Evaluation of the Wound Healing Potential of Cynara humilis Extracts in the Treatment of Skin Burns

Cynara humilis is traditionally used to treat skin burns and microbial infections. However, experimental studies on this plant are rare. Furthermore, the aim of this study was to investigate the effects of Cynara humilis, a Moroccan herbal remedy, on the healing of deep second-degree burns in rats with a silver sulfadiazine group. This research was also carried out to confirm if C. humilis had antibacterial capabilities. Under typical burn procedures, each rat received a deep second-degree burn on the upper back. The burns were treated regularly with control groups (control and control VH), silver sulfadiazine (SDD) in group 3, C. humilis ethanolic extract (CHEE) in group 4, and C. humilis aqueous extract (CHAE) in group 5. Throughout the treatment, digital photography was used to measure rat responses to the treatment until day 18. After the scar biopsy at the end of the study, histological parameters (inflammatory cells, collagen, epithelialization, fibrosis, and granulation tissue) were assessed. Using the well technique, the antibacterial activity of the extracts was tested against Staphylococcus aureus CIP 483, Bacillus subtilis CIP 5262, Escherichia coli CIP 53126, Pseudomonas aeruginosa CIP 82118, and Salmonella enterica CIP 8039, and the results showed important activities of the ethanolic and aqueous extracts against the five species tested with MICs of 2 and 4 mg/mL, respectively. In the aqueous extract group, the wound healed faster. In addition, the healing rate in the C. humilis extracts (CHEA and CHEE) group was faster than in the silver sulfadiazine and control groups. In the C. humilis group, maximum wound surface recovery was observed at the same time, as it was not noted in the silver sulfadiazine group. Pathologically, epithelialization was more marked in wounds treated with C. humilis extracts (CHE). Angiogenesis and inflammatory cells were considerably lower in the CHE group than in the silver and other control groups. However, elastic fibers were considerable in the CHE-treated group. In histological examination, the C. humilis group had a low incidence of angiogenesis and inflammation, indicating that this group had less wound scarring. Collagen and burn wound healing were both faster in the C. humilis group. The findings of this study suggest that C. humilis, as indicated by traditional medicine, is a promising natural source for the management of wound healing.


Introduction
Burns are a global public health problem. Indeed, every year, millions of people sufer from skin burns all over the world [1]. Given the importance of skin as a barrier structure protecting the organism from external shocks, particularly infectious ones, postburn sequelae are one of the most problematic disorders in the medical world [2].
Over 90% of fatal burns from open fres occur in developing countries, with the poorest and middle-income populations bearing the brunt of the toll [3]. Burn-related mortality is estimated at approximately 265,000 deaths per year worldwide [4]. Te medicinal plants and native trees have been used in burn treatments and other natural remedies such as astringent properties and antibacterial efects [5].
Te creation of medicines that efciently and quickly heal damaged tissues while reducing treatment expenses greatly improves patients' quality of life. Phytotherapy, which uses the extract of diferent parts of plants, is one of the therapies employed. Nature has given medicinal agents, and an incredible number of contemporary medications have been identifed from natural sources, based on their use in traditional medicine [6].
Certainly, the wound healing efect of medicinal plants is due to their diverse bioactive components such as terpenoids, favonoids, tannins, and phenols. Herbal medicines had other advantages, including easy accessibility, fewer side efects, and low cost [7].
Decades ago, humans have used various portions of plants in the treatment and prevention of numerous illnesses since the dawn of time [8][9][10][11]. Opium, Physostigma venenosum (Eserine), cinchona bark, and cocaine are examples of herbal medicines that have been historically important for centuries [12].
Terapeutic medicine arose as a result of scientifc advances, and the notion of relying on traditional treatment began to fade. However, in many rural areas, the lack of hospitals or healthcare units, the high cost of pharmaceuticals, and the insufciency of modern treatments to effectively cure some human diseases have forced the rural population to rely on traditional and herbal medicines for their healthcare [5,13].
Te hunt for the antibacterial activity of natural extracts has intensifed. Natural biocides derived from higher and aromatic plants have long been used to control bacterial and fungal pathogens that cause disease in both humans and plants [14].
Antibiotics have revolutionized human health by allowing the treatment of life-threatening infections [15]. However, with the rise in bacterial resistance to currently available antibiotics, it is now more important than ever to hunt for innovative medicines derived from natural sources [16].
Medicinal herbs are widely used in Morocco, despite the lack of data in the literature. Indeed, the few local ethnopharmacological studies on medicinal plants used to treat skin burns showed 36 medicinal plants that are utilized to cure skin burns in various Moroccan localities [17][18][19].
Te current study focused on the Moroccan medicinal plant, namely Cynara humilis belonging to the Asteraceae family. C. humilis is very known by wild thistle, native to the Mediterranean region and traditionally used as a coagulant in the artisanal production of sheep's and goat's cheeses, and against burns for the healing process [17][18][19][20]. Ethnomedicinal surveys of C. humilis have indicated that the powder is most used by locals in Morocco. It is used as an anti-injury material [19], astringent, wound healing, and anti-infective agents.
However, to the best of our knowledge, the antibacterial activities of C. humilis roots have been examined in only one recent study [21]. Te aim of this study was to determine the wound healing and antibacterial efects and to evaluate the treatment of skin burns with C. humilis extracts.

