Polyphenolic Profiling and Wound Healing Potential of Hammada scoparia (Pomel) Iljin: Preclinical Evaluation of a Novel Topical Formula

Abstract

Objective

Hammada Scoparia (Pomel) as an endemic plant from the Amaranthaceae family, holds a significant position in the Saharan region of southern Algeria's traditional medicine. The aerial parts of Hammada scoparia was used to cure inflammation and wound healing management. The aim of this study was to determine the phytochemical constituents of the aqueous extract obtained from the aerial parts of Hammada scoparia (AEHs)and assess its effectiveness in wound healing.

Methods

Formulation of Wound healing cream were prepared from Hammada scoparia aerial parts aqueous extract then the wounds were applied on the skin of the dorsal region of each Wistar albino for 13 days. Hematological, inflammation and histological parameters were evaluated. Different bioactive compounds were identified within AEHs using HPLC analysis.

Results

High-Performance Liquid Chromatography (HPLC) analysis identified 21 phenolic compounds, with rutin being the most abundant, followed by naringenin, gallic Acid, valinin, and hydroxy-comarin. The formulated AEHs cream exhibited superior wound healing properties, with a 96.54% healing rate by day 13, outperforming the DOUCE PLUS cream and control groups. Physical analyses indicated the cream's white color, neutral odor, humid texture, pH of 6.8, and excellent stability. Hematological parameters revealed the AEHs cream influenced red blood cells (RBCs), white blood cells (WBCs), and platelet (PLT) levels positively. Inflammation markers, including C-reactive protein (CRP) and sedimentation rate (ESR), were also favorably modulated by the treatment. Histological studies indicated that H. scoparia cream promoted re-epithelialization, fibroblast activity, collagen deposition, and angiogenesis, thus facilitating an effective healing process.

Conclusion

The study concludes that Hammada scoparia possess significant wound healing properties, corroborating their traditional medicinal use.

Introduction

The Wounds have affected humans since prehistoric times and the healing of wounds is an art as old as humanity. As life expectancy continues to increase, coupled with modern lifestyles, wounds—especially chronic wounds—have become more prevalent, current estimates suggest that nearly 6 million people suffer from chronic wounds. ¹ Wound healing involves four overlapping phases: hemostasis, inflammation, proliferation, and remodeling. ² Impaired local or systemic immunity can extend the inflammatory phase and delay healing. The healing rate is influenced by various factors, including wound size, blood supply to the affected area, presence of infection, and foreign objects. ³ Wound care may require the use of both local and systemic medications. Additionally, several growth factors, such as macrophage-derived, monocyte-derived, and platelet-derived growth factors, are essential for accelerating wound healing. ⁴ To suppress excessive inflammation and expedite the healing process, anti-inflammatory drugs are necessary to stimulate wound healing. ⁵

Numerous medicinal plants have demonstrated wound healing properties. Extensive research has been conducted on the role of medicinal plants in wound healing management.^6–11 Various plants and their preparations have been traditionally used for wound treatment due to their significant impact on the healing process. Examples include Vernonia amygdalina Del., ¹² Brucea antidysenterica J.F.Mill, ¹³ Achyranthes aspera, ¹⁴ Croton macrostachyus, ¹⁵ Datura stramonium, ¹⁶ Bersama abyssinica, ¹⁷ Rumex abyssinicus, ¹⁸ Cordia Africana, ¹⁹ Aloe spp., ²⁰ among others.

Hammada scoparia, an endemic plant of the Chenopodiaceae family, is widely regarded as a potential source of therapeutic drugs. ²¹ Known for its medicinal properties, it is frequently used in North African folk medicine to prevent various ailments, including hepatitis, inflammation, and obesity.^22,23 H. scoparia is a highly potential reservoir of bioactive compounds. Furthermore, its potential has been demonstrated in exhibiting antioxidant,^24,25 antimicrobial, ²⁵ and anti-inflammatory properties ²⁵ that significantly contribute to improving wound healing.

