Development of antimicrobial resistance in bacteria is a major concern. The study of quorum sensing (QS), the bacterial communication system that coordinates important events in an infection, opens a new avenue for treating infections. QS helps in biofilm formation which acts as a physical barrier and alters the microenvironment which traps the molecules or slows down the antibiotic diffusion making them less effective. Since the standard drugs are not very effective against bacteria anymore, demand for the discovery of other alternatives arises. Phytochemicals extracted from commonly used plants like Coleus amboinicus, known to have antimicrobial properties, can be used to develop novel drugs against quorum sensing and hence suppress bacterial infection. Here, we have identified a set of phytochemicals from C. amboinicus whose drug-like properties were predicted using SwissADME and their interactions with the targets involved in QS in Pseudomonas aeruginosa, Vibrio harveyi and Chromobacterium violaceum were studied by performing molecular docking using PyRx software. Beta-amyrin, chrysoeriol and apigenin were found to be the top scored molecules with docking scores of -9.3, -11.3, and - 11.1kcal/mol respectively, and were considered for molecular dynamics simulation using GROMACS software. This was followed by in vitro tests such as Agar well diffusion, Scanning electron microscopy, Pyocyanin assay and Crystal violet assay. Erythromycin was used as the standard drug and as a positive control to validate the results. Based on both in vitro and in silico studies, the phytochemicals from C. amboinicus have the potential to be inhibitors of quorum sensing targets. These findings are preliminary and should be interpreted as hypotheses to be tested experimentally rather than as established mechanisms.

Plectranthus amboinicus leaves are rich in bioactive phytochemicals with documented antioxidant and anti-inflammatory activities. The present study was conducted to discuss the protective potentials of free ethanolic extracts of P. amboinicus leaves (PAEE) and its chitosan (CS)-based nanoformulation against lambda-cyhalothrin (LCT)-induced oxidative stress, inflammation, and DNA damage in male mice. Animals were equally distributed into five experimental groups as follows: control, LCT-treated (1 mg/kg b.wt), PAEE (200 mg/kg b.wt) + LCT, CS-loaded PAEE nanoformulation (200 mg/kg body weight) + LCT, and CS nanoparticle control. All treatments were administered orally for 28 days. At the end of the experiment, blood samples were collected and serum samples were analyzed for oxidative stress markers (SOD, CAT, GPx), pro-inflammatory cytokines (TNF-α, IL-6, IL-33, IL-17), the autoimmune antinuclear antibody marker (ANA) and hematological parameters. Moreover, DNA damage was assessed by comet assay. Spleen tissues were harvested for histopathological examination. Results of the present study revealed that LCT exposure resulted in significant oxidative stress, evidenced by reduced antioxidant enzyme activities (SOD, CAT, GPx), elevated pro-inflammatory cytokines (TNF-α, IL-6, IL-33, IL-17) and ANA, altered hematological parameters, increased DNA damage, and histopathological alterations in spleen tissue. Pretreatment with PAEE significantly attenuated these effects, while the nanoformulation demonstrated superior protective efficacy compared to the free extract. The enhanced efficacy of CS/PAEE may be attributed to improved stability, bioavailability, and sustained release of PAEE bioactive compounds resulting in prolonged antioxidant and anti-inflammatory effects. These findings demonstrate that P. amboinicus-based nanoformulations may represent a promising, eco-friendly approach for mitigating pesticide-induced toxicological effects associated with environmental exposure.

