Global Tilapia Fish Skin Treatment of Third-Degree Skin Burns: News Brief By Spherical Insights and Consulting
Tilapia skin is a visually appealing and practical therapy option for individuals with second and third-degree burns. According to numerous studies, the antibacterial and anti-inflammatory qualities of tilapia skin aid in wound healing and lessen burn patients' discomfort and inflammation. Furthermore, tilapia skin is easily accessible, reasonably priced, and unlikely to spread illness. Few researches have been conducted to elucidate the therapeutic mechanisms underlying the application of tilapia fish skin, despite the therapy's apparent promises. In this study, 16 mice with third-degree burns were given either fish skin (n = 16) or a hydrocolloid adhesive bandage (control) seven days after the burn. At days 7, 16, and 28 following burns, mice underwent histologic, hematologic, molecular, and gross examinations. When compared to hydrocolloids, the fish skin had no advantages for the overall closure of wounds. Furthermore, we did not find any differences in hematologic parameters or hypermetabolism between the fish skin and control treatments. In contrast to controls, the fish skin groups showed twice as much vascularization and twice as much antimicrobial defensin peptide expression. Antimicrobial peptides were found in the fish skin after proteomic profiling. Taken together, our findings imply that fish skin may offer burn patients a novel and affordable therapeutic option to promote vascularization and lessen bacterial infection.
Brazil is among the 180,000 people who die from burns every year, most of whom occur in low- and middle-income nations, according to the World Health Organization. In addition, non-fatal burns are responsible for extended hospital stays, incapacity, and ugliness, as well as rejection and shame. It has been proposed that Nile Tilapia Fish Skin (NTFS) be used as a biological material alternative for burn treatment. The presence of a normal, non-infectious microbiota was shown by the Colony Forming Units discovered in NTFS samples before the chemical sterilization treatment.
Burns can be divided into different categories based on how much harm they do. First-degree burns are classified as superficial burns that only cause harm to the skin. After an injury, these burns typically recover in seven days. The epidermis and dermis are affected by second-degree burns, which can be further divided into deep partial thickness (reticular dermis and epidermis) and superficial partial thickness (epidermis and papillary dermis). Superficial partial thickness burns usually heal in ten days and need daily wound care in addition to skin debridement. In addition to daily wound care, deep partial-thickness burns require surgical excision and resurfacing, with a more erratic healing duration of 10 to 28 days. A third-degree burn affects the entire dermis; it usually cures in three weeks or less and requires surgical excision and resurfacing. Burn injuries can cause systemic consequences in addition to merely skin burns. Burns that cover more than 30% of the body's surface area have the potential to cause a systemic inflammatory response, which increases the risk of sepsis, hypovolemic burn shock, and hypercoagulability. Moreover, hypermetabolism syndrome is brought on by the substantial metabolic challenge connected to severe burns. Even though the precise pathophysiology is yet unknown, it has been identified as a large rise in cortisol and catecholamines as well as modifications to acute phase protein pathways that result in elevated metabolic rates, systemic increases in catabolism, and multi-organ failure. Since its initial usage in a human clinical trial in Brazil in 2017, tilapia skin has been investigated as a potential burn therapy. Hu and colleagues reported that 58% of the hydrolyzed peptides isolated from tilapia skin had hydrophilic residues, while 99.14% of the peptides were smaller than 5 kDa. These peptides functioned well as an epidermal growth factor and reduced the scratch defects of keratinocytes. Additionally, compared to burn treatment and untreated control groups, the rabbits in the Vivo model treated with marine peptides had a higher wound-healing rate. The low cost of tilapia skin along with its availability and very simple farming makes it a desirable option for burn therapy. In addition, both wild and tamed animals suffered severe burn injuries as a result of recent wildfires in California. These animals' significant third-degree burn wounds were treated with tilapia skin because there were no reasonably priced skin replacement choices available. Clinical observations revealed a significant decrease in pain and a rise in comfort levels in tilapia skin-treated animals, despite the lack of controlled investigations. This study was motivated by clinical observations and aimed to investigate the effectiveness of tilapia fish skin bandages for full-thickness burns in mice as compared to controls treated with hydrocolloid bandages. Here, we present an analysis of the differences in wound sizes, serum glucose levels, body weights, complete blood counts, histopathology, and several molecular markers across the experimental groups. Furthermore, we present proteome analysis of the nonstructural peptides found in the skin of fish and mice.
Supplies and Procedures
- Research Plan
On the dorsum at day zero, scald burns developed in all of the mice. Body weights were noted, and serum and heparin tubes were filled with drawn blood. After the necrotic skin was surgically removed on the seventh day following the induction of burns, the exposed incision was wrapped with fish skin, hydrocolloid bandage (HCB), or adhesive bandage (control). In the fish-skin-treated and HCB controls, half of the mice were humanely killed on day 16, and the other half were killed on day 28 after the burns were inflicted. Similar to earlier, blood was drawn before to euthanasia. Following the animals' euthanasia, the fish skin and HCB bandages were taken off, the wounds were documented, and the animals' body weights were noted. An induced scald burn occurred on the dorsum. After the burned tissue was surgically removed on day seven, the wound was covered with either rehydrated fish skin or a hydrocolloid bandage (HCB). Metal clips held both bandages to the skin that remained undamaged. Animals were humanely put to death to collect tissue on days 16 and 28. On days 0, 7, 16, and 28, the infraorbital sinus was numbed while under anesthesia.