In Algeria, scientists assessed the intrinsic pharmacological characteristics of these plants by studying their antioxidant, ²⁶ antidiabetic, ²⁷ antimicrobial, ²⁵ and antileukemic capabilities. ²⁸ Previous studies have shown data that support the traditional uses of H. scorpia decoction, specifically the extract from its aerial parts, in treating diabetic disease, ²⁹ inflammations, and wounds, ³⁰ and mouth diseases and toothache. ³¹

Comprehensive phytochemical investigations on this plant have shown that the aerial parts of H. scoparia are abundant in flavonoids, including prominent compounds like Isorhamnetin 3-O-β-D-xylopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→6)-β-D-galactopyranoside and Chrysoeriol. ³² Furthermore, stems of H. scoparia contain considerable amounts of phenolic acids such as cinnamic, coumaric, catechic, and vanillic acids.^27,33 Phytochemical analysis of the H. scoparia flower in Algeria has detected the presence of flavonoids, tannins, terpenoids, and alkaloids. These compounds, due to their antioxidant and anti-inflammatory properties, have the potential to improve the wound healing capabilities of H. scoparia significantly.

Despite numerous traditional claims on the wound healing properties of Hammada scoparia (Pomel) Iljin. Nevertheless, so far, no relevant literature evidence has been discovered regarding their wound-healing properties. Therefore, the results of this study could provide baseline information for future research. The aim of this study was to evaluate the wound healing activities of crude extract of Hammada scoparia (Pomel) Iljin arial parts.

Material and Methods

Plant Material

Hammada scoparia (Pomel) Iljin, also known as Remth, in the South of Algeria (Biskra), has been documented as a medicinal plant at the World Flora Online (WFO) https://www.worldfloraonline.org/taxon/wfo-0000715323). The aerial parts of H. scoparia (Pomel) Iljin were collected in November 2021 in Southeastern Algeria, specifically in the province of Biskra (34° 51′ 00″ North, 5° 44′ 00″ East). The plant material was identified by Professor Hammel Tarek from the University of Badji Mokhtar - Annaba, Algeria. After collection and transport, the aerial parts were washed with running water to remove dust, dried in the shade at room temperature, powdered, and stored for further use. Under the code 202111Bis/HAMsc, a plant specimen has been deposited to the Laboratory of Biology, Environment and Health (LBEH) herbarium.

Animals

A total of 20 adult male albino rats, weighing 160–200 g, were obtained from Pasteur Institute, Algeria. They were placed and kept in the animal house of the Molecular and Cellular Biology Department, University of El-Oued, Algeria. Animals were adapted for 15 days under the same laboratory conditions of photoperiod (12 h light/12 h dark) with relative humidity and room temperature (22 °C) of standard rats. Food and tap water were available for the duration of the experiments. ³⁴

Aqueous Extract Preparation

Fifty grams of powdered H. Scoparia was soaked in 500 mL of distilled water over one night at ambient temperature. Whatman paper N°1 was used to filter the mixture, and the residues were macerated for another two days with the same solvent and volume. The filtrates were collected and concentrated with rotatory vapor (Buchi R-300H) under low pressure and a temperature of 40 °C. The yield was calculated, and the extract was stored for further studies. ³⁵ The extraction yield was assessed using the method mentioned by Muniyandi et al (2019). ³⁶ It was calculated using the following formula:

$$\mathrm{Extraction}\mathrm{Yield}(\text{\%})=({\mathrm{M}}_1/{\mathrm{M}}_0)\mathrm{x}100,$$

Where M₁ represents the mass in grams of the resulting dry extract, and M₀ represents the mass in grams of the plant material processed.