The pursuit of eco-friendly nanomaterials with both antibacterial and anticancer properties is gaining momentum in biomedical research. This study reports the green synthesis of Silver and Zinc oxide (Ag/ZnO) nanocomposite using Coleus amboinicus leaf extract as a natural reducing and stabilizing agent. Characterization via UV–Vis spectroscopy revealed absorption peaks at approximately 280 nm and 370 nm, indicative of Ag and ZnO nanoparticles, respectively. FTIR analysis identified phytochemical-derived functional groups responsible for particle stabilization, while SEM imaging showed agglomerated nanoparticles with diameters ranging from 100 to 150 nm. XRD analysis confirmed the crystalline nature of the nanocomposites. The biosynthesized Ag/ZnO nanocomposite exhibited potent antibacterial activity against multidrug-resistant strains including Enterococcus faecalis , Pseudomonas aeruginosa , MRSA, and Klebsiella pneumoniae , with significant zones of inhibition observed at a concentration of 80 μg/mL. Furthermore, the nanocomposite displayed promising anticancer activity against A549 lung cancer cells, achieving an IC₅₀ of 60 μg/mL after 24 h, as determined by MTT assay. Morphological features of apoptosis, such as membrane blebbing and nuclear condensation, were observed, while AO/EtBr, DCFDA, and Rhodamine 123 staining confirmed elevated reactive oxygen species (ROS) generation and disruption of mitochondrial membrane potential. These findings suggest a ROS mediated mechanism underlying the dual antibacterial and anticancer activities of the Ag/ZnO nanocomposite. This green synthesis approach provides a sustainable strategy for developing multifunctional nanomaterials, although further in vivo studies are required to confirm efficacy and elucidate the molecular mechanisms involved for future therapeutic applications. • Green synthesis of Ag/ZnO nanocomposite using Camboinicus leaf extract • Potent activity against MDR bacteria at 80 μg/mL concentration • Anticancer efficacy against A549 lung cancer cells • ROS-mediated mechanism confirmed via multiple fluorescence staining assays.

Coleus amboinicus (Lour.), commonly called Indian borage, Mexican mint, and Oregano thyme, is a member of the Lamiaceae family, native to Southeast Asia and Africa. This herb is widely known for its rich nutritional composition, including essential vitamins, minerals, and bioactive compounds such as thymol, carvacrol, and rosmarinic acid. The therapeutic potential of C. amboinicus lies in its antioxidant, anti-inflammatory, antimicrobial, and prebiotic properties, which aid in managing chronic diseases such as cardiovascular issues, diabetes mellitus, and respiratory conditions. Recent studies have demonstrated its potential in food preservation, offering a natural alternative to synthetic preservatives due to its antimicrobial properties. Despite its promising applications, challenges such as variability in bioactive content, a lack of robust clinical trials, and regulatory approval remain. This review examines the nutritional composition, therapeutic benefits, and potential as a functional food of C. amboinicus. It suggests future research directions to overcome existing barriers and enhance integration into modern dietary systems.

Plant secondary metabolites provide a unique basis for medications, flavorings, and industrial biochemicals. These metabolites frequently accumulate in plants exposed to environmental stressors (e.g., drought stress), as a mechanism for adaptation and resistance. Plectranthus amboinicus (Lour.) Spreng is a semi-succulent Lamiaceae species valued for its curative properties and essential oil, the concentration of which is heavily impacted by irrigation levels. This investigation reveals a novel study into the physiological and molecular mechanisms of P. amboinicus by correlating field capacity (FC) levels with metabolite accumulation and specific gene expression. We examined the impact of six irrigation levels (0, 20, 40, 60, 80 and 100%; denoted as FC-0 to FC-100) on growth, essential oil (EO) yield, and carvacrol biosynthesis during the 2022 and 2023 seasons. Data revealed that plants subjected to the FC-0 exhibited the lowest growth parameters. However, these plants also presented a higher essential oil percentage and carvacrol concentrations, in addition to an increase in polyphenols and antioxidant enzymes, with greater oxidative substances accumulation (H2O2 and MDA). Moreover, these plants presented higher gene expression of CYP71D178 and CYP71D180 genes. While increasing water level caused an increase in growth parameters, with a decline in the antioxidant activity. Increasing the water level to FC-100 increased the essential oil by approximately four fold but resulted in a decline in carvacrol content as compared to FC-0. Generally, water deficiency reduces crop yield, whereas irrigation increases plant productivity. However, in medicinal and aromatic plants, the relationship is distinct; in Plectranthus amboinicus, water deficiency showed a strong positive correlation with the secondary metabolite’s generation. In conclusion, the current report provides for the first time a molecular framework for P. amboinicus by depicting the expression patterns of CYP71D178 and CYP71D180. We revealed that these genes elaborate as the primary molecular shifts regulating carvacrol biosynthesis influenced by specific irrigation levels.

