EVALUATION OF ANTI-ACNE POTENTIAL OF IMPATIENS WALLERIANA EXTRACT

EVALUATION OF ANTI-ACNE POTENTIAL OF IMPATIENS WALLERIANA EXTRACT

G. Kulkarni *, K. Tiwari, R. Yadav and K. Vaibhavi

Department of Pharmacology, Rani Chennamma College of Pharmacy, 7/C, B. K. Kangrali, Industrial area, Vaibhav Nagar, Bauxite Road, Belagavi, Karnataka, India.

ABSTRACT: Objective: Knowledge about medicinal plants can contribute to modern therapeutics. Impatience walleriana [Family: Balsaminaceae] an ornamental, nutritive herb has relevance in African and Asian traditional medicines. Traditionally I. walleriana is used for anti-inflammatory, anti-dermatitis effects and is known to relieve skin irritation. It is applied on skin to treat bacterial and fungal infection. Acne is a common skin disorder mainly seen in adolescents. Treatment for acne revolves around anti-bacterial, anti-inflammatory and antioxidant effects of drugs. Anti-bacterial and antioxidant activities of I. walleriana have been reported previously reported. In the present study anti-acne potential of I. walleriana extract is evaluated against Propionibacterium acne and an attempt has been made to co-relate findings with traditional claims. Methods: Hydroalcoholic extract of aerial parts of I. walleriana is evaluated for effectiveness against P. acne by measuring zone of inhibition using well diffusion method and by finding MIC. Tetracycline is used as standard. Results: Results of Zone of Inhibition are promising with an average of 11mm, 08mm and 4mm in the duplicate readings obtained. MIC results with 1.6µl/ml concentration reinforces the effectiveness of I. walleriana extract against P. acne organism. Conclusion: Our hypothesis is met with positive results. I. walleriana extract can be a good source for supplementing existing anti-acne medicines.

Keywords: Antibacterial, Propionibacterium acne, Traditional medicine, Acne

INTRODUCTION: Acne is a common skin affliction, encountered by adolescents and many adults continue to struggle even after their teenage years. Though acne is not a lethal or debilitating disorder, it is painful and disfiguring ¹. It affects confidence in individuals,, reducing self-esteem and causing emotional distress ². Acne is a common skin disorder, almost affects everybody during lifetime ³. Acne is characterized by inflammatory lesions or comedones in the face, back and trunk regions ⁴. Acne vulgaris is multifactorial in origin ⁵.

Genetics plays an important role, and acne develops as a result of follicular epithelium hyper-keratinization, increased sebum production, proliferation of P. acne like bacteria and inflammation. P. acne, Staphylococcus epidermidis and S. aureus are involved in the bacterial colonization of the follicle. Among the three organisms P. acne plays an important role in pathogenesis and progression of acne ⁶.

The other two organisms mainly contribute by forming a biofilm which provides anaerobic conditions for P. acne to thrive ⁶. Reactive oxygen species (ROS) are subsequently generated from hypercolonization of P. ance and from UV exposure ⁷. Sebum also contains ROS and these free radicals also contribute to irritation of acne ⁸. Current therapies for acne by topical and systemic routes employ use of antibiotics, comedolytics, anti-inflammatory drugs, zinc in addition to laser treatment. The currently used drugs and approaches have limitations in the form of toxicity, side effects and high costs ⁸. Among the prevailing treatments not a single is full-proof and satisfactory ³. P. acne is also showing resistance to the currently used antibiotics ¹. Owing to these problems novel new therapies are in demand.

Traditionally medicinal plants have been used for managing a number of dermatological conditions ¹. Valuable leads can be obtained from herbal resources ¹. Folklore use of medicinal plants can serve as a guide to identify bioactive compounds and is useful compared to random search.¹ Traditional herbs are also unexplored sources for developing new drugs ².

Herbs can play an important role in finding new lead compounds for acne. Natural remedies are blooming as they target multiple factors related to acne ⁹. Recent trends show use of plant-based products as adjunctive and alternatives to current therapies ¹⁰. Herbal approaches are also relatively free of side effects and are economical ¹¹. It is estimated that nearly 20% of all visits to dermatologists are related to the treatment of acne ¹².

Genus Impatiens is mainly known for antioxidant and antimicrobial activity. It is also recognized for antiallergic, antipruritic, antidermatitic, antihistaminic and COX-2 inhibitory activities ¹³. I. walleriana is less researched compared to other Impatiens species. Impatiens treats bruises. I. walleriana leaves are used to treat chicken pox in Indonesia ¹³. In Costa Rica flowers are used to heal wounds by rubbing onto the wound ¹³.