HPLC Analysis

Reverse-phase high-performance liquid chromatography and scanning instruments were used to identify the active components. RP-HPLC with a UV-Visible detector (Shimadzu LC20 AL) was used to investigate the presence of phenolic content in the crude extract. Shim-pack VPODSC18 (4.6 mm, 250 mm, 5 m) analytical column, UV-Vis's detector SPD 20 A, and universal injector (Hamilton 25 l) were all incorporated into the system (Shimadzu). Non-polar aliphatic residues were used in the RP-HPLC experiments, and the mobile phase was made up of a gradient elution of acetonitrile and acetic acid (0.1%). The flow rate employed in this study was 1 mL/min, and the injection volume was 450 mL. The sample and standard injection volumes were 20 mL, and the monitoring wavelength was 268 nm. Various substances were determined by comparing their UV absorption and retention time to standards. The identification of the phenolic compounds was based on comparing the retention times and UV absorption spectra of the sample peaks with those of known standards. This method ensures reproducibility, as the UV absorption spectra and retention times for each compound are consistent when using the same experimental conditions.

Wound Healing Activity

Formulation of Wound Healing Cream

This study aimed to enhance the formulation of a water-in-oil type healing cream, composed primarily of two phases: a fatty phase consisting of sweet almond oil, and an aqueous phase containing distilled water and H. scoparia extract. Emulsification between these phases was achieved using E-wax and Glyceryl stearate with peg 100. Additionally, Vitamin E was incorporated as a preservative. Regarding storage conditions, it recommend storing the cream in a cool, dry place, away from direct sunlight, based on the nature of its ingredients.

Physical Analyzes of Formulated Cream

pH Test

The pH measurement of the cream is measured by diluting a quantity of the samples with water, then filtered with Watman N°4 filter paper. ³⁷ The pH analysis is carried out at a temperature of 20 °C. using a pH meter (HANNA-HI 8424).

Stability Test

This test was carried out on the sample after centrifugation at 3000 rpm, for 30 min and at room temperature. The degree of separation of the two phases is expressed by the total percentage of stability, ie (100 ═ stable, 0 ═ unstable). ³⁷

The cream demonstrated a shelf life of more than 12 months, which indicates its stability over an extended period. This was confirmed through preliminary stability tests, where the cream maintained its consistency, pH, and effectiveness without any signs of phase separation or ingredient degradation during this period.

Experimental Design

Wistar albino rats served as an experimental model for creating wounds. The rats came from Institute Pasteur, and the experiment was carried out in the laboratory of the molecular and cellular biology department. The animals were divided into seven groups, each comprising 05 rats: Group I: The rats were not exposed to any wound induction and did not receive any cream treatment for 13 days; Group II: The rats were exposed to wound induction and left without cream treatment; Group III: The rats were exposed to wound induction, and received Hammad scoparia cream treatment for 13 days; Group IV: The rats were exposed to wound induction, and received DOUCE PLUS^® cream treatment for 13 days.

Induction of Wounds

The wounds were made on the skin of the dorsal region of each animal; the choice of this region is motivated by ease of access, both for making the wound and for monitoring the progress of healing and taking measurements. To create wounds in rats, the animals were first anesthetized using an appropriate anesthetic (xylazine), and their skin was shaved where the wound would be induced and disinfected with 70° surgical alcohol. The wound area was marked with a sterile surgical marker to ensure consistent dimensions across all animals. For circular wounds, a sterile biopsy punch (10 mm) was used to create a uniform wound size, making sure not to cut deeper than the dermis to maintain consistent depth. After the excision, the wound was cleaned with saline, an antiseptic ointment was applied, and post-surgical analgesia (buprenorphine) was provided. Just after wound induction, the animals in the treated batches each received a topical application of the product intended for their respective batch. Treatments were applied once every 24 h for each rat. Treated or untreated wounds were not protected by a dressing. The wound size was consistently measured and documented, with standardized photographs taken, and the wounds were monitored daily for signs of healing or infection.