- The Debridement and Therapy of Burns
The mice were measured, unconscious, and had blood drawn using the same protocol on the seventh day after the burn. After the collection of blood, each mouse's injured location was surgically debrided, and the skin of fish or HCB (control) was used to fully cover the area of the wound.
- Preparing Fish Skins
With a pincher tool, the skin was removed from the very fresh tilapia fish (less than two hours after death) by pulling from the rostral-dorsal aspect of the fish's body towards the tail. To keep the excised skin from deteriorating while processing other fish, it was submerged in an ice bath for ten minutes. After that, the myofascial tissue was carefully scraped away using a firm scraper or a blunt knife. The skins were then cleaned in a fresh container with sterile 0.9% saline and placed in a container with 2% Chlorhexidine gluconate (VetOne 30159, VetOne, Boise, ID, USA), with one milliliter of solution for every square centimeter of skin. The skins had a 30-minute soak in this solution with mild stirring, followed by another saline rinse. The skins were soaked for one hour at room temperature with mild agitation in a 50% glycerol solution after undergoing the Chlorhexidine gluconate soak and rinse procedure once more. Then, for one hour at ambient temperature and then for twenty-four hours at four degrees Celsius, the skins were immersed in 75% and 99% glycerol (Thermo 17904, Thermo Fisher Scientific, Waltham, MA, USA), respectively. Following that, the skins were kept for a maximum of three months in 100% glycerol with 1% pen strep. In three consecutive soaks in sterile 0.9% saline, each lasting 20 minutes, the skins were rehydrated before being applied to the wounds. The full set of skins was used after they were rehydrated.
- PCR Examination
TRIzol (Invitrogen, Waltham, MA, USA) was used to extract total RNA from mouse skin by the manufacturer's instructions, and the QuantiTect Reverse Transcription Kit (Qiagen, Venlo, The Netherlands) was used to create cDNA. Using Qiagen's QuantiNova SYBR Green PCR buffer, real-time quantitative RT-PCR (Qiagen, Venlo, The Netherlands) was used to assess the RNA expression of the genes mentioned. Every reaction was tested in two sets. The cycle consisted of two minutes of denaturation at 95 °C, forty cycles of denaturation at 95 °C for five seconds, and ten seconds of annealing/extension at 60 °C. By Bio-Rad's CFX 96 (Bio-Rad Laboratories, Hercules, CA, USA), the last melting curve step was carried out. The target gene's expression level was calibrated against GAPDH. The delta-delta-CT approach was used to formulate all of the calculations. Primer sets were created using previously released data or known mouse gene sequences.
Samples of skin were digested tryptic in solution and then examined. As previously documented, proteomics samples were gathered, prepared, and subjected to LCMS analysis utilizing nanoAcquity (Waters, Milford, MA, USA) and Impact II (Bruker Daltonics, Billerica, MA, USA). To identify proteins present in samples, the sequences of identified peptides were searched against Uniprot proteomes (UP000005207, UP000000589) using PEAKS X Plus (BSI, Inc., Waterloo, ON, Canada) against mice and tilapia. The six-digit accession number of the identified proteins was then used to blast them against the NCBI. Researchers analyzed and filtered the peptides found in fish and murine skin, made a separate list of peptides found in fish skin that were absent in murine skin, and eliminated structural peptides found in both species' skin. The list was narrowed to uncharacterized nonstructural peptides after structural or identical peptides from both species were removed. Then, their sequences were thoroughly examined using the database of mammalian proteins. Here, using STRING Consortium Version 11.5 (STRING, Zurich, Switzerland), only proteins with greater than 90% similarity to unidentified peptides in fish skin are reported.
This is the first study to thoroughly and carefully assess, in a mouse model, the consequences of treating fish skin burns using histological, hematological, and molecular techniques. Additionally, we compared the proteomes of fish and mammalian skin, finding distinct proteins in fish skin that have anti-inflammatory, antibacterial, and wound-repair capabilities. When it came to wound closure, burn treatment with fish skin had no advantages over hydrocolloid treatment. There was no discernible variation in hematological parameters or systemic glucose levels between fish skin and HCB treatment. Nonetheless, compared to the HCB-treated controls, we observed that mice treated with fish skin showed more angiogenesis at the earlier time point. Additionally, we observed a trend of increased antimicrobial peptide expression in mice treated with fish skin as opposed to controls. When taken as a whole, these results point to the primary motivations for fish skin therapy for burns and suggest future lines of inquiry.
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