Due to increasing insecticide resistance and environment concerns associated with synthetic chemicals, this study evaluates the larvicidal potential of essential oils (EO) extracted from seven plant species using hydro-distillation against Aedes aegypti larvae. The chemical profiles of the volatile oils were characterized through gas chromatography-mass spectrometry (GC-MS). The major constituents identified were thymol (74.93%) in Plectranthus amboinicus leaves, terpinen-4-ol (27.13%) in Myristica fragrans nuts, citral (27.55%) in Citrus limon leaves, caryophyllene oxide (36.11%) in Hyptis suaveolens leaves, and asarone (85.51%) in Acorus calamus rhizomes. Larvicidal bioassays revealed LC50 values below 50ppm (23.40 to 47.34ppm), with the order of efficacy being A. calamus > M. fragrans > T. ammi > I. verum > C. limon > H. suaveolens > P. amboinicus. Twenty-one binary and thirty-five ternary combinations were tested to determine synergistic, additive, and antagonistic interactions using Wadley's, Ting-Chao Chou, Co-Toxicity Factor, and Chi-square comparison methods. Binary combinations, such as P. amboinicus and M. fragrans and T. ammi and M. fragrans exhibited better synergism, this combination analysed by GC-MS, P. amboinicus and M. fragrans contains, carvacrol (36.23%), thymol (39.04%), and caryophyllene oxide (3.84%). Similarly, the combination of T. ammi and M. fragrans showed thymol (61.96%) and caryophyllene oxide (7.19%). Based on the EO yield and activity the T. ammi and M. fragrans binary combination formulated as soft-gel capsules, this formulation showed 100% larval mortality. EO mixtures work very well, showing they could be good, eco-friendly options to replace chemical insecticides for controlling mosquito larvae in the future.

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The objective of this study was to evaluate essential oils (EOs) as an alternative control method for Rhipicephalus microplus and Amblyomma mixtum larvae. The EOs used were obtained by steam distillation from the leaves of cinnamon (Cinnamomum verum), mexican mint (Plectranthus amboinicus), lemongrass (Cymbopogon citratus), peppercorns (Pimenta dioica), and peruvian pepper tree (Schinus molle). To evaluate the acaricidal effect of EOs against the larvae of two tick species, a larval immersion test (LIT) was performed using six concentrations (10 mg/mL, 5 mg/mL, 2.5 mg/mL, 1.87 mg/mL, 1.25 mg/mL, and 0.6 mg/mL), in addition to a negative control group (water + ethanol) and three positive controls (organophosphate, formamidine, and pyrethroid). LIT results were obtained 48 h after exposure. Data were processed using Probit procedure to determine the lethal concentrations at 50% (LC50), 95% (LC95), and 99% (LC99). For R. microplus, 99% mortality was obtained at concentrations as low as 1.4 mg/mL for S. molle, while the highest LC99 was recorded with P. dioica at 23 mg/mL. In the case of A. mixtum, higher concentrations were required to achieve a high mortality rate. EO of P. amboinicus had the lowest acaricidal effect, requiring 26.2 mg/mL to achieve an LC99, while S. molle required a concentration of 6.9 mg/mL to achieve an LC99.

Systematic investigation of aflatoxigenic Aspergillus flavus inhibition: Integrating essential oils-based formulation with mathematical modeling and transcriptomic analysis.

The escalating antimicrobial resistance (AMR) crisis necessitates the development of innovative anti-infectives with novel mechanisms of action. Nevertheless, research on natural products remains constrained by low-throughput screening and limited mechanistic insights. Artificial intelligence (AI) is catalyzing a pivotal paradigm shift-from the mere isolation of active compounds to precisely deciphering their modes of action. This review highlights AI's transformative role in bridging ethnopharmacological knowledge and modern pharmacology to decode the mechanisms of plant-derived anti-infectives. Case studies on berberine, baicalein, danshensu derivatives, and rosmarinic acid derivatives from Coleus amboinicus illustrate AI's capacity to map traditional therapeutic concepts to specific pathways (e.g., biofilm inhibition, inflammasome modulation) and to predict precise binding interactions and pharmacophores with high precision. Leveraging statistical correlations between ethnobotanical usage patterns and chemical similarity, we propose a "Knowledge-Data-Mechanism" three-layer framework centered on deep mechanistic insight. Integrating Chinese initiatives, such as the CNDR (China's National Drug Repository) database and the TCM-AI platform, with global traditional medicine wisdom, this strategy provides an actionable roadmap for modernizing anti-infective discovery. Validated applications of this paradigm have demonstrated order-of-magnitude acceleration in mechanistic characterization, rapidly yielding structurally novel agents with well-defined, target-specific actions-a critical advancement in addressing the urgent global threat of antimicrobial resistance.