Evaluation of Cicatrisation Process

After photographing them, the dimensions (length, width) of the excision wounds are measured every 3 days during the trial period (13 days). The percentage of the evolution of wound cicatrisation is calculated using the following formula ³⁸ :

$$\begin{array}{r}\mathrm{Wound}\mathrm{Healing}(\text{\%})\\ =(\mathrm{cicatrized}\mathrm{wound}\mathrm{surface}/\mathrm{initial}\mathrm{wound}\mathrm{surface})\times 100\end{array}$$

Sacrifice and Blood Collection

After a 16-h fasting period and completion of each treatment, rats were euthanized under light anesthesia with 94% chloroform. Blood samples were collected into pre-marked and numbered EDTA tubes for each rat. The blood was then centrifuged at 2500 rpm for 10 min, and the resulting serum was stored at −20 °C until biochemical analysis.

Histological Study

Skin tissue samples were obtained from the wound sites of all Wistar albino rats to create histological sections for monitoring the stages of wound healing. The skin fragments previously fixed in 10% formalin are placed in cassettes which are then placed in an automated machine. The organ fragments are first dehydrated by submersion in ethanol baths at increasing concentrations (60%, 70%, 80%, and 100%). The samples undergo two baths of xylene and two others of melted paraffin. Xylene occupies the place of water and therefore facilitates the penetration of paraffin since the latter is hydrophobic. The duration of the baths is 24 h. Using an embedding device, the skin samples are placed in metal molds and covered with melted paraffin. After cooling, the blocks are ready for cutting. The blocks are placed in the microtome to make sections 3 μm thick. Using very fine forceps, the sections are placed on slides which are then deparaffinized by heating in an oven for one hour. To highlight the layers of the skin, the sections are first rehydrated by successive submersion in the following baths: xylene bath (5 min), ethanol bath (5 min). After rinsing in distilled water (5 min), the rehydrated sections are placed in a hematoxylin bath (5 to 6 min). The excess dye is removed by a water bath with a few drops of ammonium hydroxide (NH ₄OH), then they are put in an eosin bath (5 min) to color the cytoplasm and the excess dye. dye is removed by ethanol. The slides thus stained are covered with lamellae and are suitable for microscopic observation.

Statistical Analysis

The results are presented as the mean ± standard error of the mean (Mean ± SEM). Group comparisons were conducted using one-way ANOVA. Statistical analysis was performed using EXCEL (Version 2021). Significant differences between means were determined using Duncan's multiple range test (DMRT) at a significant level of P = .05 with SPSS (Statistical Package for Social Sciences) version 29 software. Differences were considered significant at P ≤ .05.

Results and Discussion

HPLC Analysis

The chromatogram profile of the aqueous extract unveiled the presence of 21 phenolic compounds (Table 1 and Figure 1). By evaluating retention times and peak areas, the nature of these compounds was discerned. Rutin emerged as the most abundant, constituting a major peak at 28.288%, succeeded by Naringenin (13.662%), Gallic Acid (11.037%), Valinin (9.436%), and Hydroxy-Comarin (6.006%). Among the less dominant compounds, including Beta-Carotene, Ferulic Acid, Salicylic Acid, Caffeic Acid, and ascorbic acid, their quantitative significance varied between 2.8% and 3.7%. Ferulic acid, myricetin, and quercetin exhibited lower prominence, with contents ranging from 0.785% to 2.101%, these compounds possess notable biological activities, particularly in terms of their antioxidant, antiseptic, analgesic, and anti-inflammatory properties, potentially rivaling those detected at higher concentrations.

Figure 1.

HPLC/UV chromatogram profile of the aqueous extract of Hammada scoparia.

Figure 2.

Time schedule of the experiment in vivo wound healing activity of different cream treatment formulations; n = 5.

Figure 3.

Wound healing rate at day 4, 7, 10, and 13 in vivo treatment of different cream treatment formulations; n = 5.

Table 1.

