regeneration from day 0 to 14 and sterility of wound).
Biopsy sample was collected using 3mm punch biopsy on days 0, 3, 7 and 14 to evaluate the histo-pathological findings at various intervals during wound healing (Parente et al., 2012) Collected samples were preserved in formalin while tissue processing was performed in the Department of Pathology, University of Veterinary and Animal Sciences, Lahore, Pakistan.
The data thus obtained was statistically analyzed by using independent T test on SPSS version 20.
RESULTS AND DISCUSSION
Following were the findings of present study in group A and B on the basis of pre-selected parameters.
Wound size was measured in each cat of both groups on days 0, 3, 7 and 14. In group A, on days 0, 3, 7 and 14 mean wound size was 73.92±30.20, 41.24±15.26, 25.36±13.61 and 12.2±9.23, respectively. Statistical analysis revealed significant difference (P<0.05) among means of wound size changes during healing from days 0-14. In group B, on days 0, 3, 7 and 14 mean wound size was 75.40±29.91, 69.40±27.55, 61.52±26.61 and 54.44±23.04, respectively. Statistical analysis showed a significant difference (P<0.05) among means of wound healing area in group B from days 0-14. According to Guo and DiPietro (2010) skin wound healing is a naturally complicated biological mechanism that involves highly programed stages i.e. inflammation, proliferation and remodeling.
Physical characteristics of wound
Both groups A and B included cats with different characteristics of skin wound in terms of shape, size and healing status (Table 2).
In all cases wounds were not more than 24 hours old. In group A out of total 5 cats 3 cats were wounded due to cat fight cases where as other 2 cats were accidental injuries. In group B, out of total 5 cases 3 were the cat fight cases where as two cases involved accidental injuries (Table 3).
Quality of wound healing
Quality of wound healing was evaluated on the basis of following factors.
a. Scar formation
In both groups A and B, dressing was changed after every 48 hours to avoid contamination of site. In group A, out of total 5 cats no cat showed scar formation. In group B, out of total 5 cats 3 cats showed scar formation and in rest of the 2 cats no scar formation took place mainly because Tilapia fish skin accelerates wound healing by decreasing the effects of pro-inflammatory mediators (TNF-α, InterLeukin-6 and InterLeukin-8) and it increases the number of receptors on target cells for β-defense 14, pattern recognition receptor (NOD2), anti-inflammatory (IL-10), vascular endothelial growth factor and fibroblast growth factor (β-FGF) as observed by Mei et al., (2020a).
b. Sensitivity raction
In both groups A and B, during the entire course of study, no cat showed any sign of sensitivity reaction to tilapia skin dressing. According to Magnusson et al. (2016) tilapia fish skin does not active any autoimmune activity of body again the fish skin graft that is why no sensitivity reaction is noted. These findings are also in line with the findings of Ibrahim et al., (2020). They reported thatTilapia fish skin is a rich source of gelatin as well as collagen.It possesses biocompatible type 1 collagen which has potential for using as clinical regenerative procedure to treat massive dermatological losses smoothly.
c. Speed of healing
According to statistical analysis in group A, the data on the relation between mean wound size with respect to days of complete healing showed a significant difference (P<0.05) that indicates speedy wound healing process. Graphical presentation with respect to statistical data also demonstrated that during in most of the wounds maximum healing was achieved by day 14. In group B, according to statistical analysis, data on relation between mean wound size of this group with respect to days of complete healing showed a significant difference (P<0.05) that indicate wound healing process. Graphical presentation with respect to statistical data showed that during trial in most of wounds delayed healing was achieved as compared to group A. Same observations are documented by Mei et al., (2020a). They documented that Tilapia fish skin accelerates wound healing by decreasing the effects of pro-inflammatory mediators (TNF-α, InterLeukin-6 and Inter Leukin-8) and it increases the number of receptors on target cells for β-defense 14, pattern recognition receptor (NOD2), anti-inflammatory (IL-10), vascular endothelial growth factor and fibroblast growth factor (β-FGF). It increases the bacterial growth responsible to promote wound healing and decreases that bacterial growth which disturbs wound healing.
d. Histo-pathological findings
Histo-pathological evaluationin group A, revealed a fast healing process followed by the complete wound healing by day 14 in all cats of this group (Fig 2).
Fig 2: Histo-pathological findings in group A: An absence of epidermis observed on day 0. On day 3, no changes observed in the dermal area of the section. No epidermis seen except in the middle of the section. In the peripheral area the epidermal layer was thin and regenerating. Small amount of dead tissue mass was observed over the affected area. On day 7, active cells of stratum basale were seen dividing. Layers of stratum spinosum had formed. Epithelialization was in progress. On day 14, Epithelialization was at its peak. Multiple layers of squamous epithelial cells had deposited. Mature keratin was visible in the stratum corneum. Kerato-hyalin granules in the stratum granulosum were large and numerous.
Based on the histo-pathological evaluation in group B, the healing process was slow and wound healing was incomplete in all cats on day 14 (Fig 3).
Fig 3: Histo-pathological findings in group B: A complete absence of epidermis on day 0. On day 3, dead tissue mass observed along with the dead and dying neutrophils present in the section. On day 7, epidermal regeneration was noted. On day 14, the process of epithelialization was still incomplete and minimal keratin was seen on the upper layer.