Plant Collection and
Extraction. Te C. humilis roots ( Figure 1) were collected from the Skhirat region (Morocco) on April 2018. Te medicinal plant C. humilis was selected based on ethnopharmacological information from traditional herbalists in the region. Te identifcation of C. humilis was carried out, and the voucher specimen was deposited in the herbarium of the Scientifc Institute of Rabat in Morocco according to the code (RAB79161).
Te powder (250 g) of C. humilis root was macerated in one liter of distilled water under magnetic stirring for 24 h, at room temperature, and protected from light. Te extracts were fltered on Whatman paper and then concentrated under reduced pressure in the rotavapor at 50°C.
Te residue obtained was freeze-dried to remove all traces of solvent, and stored at +4°C. Te total aqueous extract (CHAE) and the ethanolic extract (CHEE) were prepared in the same way.

A Percutaneous Salve Preparation.
Te plant extracts in this study were used as a 20% ointment. Tey were combined with inactive ingredients that function as a moistureretaining patch, enhancing skin hydration and facilitating the absorption of active components. Te extracts were dissolved in purifed water and thoroughly mixed. Afterwards, in a mortar preheated in a water bath at 70°C and with the help of a pestle, lanolin was triturated with each prepared extract solution and added in small fractions until their total absorption. Ten, Vaseline was carefully incorporated into each mixture until a homogeneous ointment was obtained ( Table 1). Te prepared ointments were stored in sterile bottles in the refrigerator at +4°C.

Experimental Animals.
For our study, a total of 30 Wistar rats composed of male and female adults, weighing between 180 g and 300 g were used for dermal toxicity. A total of 30 adult male Wistar rats, weighing between 180 g and 250 g, were utilized for the wound healing investigation. Te female rats were nulliparous and not pregnant, and all the animals had healthy and intact skin.
Te animals used in this study were housed in cages and acclimated at a temperature of 22°C (±3), at a relative humidity between 30% and 70%, and subjected to a light/dark cycle of 12/12 hours. Te revival of the strains was done by adding to 100 μL strains preserved in glycerol at +4°C, 10 mL of tryptone soy broth TSB (tryptic soy broth), and incubation of the media for 18 hours at 37°C. For the minimum inhibitory concentration (MIC), a modifed resazurin microtitre-plate assay was used as reported by Sarker et al. [22] and Marmouzi et al. [23].

Antimicrobial
Under aseptic conditions, serial dilution was achieved by transferring 100 μL of the test material from the frst row to successive wells of the next row of the same column, then dosing 100 μL of the material at sequential drop concentrations into each well.
After adding 10 μL of a bacterial suspension (10 8 CFU/ mL) to each well, the microplates were incubated at 37°C for 24 hours. A column containing an antibiotic was used as a positive control (chloramphenicol in sequential dilution at a concentration of 25 mg/mL). For all plant extracts in suspension, the concentrations used were 32 mg/mL. Finally, a resazurin solution was prepared by dissolving 270 mg of resazurin powder in 40 mL of sterile distilled water. Te solution was stirred until the powder was completely dissolved and the solution became homogeneous [24]. After incubation, a volume of 10 μL of resazurin solution (5 μm/mL) was added to each well. Ten, the microplates were placed in a 37°C incubator for only 2 hours. Te color shift was then visually examined. Any color changes from purple to pink or colorlessness were considered positive. Te MIC values were determined by taking the lowest concentrations at which color change occurred.