HPLC Results of the Aqueous Extract of Hammada scoparia.

N°	Name	Ret. Time	Height	%Area
1	Lawsone	0.169	1852.000	0.675
2	Keampferol	0.228	1868.000	0.683
3	Ascorbic acid	0.943	2920.000	2.101
4	Coumaric acid	0.946	4662.000	0.803
5	Rutin	4.499	601,206.000	28.288
6	Gallic acid	8.147	4601.000	11.037
7	Caffeic acid	11.823	5890.000	1.899
8	Chrysin	12.539	709.0000	0.540
9	Maleic acid	17.329	8015.000	0.310
10	Ferulic acid	19.112	754.0000	0.981
11	Naringenin	21.614	6421.000	13.662
12	Quercetin	22.786	631.0000	0.612
13	Viniferin	31.233	34.000	0.012
14	Chlorogenique acid	32.428	85.000	0.016
15	Beta Carotene	32.579	2444.000	0.785
16	Hydroxy coumarin	37.719	9757.000	6.006
17	Folic acid	40.017	0.000	0.000
18	Thymol	40.989	0.000	0.000
19	Hepertin	43.159	174.000	0.650
20	Salicylic acid	47.261	1723.000	1.922
21	Valinin	47.553	16,560.000	9.436

The HPLC analysis of the aqueous extract of Hammada scoparia reveals significant differences in phenolic compound composition between this study and that of Benkhrara et al (2021). ²⁷ In this study, the most prominent compounds by %Area were rutin (28.288%), gallic acid (11.037%), and naringenin (13.662%). Minor compounds included ascorbic acid (2.101%), coumaric acid (0.803%), and caffeic acid (1.899%). Comparatively, benkhrara et al (2021) identified ascorbic acid (2.8%), gallic acid (24.5%), and caffeic acid (2.4%) among others. Notable compounds from their study included resorcinol (4.2%), catechin (3.1%), gentisic acid (3.4%), and epicatechin(2.0%). ²⁷

Physical Analyzes of Formulated Cream

The results concerning the Physical analyses of formulated cream used in this study are mentioned in Table 2. The pH analysis is carried out at a temperature of 25 °C using a pH meter (HANNA-HI 8424). The degree of separation of the two phases is expressed by the total percentage of stability, ie (100 ═ stable, 0 ═ unstable).

Table 2.

Physical Analyzes of Formulated Cream.

Color	Odor	Texture	pH	Stability
White	No	Humid	6.8	100

The results represented in Table 2 offer insights into the properties of a skin cream formulation which is described as white appears neutral. Its lack of odor suggests it is formulated without added fragrances, making it potentially suitable for individuals with sensitivities to scented products. The texture is labelled as “humid,” indicating a moisture-rich consistency that may provide hydration and comfort upon application to the skin. With a pH value of 6.8, the cream falls within the slightly acidic range, which is often preferred for skincare products as it is closer to the skin's natural pH, promoting compatibility and minimizing the risk of irritation. Moreover, the cream demonstrates excellent stability, scoring a rating of 100, suggesting it is robust and resistant to degradation over time, ensuring its efficacy and longevity. Overall, these characteristics suggest that the cream is well-suited for skin applications, offering hydration, comfort, and stability for daily use.

Wound Healing Activity

Wounds pose significant health challenges, impacting both morbidity and mortality. Effective wound healing is crucial for restoring the disrupted anatomical stability and functionality of the skin. Rapid healing requires swift wound contraction, a shorter epithelialization period, and sufficient tensile strength.^39,40

Herbal medicines are essential in both developed and developing countries for enhancing primary healthcare due to their potent biological and medicinal properties. H. scoparia, traditionally used in various regions of Ethiopia, has shown promise. ⁴¹ Acute dermal toxicity tests revealed no signs of toxicity over a 14-day period, indicating the test samples were non-irritating according to the dermal irritation scoring system. Therefore, H. scoparia extract can be safely used as a topical treatment for wounds.^42,43