Objective: Green synthesis of metal nanoparticles provides an environmentally friendly approach in comparison with the chemical method. Iron oxide nanoparticles (FeONPs) have potential biomedical applications such as antimicrobial, antioxidant, and anti-inflammatory activities. The goal of the current study was to synthesize FeONPs using the leaf extract of Plectranthus amboinicus and to assess the biological activities. Methods: FeONPs were prepared in a green reduction and stabilization method using aqueous extracts of P. amboinicus leaves. The formation of FeONPs was initially qualitatively identified through colorimetry and additionally identified using ultraviolet (UV)-visible spectroscopy, X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). Antimicrobial testing was conducted against Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus mutans, Aggregatibacter actinomycetemcomitans, and Candida albicans using minimum inhibitory concentrations (MIC) and time-kill methods. The antioxidant property of the synthesized FeONPs was evaluated using the 2,2-diphenyl-1-picrylhydrazyl radical scavenging assay. The anti-inflammatory property was analyzed by protein denaturation inhibition assays utilizing bovine serum albumin and egg albumin models. The cytotoxicity as well as the toxic properties of FeONPs were analyzed by brine shrimp (Artemia salina) lethality bioassays. Results: The emergence of a clear color transition from dark brown to light brownish-orange signified nanoparticle development, accompanied by a distinctive UV–Visible Spectroscopy absorption wavelength at 395 nm. XRD pattern verification demonstrated the nanocrystalline and phase-pure quality of FeONPs, measuring 10–12 nm. FTIR pattern matching further demonstrated surface modification by OH, aromatic, and phenolic moieties. The MIC range was 25–100 μg/mL, demonstrating broad-spectrum antibacterial activity, considerable antioxidant activity, and moderate anti-inflammatory activity. Cytotoxicity studies also exhibited moderate cell toxicity with LC50 = 8 μg/mL. Conclusion: Green-synthesized FeONPs using P. amboinicus demonstrated stability, multifunctional bioactivity, and promising antimicrobial, antioxidant, and anti-inflammatory properties, highlighting their potential for further in vivo biomedical applications.

Intensive industrial activities have led to increased chromium concentrations in soil and water, posing significant environmental and health risks due to its cytotoxic and carcinogenic nature. Present study is aimed to elucidate the sequel of chromium toxicity on structural and functional properties of Coleus amboinicus Lour. The experimental set up consisted of cultivation of rooted propagules of C. amboinicus in nutrient solution artificially contaminated with 150 µM potassium dichromate in which the plants exhibit visible growth retardation and maintaining their survival. Structural alterations of plants treated with Cr studied in terms of anatomy as well as tissue ultra structure using Scanning Electron Microscopy. Presence of Cr was observed in the form of stained masses in the sections of root, stem and leaf, indicating the translocation of Cr to the plant body. Qualitative and quantitative detection of elemental composition of both control and experimental were made by energy dispersive X-ray analysis. Cr influence the macro and micro-elemental distribution in plant tissues. Plants exhibited structural modifications like increase in the trichomes of leaves and stem of Cr treated C. amboinicus. Presence of cell structural distortions and Cr deposit inclusions in the inner parenchyma cells were distinct. Cr stress induced the reduction in pigment content and metabolites like proteins, proline, phenol and malondialdehyde marked a significant increase. The low BCF and TF values, combined with its coping mechanisms for metal stress, indicate that C. amboinicus lacks the potential for chromium hyperaccumulation.

This study investigated the development of a mouthwash formulated with Plectranthus amboinicus leaf extract (PaEE) and Lippia sidoides leaf essential oil (LsOE), exploring their antibacterial potential. Antimicrobial activity was evaluated in vitro using the broth microdilution method to determine the minimum inhibitory concentration (MIC) against Streptococcus mutans and Staphylococcus aureus. Chemical analysis of LsOE by GC/MS identified active compounds such as thymol and carvacrol, while PaEE demonstrated the presence of flavonoids, anthocyanins, mucilages, tannins, and phenolic compounds. Both extracts exhibited antibacterial activity, with LsOE and the natural mouthwash showing intermediate efficacy (MIC 128µg/mL), while PaEE alone was less effective (MIC 512µg/mL). The formulated mouthwash (F1) significantly reduced bacterial growth, and the combined use of LsOE and PaEE indicated a synergistic effect. These findings support the potential of plant-based formulations for dental biofilm control and encourage further research to expand their applicability in public oral health.