Wound size comparison between Group A and B
Statistical analysis based comparison of wound size on days 0-14 in group A and B revealed a significant difference (P<0.05) both groups with respect to wound size at different intervals (Fig 4).
Conflict of interest
- Chalkowski, K., Wilson, A.E., Lepczyk, C.A., Zohdy, S. (2019). Who let the cats out? A global meta-analysis on risk of parasitic infection in indoor versus outdoor domestic cats (Felis catus). Biology Letters. 15(4): 20180840.
- Chen, H., Yin, B., Hu, B., Zhang, B., Liu, J., Jing, Y., Fan, Z., Tian, Y., Wei, X., Zhang, W. (2021). Acellular fish skin enhances wound healing by promoting angiogenesis and collagen deposition. Biomedical Materials. 16(4): 045011.
- Choonpicharn, S., Jaturasitha, S., Rakariyatham, N., Suree, N., Niamsup, H. (2015). Antioxidant and antihypertensive activity of gelatin hydrolysate from Nile tilapia skin. Journal of Food Science and Technology. 52(5): 3134-3139.
- DeLong, D.P., Losordo, T., Rakocy, J. (2009). Tank culture of tilapia. SRAC Publication no. 282.
- Dias, M.T.P.M., Bilhar, A.P.M., Rios, L.C., Costa, B.A,, Júnior, E.M.L., Alves, A.P.N.N., Bruno, Z.V., de Moraes, Filho M.O., Bezerra. L.R.P.S. (2020). Neovaginoplasty using Nile Tilapia fish skin as a new biologic graft in patients with Mayer-Rokitansky-Küster-Hauser syndrome. Journal of Minimally Invasive Gynecology. 27(4): 966-972.
- Fiakos, G., Kuang, Z., Lo, E. (2020). Improved skin regeneration with acellular fish skin grafts. Engineered Regeneration. 1: 95-101.
- Guo, Sa., DiPietro, L.A. (2010). Factors affecting wound healing. Journal of Dental Research. 89(3): 219-229.
- Ibrahim, A., Hassan, D., Kelany, N., Kotb, S., Soliman, M. (2020). Validation of three different sterilization methods of Tilapia skin dressing: Impact on microbiological enumeration and collagen content. Frontiers in Veterinary Science. 7: 597751.
- Magnusson, S., Baldursson, B.T., Kjartansson, H., Rolfsson, O., Sigurjonsson, G.F. (2017). Regenerative and antibacterial properties of acellular fish skin grafts and human amnion/ chorion membrane: Implications for tissue preservation in combat casualty care. Military Medicine. 182(1): 383- 388.
- Magnusson, S., Winters, C., Baldursson, B.T., Kjartansson, H., Rolfsson, O., Sigurjonsson, F. (2016). Acceleration of wound healing through utilization of fish skin containing omega-3 fatty acids. Today’s Wound Clinic. Pg. 26-29.
- Mei, F., Duan, Z., Chen, M., Lu, J., Zhao, M., Li, L., Shen, X., Xia, G., Chen, S. (2020a). Effect of a high-collagen peptide diet on the gut microbiota and short-chain fatty acid metabolism. Journal of Functional Foods. 75: 104278.
- Mei, F., Liu, J., Wu, J., Duan, Z., Chen, M., Meng, K., Chen, S., Shen, X., Xia, G., Zhao, M.J.J.oa., chemistry f. (2020b). Collagen peptides isolated from salmo salar and tilapia nilotica skin accelerate wound healing by altering cutaneous microbiome colonization via upregulated NOD2 and BD14. 68(6): 1621-1633.
- Parente, L.M.L., Lino Júnior, Rd.S., Tresvenzol, L.M.F., Vinaud, M.C., de Paula, J.R., Paulo, N.M. (2012). Wound healing and anti-inflammatory effect in animal models of Calendula officinalis L. growing in Brazil. Evidence-based Complementary and Alternative Medicine. DOI: 10.1155/2012/375671.
- Rodríguez, Á.H., Júnior, E.M.L., de Moraes, Filho, M.O., Costa, B.A., Bruno, Z.V., Monteiro Filho M.P., de Moraes, M.E.A., Rodrigues, F.A.R., Paier, C.R.K., Bezerra, L.R.P.S. (2020). Male-to-female gender-affirming surgery using Nile tilapia fish skin as a biocompatible graft. Journal of Minimally Invasive Gynecology. 27(7): 1474-1475.
- Verde, M.E.Q.L., Ferreira-Júnior, A.E.C., de Barros-Silva, P.G., de Castro Miguel. E,, Mathor. M.B., Lima-Júnior, E.M., de Moraes-Filho, M.O., Alves, A.P.N.N. (2021). Nile tilapia skin (Oreochromis niloticus) for burn treatment: Ultrastructural analysis and quantitative assessment of collagen. Acta Histochemica. 123(6): 151762.
- Yue, G., Lin, H., Li, J. (2016). Tilapia is the fish for next-generation aquaculture. International Journal of Marine Science and Ocean Technology. 3(1): 11-13.
- Zhang, T., Sun, R., Ding, M., Tao, L., Liu, L., Tao, N., Wang, X., Zhong, J. (2020). Effect of extraction methods on the structural characteristics, functional properties and emulsion stabilization ability of Tilapia skin gelatins. Food Chemistry. 328: 127114.
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