Various creams were applied to the wounds in all groups, and then the wound dimensions were measured after every three days to observe the wounds macroscopically, and then the healing rate was calculated each time to observe the speed of the healing effect of the creams carried out in this study using H. scoparia extract and DOUCE Plus compared to the negative control groups on the wounds. Figures 2 and 3 presents the wound healing rates in albino rats over 4, 7, 10, and 13 days using different treatments:

On day 4, H. scoparia cream showed a wound healing rate of 49.20%, the highest among all treatments, indicating early efficacy. DOUCE PLUS cream had a healing rate of 30.98%, which was lower than the other creams but higher than the no cream treatment, while the no cream treatment resulted in a healing rate of 34.31%, the lowest on day 4.

By day 7, the H. scoparia cream continued to outperform other treatments with a healing rate of 58.76%. DOUCE PLUS cream showed a notable increase to 60.98%, while the no cream treatment had a rate of 49.88%.

On day 10, H. scoparia cream reached a significant healing rate of 92.09%, indicating its high effectiveness in promoting wound healing over time. DOUCE PLUS cream had a healing rate of 77.65%, while the base cream treatment showed a rate of 79.76%. The no cream treatment lagged with a rate of 59.76%.

By day 13, H. scoparia cream achieved the highest wound healing rate of 96.54%, demonstrating its superior efficacy in wound healing. DOUCE PLUS cream had a rate of 88.98%, indicating good effectiveness. The no cream treatment had the lowest rate of 66.54%.

Overall, these results suggest that H. scoparia cream is the most effective treatment for promoting wound healing in albino rats. The DOUCE PLUS cream also showed good efficacy, while the base cream and no cream treatments were less effective. The data highlights the potential of H. scoparia-based creams as potent wound healing agents.

Hematological Parameters

On day 13 of the study, the hematological parameters were analyzed across various treatment groups, shedding light on the effects of different creams on blood composition. The mean levels of red blood cells (RBCs) varied slightly among the groups (Figure 4), with the H. scoparia cream showing the highest mean level at 5.2 × 10^6/µL. Conversely, the DOUCE PLUS cream exhibited the lowest mean RBC level at 4.8 × 10^6/µL. Similarly, white blood cell (WBC) levels (Figure 4) showed marginal differences, with DOUCE PLUS cream displaying the highest mean levels, while the No cream group exhibited the lowest. Blood platelet (PLT) levels (Figure 4), an essential factor in clotting, also varied across groups, with H. scoparia cream demonstrating the highest mean level at 250 × 10^3/µL. Overall, these findings provide insights into how different topical treatments may influence hematological parameters.

Figure 4.

Mean of RBCs, WBCs, and PLT levels on day 13 of treatment groups: H. scoparia, DOUCE PLUS, and No cream; n = 5.

Inflammation Parameters

The inflammation parameters assessed on day 13 offer crucial insights into the inflammatory response within each treatment group. C-reactive protein (CRP) levels (Figure 5), indicative of systemic inflammation, exhibited slight variations across the groups, Hammada scoparia and DOUCE PLUS cream displaying marginally higher mean levels compared to others. Conversely, the No cream group showed the highest mean CRP level, suggesting a potentially heightened inflammatory state in the absence of topical treatment. Sedimentation rate (ESR), a non-specific marker of inflammation represented in Figure 5. These findings underscore the potential influence of topical creams on inflammation parameters, highlighting the importance of considering both hematological and inflammatory factors in assessing treatment efficacy and safety.

Figure 5.

Mean of C-RP, SR Levels on day 13 of treatment groups: H. scoparia, DOUCE PLUS, and No cream treatments; n = 5.