Retraction notice to "Antibacterial, antifungal, antidiabetic, and antioxidant activities potential of Coleus aromaticus synthesized titanium dioxide nanoparticles" [Environ. Res. 216 (2023) 114714

Tilapia Lake virus (TiLV) is an emerging pathogen causing up to 90% mortality in tilapia (Oreochromis niloticus) farming. Enhancing tilapia health by modulating the non-specific immune system with herbal immunostimulants offers a promising strategy to mitigate TiLV outbreaks. This study evaluated the prophylactic efficacy of Gooseberry (Phyllanthus acidus) and Oregano (Coleus amboinicus) extracts against TiLV. Crude ethanolic extracts were prepared by Soxhlet extraction and screened for preliminary phytochemicals. The crude extracts were incorporated into the commercial pellet feed at varying concentrations (100, 300 and 500mg/kg) to prepare the experimental diets (PA (P. acidus), CA (C. amboinicus) and PC (P. acidus + C. amboinicus)) and fed to healthy tilapia for four weeks. They were then challenged with TiLV (intra-peritoneal) to assess the prophylactic effects. The combination diet (PC) at 300mg/kg showed the highest relative percentage survival (RPS) of 77.78%, followed by CA at 500mg/kg. Further, fish fed a diet containing PC at 300mg/kg showed significant (P < 0.05) improvements in haematological parameters, immune parameters, and immune gene expression (TLR-7 and IFN-β) at 24, 48, and 72h post-infection (hpi). Notably, a significant increase in blood performance, antioxidant defence and expression of IFN-β and TLR-7 in the spleen was observed. This study emphasises the potential of herbal-based immunostimulants for developing sustainable strategies to improve fish health and resilience against TiLV.

A novel macrophage-modulating cream containing proprietary extracts from Plectranthus amboinicus and Centella asiatica has been approved for the treatment of diabetic ulcers in some countries. It has been reported to suppress M1 macrophages and facilitate the transition from M1 to M2 macrophages, reducing inflammation and leading to faster wound healing. Based on its mode of action, it has also been used in non-diabetic wounds. This report describes the treatment of a patient with essential thrombocythaemia (ET) presenting initially as a toe ulcer, and which was successfully managed with the cream before the use of systemic therapy for the ET.

Unlocking the phytochemical and pharmacological potential of Ocimum sanctum L. and Plectranthus amboinicus (Lour.) Spreng. toward developing novel evidence-based herbal products in Sri Lanka: A comprehensive review

Background and ObjectiveStaphylococcus aureus poses a significant public health threat globally, where both community and hospital-acquired infections are prevalent. The escalating antimicrobial resistance highlights the urgent need for alternative therapies. Hence, traditional medicine using plant extracts offers a potential avenue for novel antibacterial agents. This systematic review aimed to evaluate the existing literature on the antibacterial properties of Philippine plants against S. aureus to provide focus on drug development of a plant-derived antibacterial for this pathogen.MethodsFollowing PRISMA guidelines, a comprehensive search was conducted in PubMed/Medline, SCOPUS, and Herdin databases. Inclusion criteria encompassed in vitro studies evaluating the antibacterial activity of crude plant extracts sourced from Philippine plants against S. aureus. Data extraction and quality assessment were performed independently by two reviewers, with discrepancies resolved by the third and fourth reviewers.ResultsOf the 413 initial studies identified, nine met the eligibility criteria. The highest zone of inhibition was demonstrated by Lippia micromera leaf essential oil at 26.3±1.5 mm, while moderate antibacterial activity was shown by essential oils from Alpinia elegans, Piper quinqueangulatum, and Alpinia cumingii at MIC values of 512 μg/mL, 512 μg/mL, and 1,024 μg/mL, respectively. Other Philippine plants showed a wide range of activity, with MIC values between 50 μg/mL and 25 mg/mL, MBC values from 78 to 5000 μg/mL, and ZOI ranging from 5 to 38 mm. However, the overall quality of evidence in these other studies are compromised by bias and incomplete reporting.ConclusionLeaf essential oils from Alpinia elegans, Piper quinqueangulatum, and Alpinia cumingii demonstrated moderate antibacterial activity against S. aureus. Additionally, the essential oils of Lippia micromera, Plectranthus amboinicus Lour. Spreng, and Cymbopogon citratus exhibited antibacterial activity against both S. aureus and Methicillin-resistant S. aureus (MRSA) in disk diffusion assays, these antibacterial activities may be attributed to their high concentrations of terpenes, terpenoids, and phenolic compounds. Majority of the studies gathered had high risk of bias according to the quality assessment criteria tool used in the study. Thus, this systematic review also emphasizes the need for improved methodological rigor on reporting in vitro antibacterial studies.