Dermal
Toxicity. Acute dermal toxicity of C. humilis extracts was performed on 30 Wistar rats comprising males and females according to the OECD 402 limit test (OCDE, 1987) at a dose of 5000 mg/kg body weight. 24 hours before the test, using an electric tensioner, the dorsal region of the trunk of the animals was shaved, in which this area presented 10% or more of the total body surface in order to avoid skin lesions. Te animals were divided into three groups of 10 rats each, with an equal number of males and females (n � 10).
Te control group was treated with the vehicle (water), and the test groups were treated with the aqueous and ethanolic extracts. Te previously weighed dose was moistened with water and then applied to the shaved part of the animal by performing a slow massage to increase the penetration of the product. Te area was then covered with gauze and held in place with a bandage for 24 hours. Ten, the dressing and the gauze were removed, and the skin was cleaned with lukewarm water and then each animal was returned to its individual cage.
After substance application, the rats were observed every 30 minutes for 10 hours on the frst day and daily for 14 days. During this observation period, the weight of each animal was measured once a week, and the behavioral manifestations, the signs of local and systemic toxic symptoms, and the number of deaths were recorded daily.
The C. humilis plant The root of C. humilis  Evidence-Based Complementary and Alternative Medicine 2.6. Wound Healing Properties. Te rats were anesthetized by injecting 100 mg/kg ketamine hydrochloride intraperitoneally, and their backs were shaved using an electric clipper and disinfected with 70% ethanol. A deep-thickness second-degree skin burn on the back of the rats corresponding to a 3.14 cm 2 was performed based on earlier studies with minor modifcations [25]. Te rats were randomly divided into fve groups of six animals in each group as follows: Control group: animals induced with a wound not treated with any drug VH control group: animals treated with the excipients of the containing ointment (purifed water, petroleum jelly, and lanolin) SSD Group: animals treated with the standard silver sulfadiazine 1% ointment CHEE group: animals treated with an ointment containing 20% of the ethanolic extract of C. humilis CHEE group: animals treated with an ointment containing 20% of the aqueous extract of C. humilis Wound treatment was performed immediately after induction of the burn and once a day until the 18th day. Approximately, 0.5 g of each treatment was evenly applied to the wound to cover the entire burned area. Te burned animals have been caged individually.

Retraction Evolution Protocol.
Te clinical evaluation of the animals has been conducted by monitoring their weight every three days. Te clinical evolution of the wounds was carried out daily in the morning by macroscopic examination of the lesions throughout the healing period of the burns. Tis examination is based on the observation of the appearance of changes in the color, odor, and appearance of the wounds, the formation of edema, bubbles, or crusts, bleeding, swelling, and secretion.
Wound healing was assessed by calculating the rate of wound retraction. For this purpose, the wound surfaces were measured at D 6 , D 12 , and D 18 days after burn induction.
Te burn site was photographed using a Canon SX710 HS digital camera (20.3 megapixels; zoom � 1). In parallel, a direct manual tracing of the wound surfaces on transparencies was carried out. Te photos and transparencies were analyzed by image processing software (ImageJ software) to calculate the surfaces and determine the retraction rate. Te wound retraction rate was calculated by the following formula according to Senthil Kumar et al. [5].
Percent wound retraction of J � Initial wound size J0 − specific day wound size Jx Initial wound size J0 × 100. (1)

Histological Analysis.
In this study, light microscopy was used to examine the histology of the skin. Te histological evaluation was carried out at the end of the experiment on day 18 after the sacrifce of all the animals under ethical conditions. Te biopsies were performed by taking a sample from the scar using a scalpel. Fixation was performed with 10% neutral bufered formalin (NBF) for 24 h at 4°C. After being dehydrated in alcohol, the skin samples were embedded in parafn wax (58-60 mp). 4 µm thick skin sections were stained with hematoxylin, green trichrome from Masson, and orcein. Stained skin slices were shot with a camera linked to a photo microscope. Te degree of infammatory response, collagen and elastic fber deposition, angiogenesis, granulation tissue development, and epithelialization were all measured histologically to determine wound healing capacity. Each parameter was given a score ranging from 0 to 3 [26]. Masson's trichrome green staining and orcein staining were used for qualitative estimation of collagen deposition and elastic fbers, respectively.

Statistical Analysis.
Te statistical analysis was performed using RStudio 4.6 software, and data were expressed as the mean ± standard deviation (SD) for each measurement. Te data were also analyzed by one-way analysis of variance (one-way ANOVA). Posthoc procedure was used for the signifcance of diference (p < 0.05).