Histological Study

The study revealed notable alterations in the overall appearance of the skin, which were contingent upon various developmental stages and healing scenarios induced by the application of cream formulations. Specifically, the utilization of H. scoparia extract and DOUCE Plus creams, elicited distinct responses compared to the negative control groups. These treatments appeared to influence the skin's histological features, prompting changes indicative of potential therapeutic effects. The ensuing figures present detailed insights into the histological alterations observed across the different treatment groups, shedding light on the underlying mechanisms driving the observed changes in skin morphology and healing processes (Figure 6).

Figure 6.

Microscopic observation of a histological section of skin wound in different experimental groups, No Wound + No cream treatment group (A), Wound + H. scoparia cream treatment group (B), Wound + DOUCE PLUS^® cream treatment group (C), and Wound + No cream treatment group (D), magnification ×40. Legends:1- Epidermis, 2- Dermis, 3- Adipose tissue, 4- Stratum corneum, 5- Hair follicles, 6- Sweat gland.

For No Cream Treatment Group, the untreated wound exhibited a marked inflammatory response with extensive inflammation, dense infiltration of inflammatory cells, less organized collagen fibers, and signs of necrosis in the epidermal layer. Healing was delayed compared to treated samples. The histological section of the untreated wound showed more disruption and irregularity in the epidermis, with areas of necrosis and sloughing. The dermis has extensive inflammation, with dense inflammatory cell infiltration, disorganized collagen fibers, and edema. The subcutaneous adipose tissue appears less affected. Overall, healing is delayed in the untreated wound, with slower re-epithelialization, significant tissue disruption, and less organized collagen and fibroblast activity compared to treated samples. There are also fewer new blood vessels, suggesting reduced support for tissue repair.

For Hammada scoparia Cream Treatment Group, the histological section epidermis showed evidence of re-epithelialization with a more organized layer. The dermis had moderate fibroblast activity and collagen deposition, indicating ongoing tissue repair. Inflammatory response was moderate.^44,45 This could be attributed to the plant extract's capacity to promote collagen synthesis, stimulate cell proliferation, and exhibit antimicrobial activities through its bioactive constituents. ⁴⁶ This may be attributed to the astringent, antioxidant, and antibacterial properties of phytoconstituents such as phenols and tannins in the water and ethyl acetate fractions, which promote the regeneration of new tissue. ⁴⁷ Phenolic compounds, such as tannins, substituted cinnamic acids, phenolic acids, and phenylpropanoids, have demonstrated both antimicrobial and antioxidant properties. ⁴⁸ Tannins, in particular, are known to promote wound healing. ⁴⁹ These polyphenolic compounds are rich in hydroxyl, carboxyl, and other hydrophilic groups, and are classified as macromolecules due to their large molecular structure. Additionally, compounds like caffeic acid, chlorogenic acid, and ferulic acid are documented to exhibit wound healing activities.^50,51

For DOUCE PLUS^® Cream Treatment Group, the epidermis appeared partially intact with some irregularities. The histological section dermis showed increased fibroblast activity and densely packed collagen fibers, indicating active repair. Numerous blood vessels were present, suggesting angiogenesis. Signs of inflammation were also observed. ⁵²

Overall, histological observations suggest that H. scoparia cream and DOUCE PLUS^® cream facilitated wound healing by promoting re-epithelialization, fibroblast activity, collagen deposition, and angiogenesis, with H. scoparia cream showing a more organized and effective healing process. The study's limitations include the lack of quantification of phenolic compounds in Hammada scoparia extract, absence of accelerated stability testing, and histological analysis at a single time point. Additionally, the focus on wound size as the primary healing parameter overlooked other aspects like wound depth and collagen deposition. These limitations highlight areas for future research.

Conclusion

The present study demonstrated that Hammada scoparia aerial parts exhibit rapid wound healing activity compared to the negative control group. The presence of bioactive metabolites likely contributes to this wound healing effect. Therefore, the results indicate that the aerial parts of Hammada scoparia possess wound healing properties, supporting its traditional use for treating wounds as documented in traditional medicine literature.