Introduction Lamiaceae is the name of a family of aromatic plants, which are traditionally used as herbal medicine. There are not many research articles found in the literature regarding the biological study of nanoforms of the Lamiaceae family. However in recent years, there were few studies exhibiting the biological properties of plants from the same family. The stem and leaves of three plants from Lamiaceae family of Plectranthus genus, namely Coleus amboinicus Lour , Coleus rotundifolius Lour, and Coleus zeylanicus Lour were subjected to extraction using various solvents with different polarity index. Among the extracts, the ethyl acetate portions were subjected to various pharmacological studies. Through this study, evaluate the antibacterial activities of extracted compounds and their nano forms. Methods Single compounds were obtained in each extract, which were characterized by UV, IR, and NMR and LC-MS techniques. Nanoporous materials of the same type were prepared by solvothermal and sonication methods. The obtained nanoporous materials were then impregnated on chitosan and their pharmacological evaluation was carried out. Results All compounds isolated from the plants show a minimum assay of antibacterial properties in their nanoforms. A better performance was obtained for Coleus zeylanicus Lour . Discussion Selected compounds from plectranthus amboinicus, plectranthus rotundifolius and plectranthus zeylanicus were converted to their nanoforms. These nanoforms were studied by antibacterial analysis. Some of these nanoforms combined with chitosan and their antibacterial activity were studied. Extracted compound from plectranthus zeylanicus using ethyl acetate extract were selected for this study. Bioavailability, chemical potential, nutritive contents and efficiency are the implications of these Compounds. Sustainability, specificity, reproducibility, and solubility of prepared compounds are the limitations of this study. Conclusion Chitosan impregnated Coleus zeylanicus Lour had high antibacterial potential against E.coli. growth.

&lt;p&gt; &lt;/p&gt;&lt;p&gt; &lt;/p&gt;&lt;p&gt;&lt;bold&gt;Objective:&lt;/bold&gt; To evaluate the acute effects of chewing fresh &lt;italic&gt;Plectranthus amboinicus&lt;/italic&gt; leaves on oral health parameters, including salivary microbiology, physicochemical properties, and clinicalindices. &lt;bold&gt;Materials and Methods:&lt;/bold&gt; A single-blind, randomized, controlled, crossover pilot study was conducted with 25 healthy adults. Participants underwent two interventions on separate days: chewing fresh &lt;italic&gt;P. amboinicus&lt;/italic&gt; leaves (experimental) or a tasteless cellulose gum (control) for 5 minutes. Salivary levels of &lt;italic&gt;Streptococcus mutans&lt;/italic&gt; and &lt;italic&gt;Candida albicans&lt;/italic&gt;, pH, flow rate, total antioxidant capacity (TAC), Plaque Index (PI), Gingival Index (GI), and volatile sulfur compounds (VSCs) were measured at baseline, immediately post-intervention (T1), and at 120 minutes (T3). Acceptability was recorded via questionnaire. &lt;bold&gt;Results:&lt;/bold&gt; Chewing &lt;italic&gt;P. amboinicus&lt;/italic&gt; resulted in a significantly greater and sustained reduction in salivary &lt;italic&gt;S. mutans&lt;/italic&gt; and &lt;italic&gt;C. albicans&lt;/italic&gt; counts compared to control (p&amp;lt;0.001), with effects persisting at T3. It induced a significantly stronger increase in salivary pH (to 7.65 ± 0.28 at T1 vs. 7.10 ± 0.25 for control, p&amp;lt;0.001) and boosted TAC at T3 (p&amp;lt;0.001). Significant reductions in PI and VSCs (68% reduction at T1) were also observed uniquely with &lt;italic&gt;P. amboinicus&lt;/italic&gt; (p&amp;lt;0.01). The intervention was rated as highly acceptable (92%). &lt;bold&gt;Conclusion:&lt;/bold&gt; Chewing fresh &lt;italic&gt;Plectranthus amboinicus&lt;/italic&gt; leaves provides immediate, multi-mechanistic oral health benefits—including antimicrobial, alkalinizing, antioxidant, anti-plaque, and anti-malodor effects—significantly surpassing mechanical chewing alone. This validates its traditional use and positions it as a promising natural adjunct for oral hygiene.&lt;/p&gt;&lt;p&gt; &lt;/p&gt;