Antibacterial Activity.
In our study, the antibacterial activity of two extracts from the roots of C. humilis was studied against fve bacterial species. Te results ( Table 2) showed that the ethanolic extract possesses antibacterial activities against S. aureus CIP 483, E. coli CIP 53126, B. subtilis CIP 5262, S. enterica CIP 8039, and P. aeroginosa CIP 82118 at a similar minimum inhibitory concentration (MIC) equal to 2 mg/mL for all bacterial species. Te aqueous extract also revealed antibacterial activity against the fve bacterial species at MICs equal to 4 mg/mL but lower than those of the ethanolic extract.

Acute Dermal Toxicity.
From day 0 to day 14, the body weights of both the control and treatment groups were gradually increased, as shown in Figure 2. Tere was no signifcant diference in average weight between the two groups. During the 48-hour observation period, no meaningful clinical symptoms of C. humulis toxicity at a dosage of 5 g/kg were noticed. On the other hand, the animals stayed normal for the next 14 days and no death was observed. Figure 3 represented the evolution of the average body weights of the rats receiving the diferent treatments every three days. Te mean body weight of rats in the control VH and SSD groups was not statistically diferent (p < 0.05) from the control group. Te gain in body weight during the experimental period (D 0 to D 18 ) was 3 g, 2.33 g, and 2.67 g, respectively, for control, control VH, and SSD.

Body Weight Evolution (in Burn Study).
Unlike to the two groups treated with plant extracts, the weight of the rats having received CHAE was increased from 184.17 g on day 0 to 200.5 g on day 18, representing a gain in body weight of 16.33 g. From 202.0 g (day 0) to 211.17 g (day 18) for the CHEE group rats, a body weight gain of 9.17 g was obtained.

Macroscopic Evaluation of Rat Wounds by Cynara humilis.
Topical use of C. humilis compounded into an ointment base on wound healing in rats resulted in a considerably (p < 0.05) greater rate of wound healing in rats in the current investigation. Te wound area after C. humulis ointment administration was indicated in Figure 4. Wistar rats treated within 12-18 days had a superior healing pattern with full wound closure (Figure 4), whereas the control group needed more than 18 days. Figures 5-7 demonstrated the diferences in wound area assessed on days 6, 12, and 18, after burn in the three treatment groups compared to the two control groups. Eighteen days after the burn, the extract EECHand EACH-treated groups had achieved more than 90% wound healing. Te CHAE-treated group had achieved a healing rate of 32% on day 6 and 71% wound healing after 12 days, which was not substantially diferent (p > 0.001) from the group treated with the CHEE extract. Te percentage of healing in the SSD-treated group was substantially lower (p < 0.001) than in the extract-treated group, with 65.7% on day 18. It can be seen that the wound contraction percentage of C. humilis ethanolic extracts (97.58%) and aqueous extract (94.31%) after 18 days is substantially higher (p < 0.001) compared to untreated group (64.2%) and the groups treated with vehicle control (75.3%).

Histological Results.
On the 18th day of treatment after induction of the burns, the histological evaluation of the wounds was carried out using three types of staining. Figures 8-10 shows histological sections of wound scars observed under an optical microscope. Te control, control VH, and SSD groups showed nonreepithelialized epidermis after 18 days of treatment. Te dermis showed ulceration with the presence of necrotic tissue debris mixed with polymorphic infammatory cells with congestion and edema ( Figure 8).
Te treatment of rats with C. humilis extracts resulted in signifcant epithelialization and a decrease in infammatory cells with the absence of necrotic tissue debris. Tere was also the production of a modest amount of extracellular matrix and the development of new blood vessels and hair follicles (Figure 8).