I. walleriana Hook.f is an ornamental plant belonging to family Balsaminaceae. It is both nutritive and medicinal. It finds use in Brazilian folk medicine ¹⁴. Antioxidant, antibacterial and antifungal activities of the plant are reported but it has not been screened for effectiveness against P. acne hence the present study.

MATERIALS AND METHODS ¹⁵:

Collection of Plant and Extraction: The fresh leaves, stem and pink flowers of I. walleriana were collected from a local garden in Belagavi during the month of July 2024. Authentication for the plant was carried out at Regional Medical Research Centre, Belagavi and specimen was deposited in the said research centre herbarium (Accession number RMRC – 1341). The fresh plant parts were first washed using tap water and soiled portions were cleaned to exclude root portion followed by drying in shade at room temperature. The dried plant material was size reduced to coarse powder and stored in an air tight bottle.

Extraction was done using 50:50 Distilled water and 95% alcohol as solvent by maceration for 72 hours. After extraction the solvent was evaporated, and residue was collected then % yield was calculated using the formula:

% Yield = W₁ / W₂ × 100

Where, W₁ = weight of the extract after extraction, W₂ = weight of plant powder.

Well Diffusion Assay Procedure: The well diffusion test was performed to evaluate the antimicrobial activity of the compounds. Initially, colonies from the agar plate were transferred to plates using a swab. The inoculum was prepared by adjusting the turbidity to equal that of a 0.5 McFarland turbidity standard visually.

Within 15 minutes, a sterile cotton swab was dipped into the inoculum, excess removed, and the agar plate was swabbed three times, rotating 60° between streaks. The inoculated plate stood for 3-15 minutes before making wells.

Stock solutions were prepared by dissolving 10mg of compound in 1ml of Dimethyl sulfoxide. Four wells (5mm diameter) were created on each plate using a heated hollow tube. Using a micropipette, 200μl, 100μl and 50μl of the stock solution were added to each well and tetracycline was added to the fourth well.

Plates were incubated within 15 minutes, inverted, stacked (no more than five high), and incubated for 18-24 hours at 37°C. After incubation, plates with confluent or nearly confluent lawns were read. The diameter of the inhibition zone was measured to the nearest whole millimeter using a measuring device.

MIC Test Procedure: The Minimum Inhibitory Concentration (MIC) test was performed using the broth dilution method. For I. walleriana extract, nine serial dilutions were prepared in Brain Heart Infusion (BHI) broth.

Initially, 20μL of the drug was added to 380μL of BHI broth in the first tube. Then, 200μL of BHI broth was added to each of the remaining nine tubes. Serial dilutions (10-1 to 10-9) were prepared by transferring 200μL from the previous tube to the next, starting from the initial tube.

5µl of the test organism containing stock culture was inoculated into 2ml of BHI broth and incubated. Subsequently, 200μl of this culture suspension was added to each serially diluted tube. The tubes were incubated for 24 hours and observed for turbidity to determine the MIC.

RESULTS AND DISCUSSION:

Extract Yield: I. walleriana extract exhibited consistency that was resinous and appearance was brown with earthy odour. The yield of the extract was found to be 15.36 g%.  Zone of inhibition has been carried out twice and duplicate readings are depicted as IWA I and IWA II:

FIG. 1A: IWA I FRONT VIEW                     

FIG. 1B: IWA I BACK VIEW

FIG. 2A: IWA II FRONT VIEW                        

FIG. 2B: IWA II BACK VIEW

TABLE 1: RESULTS OF WELL DIFFUSION ASSAY

R= Resistant.

FIG. 3: WELL DIFFUSION METHOD

MIC Results:

FIG. 4: MIC TEST RESULTS

TABLE 2: RESULTS OF MIC

S=Sensitive; R=Resistant; B+C = Broth + Control; B+O = Broth + Organism, P= Present.

Medicinal plants with their chemical constituents offer high therapeutic benefits. Several plants have been researched for treatment of diseases. To resolve concerns like antibiotic resistance herbal Plants with improved safety and efficacy are handy. If proven effective the plant can become a supplement to the current antibacterial therapy ¹¹.

P. acne is slow growing, aero tolerant–anaerobic rod shaped gram positive bacterium associated with acne. It resides deep inside in follicles and pores where these bacteria use sebum and metabolic by-products for their survival. Hyperactive sebaceous gland, obstruction of the follicle like factors can significantly increase growth of P. acne. In addition to P. acne, M. furfur, an yeast and S. epidermidis are also present in acne lesions. S. epidermidis is gram positive, aerobic and found in affected acne pores in addition to P. acne ¹⁰. S. aureus is also involved in triggering inflammatory acne flare-ups ⁶. Reactive oxygen species are generated from hyper-colonization of P. acne and from exposure to UV-rays. Herbals inhibiting these organisms and having antioxidant activity are of great interest in treating acne ⁷. Herbals with healing multiple factors related to acne are in demand ³. The objective of the present study was to investigate effectiveness of I. walleriana extract against acne causing P. acne organism and confirm the folklore use of I. walleriana for various skin related afflictions. Such studies backed by folk use and traditional medicinal claims are better than random screening of plant extract to find new medicinal alternatives. Our hypothesis that I. walleriana extract being antioxidant, antibacterial, anti-inflammatory might be effective against P. acne is well substantiated in our present investigation wherein antibacterial effect is confirmed through MIC as well as Zone of inhibition study. In our previous findings we learnt about the effectiveness of I. walleriana extract against S. epidermidis and S. aureus the other organisms involved in acne development ^{11, 16}. I. walleriana extract is effective against all the three bacteria causing acne namely P. acne, S. epidermidis & S. aureus making it an interesting contender for developing an alternative for acne. This is also the first study to document I. walleriana extract effect against P. acne.

Results of Zone of Inhibition are promising with an average of 11mm, 8mm and 4mm in the duplicate readings obtained. Effectiveness of the standard tetracycline is much higher than I. walleriana extract nevertheless I. walleriana extract in principle can constitute adjunctive to currently used antimicrobials for the treatment of acne. A dose dependent increase in antibacterial effect has been observed in the study. MIC results with 1.6µl/ml concentrations show the effectiveness of I. walleriana extract against P. acne organism. According to Gibbson and researchers isolated phytochemicals should have MIC < 1mg/ml ⁷. In this study the constituents have not been isolated as such but extract has been studied. MIC results of I. walleriana reinforce antibacterial effectiveness. The plant is reported to have Flavonoids, Alkaloids, Carbohydrates, Tannins, Anthocyanins, Caffeic acid, minerals such as calcium, phosphorous, potassium, sodium, chloride, magnesium, iron, zinc, iodine, chromium, copper, fluoride, molybdenum, manganese and selenium etc ^{11, 13}. In the present study the observed effect may be due to these constituents showing effect including synergistic effect which cannot be ruled out. Presence of zinc like constituents in the extraxt may be contributing to antiacne effect by anti-inflammatory action. Generally polar organic solvents are used to obtain extract high in Phenolic compounds supporting antioxidant effect, hence in the present study hydroalcoholic (50:50) solvent has been employed for extraction. Anti-acne drugs have side effects like skin drying and also have other toxicities. Acne infection is also associated with irritation. I. walleriana as per traditional claims offers advantage in terms of resolving skin irritation. Further studies can be taken up to validate the claim. Additionally effectiveness of I. walleriana extract against the yeast implicated in acne M. furfur can be taken up. As I. walleriana extract effectiveness against P. acne is learnt through this study, clinically resistant strains of P. acne can be subjected to testing specifically. I. walleriana extract being effective against S. epidermidis, S. aureus and P. acne may benefit inflammatory acne in papules, pustules and reduce bacterial infection which leads to abscess development. I. walleriana extract can serve as an alternative to supplement existing antibacterials for acne treatment following evaluation of all parameters of anti-acne efficacy.

CONCULSION: Our hypothesis is met with positive results for anti-acne activity of hydro-alchoholic extract of I. walleriana. I. walleriana extract can be a good source to supplement existing anti-acne medicines. Further studies are required to evaluate other parameters of anti-acne efficacy and find the exact mechanism of action.

ACKNOWLEDGMENTS: The authors extend their gratitude to college authorities for providing facilities. We also acknowledge Dr Harsha Hegde, Scientist, RMRC for helping with authentication of the plant and Dr Ulka Idate, Department of Microbiology, CRL, Maratha Mandal’s Institute of Dental Sciences and Research, Belagavi for helping with antimicrobial screening. We also acknowledge Miss Sushmita Adibatti and Mr Veeresh Nagashetti for assistance in the laboratory.

CONFLICT OF INTEREST: Nil

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    How to cite this article:

    Kulkarni G, Tiwari K, Yadav R and Vaibhavi K: Evaluation of anti-acne potential of Impatiens walleriana extract. Int J Pharmacognosy 2025; 12(6): 503-08. doi link: http://dx.doi.org/10.13040/IJPSR.0975-8232.IJP.12(6).503-08.

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