Evidence-Based Complementary and Alternative Medicine
Te study of collagen fber deposition and organization was conducted using Masson's Trichrome Vert staining for histological evaluation (Figure 9). Te results revealed low density and poor organization of collagen fbers in rats from the control, control VH, and SSD groups, along with persistent epidermal ulceration and a polymorphic infammatory infltrate in the dermis with edema, indicating an incomplete healing process (Figure 9). Te groups treated with C. humilis extracts showed an improvement in wound healing, as evidenced by a high density and better organization of collagen, as revealed by the analysis of collagen quantity and quality (Figure 9).
Te deposition of elastic fbers was analyzed after staining the histological sections with orcein. Under an optical microscope, the elastic fbers were visible as wavy bundles, sometimes connected, with a reddish-brown color on a beige background ( Figure 10). Te results showed the abundance of elastic fbers in the histological sections in the groups treated with C. humilis extracts, unlike the other groups.
Histological analysis of wound biopsies was also performed based on a scoring system by studying collagen and elastic fber deposition, infammatory response, appearance of granulation tissue, angiogenesis, and epithelialization (Figures 11-16).     Evidence-Based Complementary and Alternative Medicine In all groups, the histopathological picture is nearly identical. On the eighth day, the number of infammatory cells was higher in wounds of control, TVH control, and SSD groups than the groups treated with the creams containing 20% of the C. humilis extracts (Figure 9).
All groups displayed moderate new blood vessel formation (angiogenesis), with the control and TVH control groups exhibiting modest angiogenesis (Figure 12). Te proportion of collagen bundles in microscopic felds was used to score collagen synthesis in the skin. Collagen analysis revealed no signifcant diferences between C. humilis extracts (EECH and EACH) and a low score diference between control and TVH control ( Figure 13).
However, there is no signifcant diference in reepithelialization score between the control and TVH control (p > 0.05), while there is a signifcant diference when        Evidence-Based Complementary and Alternative Medicine comparing controls with SDA therapy (p > 0.05). On EACH and EECH, the greatest reepithelialization score ( Figure 14) was found, and it exhibited comparable signifcance (p < 0.01). Te treatment groups (EECH and EACH) had superior reepithelialization observations than the controls (control and TVH) and SSD groups (p < 0.05). Collagen quantity and quality were both raised in C. humilis extracts. Te plant therapy groups had improved infammation and angiogenesis processes. Te deposition of elastic fbers is highly controlled over the 18 days (Figure 15), and comparable patterns were detected across (control, TVH control, and SDA) with a low signifcant diference (p > 0.05). Low score of elastic fbers was observed in scars that were treated with SDA and control groups. During the 18th day, substantially more elastin fbers were found inside the dermal scar tissue in the two treatment groups that received EECH and EACH in wound healing after burn skin (p < 0.05). Te volume and distribution of granulation tissue on the group treatments ( Figure 16) were considerably lower in the C. humilis ointment group (p < 0.01) than in the control group (p > 0.05). Te best wound healing results are shown in EECH and EACH (C. humilis treatment), which have a high density of collagen with a good organization, a full and mature epithelium, a low number of infammatory cells, and angiogenesis, respectively.

Discussion
Burn wounds are particularly vulnerable to complications [27,28]. Complications can wreak havoc on the healing process in a variety of ways. An infection is the most likely consequence, according to this study. As a result, the comparison between standard drugs (chloramphenicol) and topical drugs (C. humilis extracts ointment) revealed that topical drugs were highly active and similar to chloramphenicol against Gram-negative and Grampositive bacteria, as well as Pseudomonas aeruginosa, were inexpensive, and had no harmful side efects [19,23]. Te current investigation found that an ethanolic extract of the root of C. humilis had potent growth inhibitory properties against multiresistant Gram-positive and Gramnegative bacteria. A very recent study investigated the antimicrobial activity of hydroethanolic extracts from C. humilis roots and leaves against fve pathogenic microbial strains (2 E. coli, 1 coagulase-negative Staphylococcus, Klebsiella pneumoniae, and Candida albicans) using two techniques: microdilution and agar disk difusion. Te results showed that the extracts had an efect against all tested strains, with inhibitory zone values ranging from 12 to 15 mm. Te root extract showed the strongest activity against coagulase-negative Staphylococcus (IC 50 � 6.25 mg/ mL) [21]. Other fndings given by Al-Rimawi et al. [29] looked at the antibacterial activity of methanol and ethanol extracts from P. halepensis Mill. Seeds, bark, and cones against S. aureus and E. coli. Although a pomegranate antimicrobial study reveals the antibacterial process, the active ingredient inside the pomegranate causes toxicity in bacteria by changing and interacting with enzymes and substrates [30]. In contrast, C. humilis has been shown to be rich in sesquiterpene lactones [31], which are known for their high antibacterial potential [32].
Te use of silver sulfadiazine as a control positive (SDA group) in this study did not produce the best results in terms of collagen creation; instead, it produced the worst results. Te unfavorable efects of silver sulfadiazine that are clearly evident in this experiment include wound contracture retardation and delayed and incomplete epithelialization, which are also reported in a work by Muller et al. [33]. Te risk of delayed wound healing is increased when the infammatory process is prolonged and severe. Skin layer renewal is one of the most signifcant indicators of burn wound healing during therapy [34]. According to our histological analysis, C. humilis extract therapy showed regenerative efects on the dermal layers. Recently, several studies have reported the discovery of natural compounds, including those derived from medicinal plants, for the creation of new burn wound healing drugs. In this context, our study attempted to assess the burn wound healing of an ointment made from a Moroccan medicinal plant that is well known for treating skin burns. Natural herbal materials are used in Asian and Indian medicine because they have advantages over manufactured medications. Te inclusion of numerous active ingredients in herbal medications may potentially broaden the therapeutic range and improve  Evidence-Based Complementary and Alternative Medicine treatment outcomes [35]. However, no research on the efcacy of C. humilis ointment against burn wounds has been performed, according to the literature. Our research looked at the healing of burn wounds in this example. Te impact of C. humilis extract on rats developing burn wounds was remarkable.
In the research conducted by Akhoondinasab et al. [36], the aloe vera group had a considerably shorter healing time in grade 2 wounds than the silver sulfadiazine group. Because contraction plays a large role in second-degree wounds, especially in rats' slack skin, this impact was more noticeable in the second-degree burn. In our situation, the contraction rate was greater in the EECH and EACH groups than the SDA and control groups. Furthermore, epithelialization is a critical component of wound healing and determines its success. Reepithelialization is required for a wound to be deemed healed. Histological data showed that all of the treated groups had a higher rate of epithelialization than the control group.
Infammation has three functions: clearing damaged cells, vasodilation, and infammatory cell extravasation. In our investigation, C. humilis ointment had a signifcant efect on collagen and infammatory cells. It was comparable to earlier fndings, which showed that applying avocado and jojoba oil topically to rats increased collagen production and reduced the number of infammatory cells during wound healing [37][38][39].
Infammation can also aid in the production and degeneration of dermal cells. According to these facts, burn wounds, which are subject to a high level of infammation and cannot be avoided, should be treated with antiinfammatory and antioxidant drugs to reduce the risk of delayed wound healing [7]. As proven in our previous work [13], C. humilis has a variety of active compounds, each of which can operate as an antibacterial, anti-infammatory, and powerful antioxidant.
Te deposition of collagen in the burn skin was higher in the EECH-treated group's wounds. Indeed, epithelialization was more prominent in the wounds treated with C. humilis extracts. Other honey treatment results from Sushma et al. [40] revealed that all treated groups had better collagen deposition at the wound site than the control group. Te collagen organization in the treated groups was ordered and horizontally interwoven. In contrast, the collagen fbers in the control group were arranged obliquely and loosely. Angiogenesis is one of the most researched processes in a burnt wound [41].
Te results of excellent elastic fbers and minimal angiogenesis production were associated with an efective burn wound healing process in this study. Collagen production is signifcantly boosted when the dosage of C. humilis extract is raised to 20% ointments. In comparison to control and standardization of silver sulfadiazine, this result shows that ointment generated from C. humilis extract with 20% ointments may establish an excellent process of burn wound healing in rats. According to these fndings, using a 20 percent ointment of EACH and EECH extract to assist an ideal burn wound healing process likely provides a synergic mechanism.
It is crucial to highlight that the extracts did not cause discomfort or pain to the rats throughout the brief duration of wound therapy, since the rats showed no symptoms of restlessness or scratching or biting of the wound site when the extracts were administered.

Conclusion
Te antibacterial activity of crude extracts of C. humilis as well as wound healing capabilities was verifed in this investigation. In the animals with acute dermal toxicity, there are no treatment-related signs of toxicity or death. Te extracts of these plants, on the other hand, might be turned into phytomedicines for the treatment of infected wounds. Te results demonstrated that EACH and EECH ointments are successful in treating second-degree burns in rats and may be suitable for short-term therapeutic treatment of second-degree burns. However, other investigations should be conducted to determine the chemical compounds of plant extracts as well as to investigate their antibacterial and wound healing properties. Moreover, other toxicological studies should be investigated to confrm the safety of plant extracts and their bioactive compounds.

Data Availability
Te data used to support the fndings of this study are included within the article.