Inflammation is a natural defense mechanism of the body in response to harmful stimuli such as pathogens, injuries, or irritants. However, chronic inflammation is associated with many disease including arthritis, asthma, and cardiovascular disorders. Although synthetic anti-inflammatory drugs are widely used, they often have adverse effects and limitations. Therefore, medicinal plants have gained attention due to their traditional use and natural therapeutic potential. This review focuses on both in vitro and in vivo screening methods used to evaluated the anti-inflammatory activity of various medicinal plants. In vitro methods such as membrane stabilization, protein denaturation and nitric-oxide inhibition provide rapid and cost-effective preliminary results. In vivo models, including carrageenan induced paw edema and formalin induced paw edema offer a more comprehensive evaluation in living systems. This article summarize the finding of several studies that have 0the development of safer and effective anti-inflammatory agents (Phanse, 2012).
Abstract
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Keywords
Anti-inflammatory activity
Herbal Medicine
In vitro model
In vivo model
Medicinal Plants
INTRODUCTION
Inflammation is a complex biological response of the body’s immune system to harmful stimuli such as pathogens, damaged cells, or irritants. While acute inflammation is a protective mechanism essential for healing, chronic inflammation is a pathological condition implicated in numerous diseases, including arthritis, cancer, cardiovascular disorders, and neurodegenerative conditions. Conventional anti-inflammatory drugs, including non-steroidal anti-inflammatory drugs and corticosteroids, are effective in managing symptoms but are often associated with significant side effects such as gastrointestinal bleeding, renal toxicity, and cardiovascular risks when used long-term (Yada et al., n.d.).
Inflammation is a vital defense mechanism that protects the body against infections, injuries, and foreign substances. It involves a coordinated cascade of molecular and cellular events that aim to eliminate the initial cause of cell injury and initiate tissue repair. While acute inflammation is typically short-lived and beneficial, chronic inflammation can contribute to the development of several pathological conditions including rheumatoid arthritis, asthma, inflammatory bowel disease, cardiovascular disorders, and neurodegenerative diseases (Oladejo and Akingbade, 2024; Yada et al., n.d.).
This review aims to explore the current advancements in nano formulated anti-inflammatory agents, emphasizing the relevance and methodology of both in vitro and in vivo assessments. It also highlights recent studies, models used, and the potential of nano formulations in overcoming the limitations of conventional anti-inflammatory therapies.
The evaluation of nano formulated drugs requires a dual approach-in vitro studies help understand the cellular mechanisms, cytotoxicity, and anti-inflammatory potential, while in vivo models are crucial to assess pharmacokinetics, efficacy, and safety in living systems. Together, these assessments provide a comprehensive understanding of the therapeutic potential of nano formulated anti-inflammatory agents (Oladejo and Akingbade, 2024).
Inflammation can be categorized into
The goal of acute inflammation is to remove the cause of injury, eliminate dead cells, and initiate tissue repair (Murt Huza and Manjunatha, 2018a).
Chronic inflammation
Chronic inflammation is a prolonged and persistent [inflammatory response that occurs when the acute phase fails to eliminate the cause of injury or when the harmful stimulus continues over time.
Although initially protective, chronic inflammation can lead to tissue destruction, fibrosis, and organ dysfunction (Murt Huza and Manjunatha, 2018a).
Important of inflammation
Why Anti-Inflammatory Drug Are Needed
Inflammation, though a natural and protective response of the body, can become harmful when it is excessive, uncontrolled, or chronic. In such conditions, the body immune response starts damaging healthy tissues, leading to pain, swelling, organ dysfunction, and chronic disease. To manage this, anti-inflammatory drugs are necessary (Xie and Huang, 2024).
Limitations of conventional drugs
Despite their importance, many conventional anti-inflammatory drugs (e.g., NSAIDs, corticosteroids) have side effects such as:
Pathophysiology of Inflammation
Inflammation is a protective and coordinated biological response of the body’s immune system to harmful stimuli such as pathogens, damaged cells, toxins, or physical injury. Its main objective is to eliminate the causative agent, remove dead or damaged tissue, and initiate tissue repair and regeneration. However, if uncontrolled or unresolved, inflammation itself may cause tissue injury and contribute to various chronic diseases.
The pathophysiological process of inflammation involves a sequence of events that include vascular changes, recruitment of immune cells, and release of chemical mediators (Li, Chen, and Zhang, 2023).
Phase of Inflammation
Chemical mediators of inflammation
Chemical mediators are central to the initiation, amplification, and resolution of the inflammatory response. They can be derived from.
Need for Herbal Alternatives in Inflammation Management
Inflammation is a critical component of the body defense system however, when prolonged or dysregulated, it contributes to chronic conditions such as rheumatoid arthritis, inflammatory bowel disease, asthma, cancer, and metabolic syndromes. While modern pharmacological agents like non-steroidal anti-inflammatory drugs and corticosteroids are effective in managing inflammation, their long-term use is frequently associated with serious adverse effects, including gastrointestinal ulcers, renal impairment, hypertension, and increased risk of infections (Mohseni et al., 2020).
Limitations of Conventional Anti-Inflammatory Drugs
Herbal Alternatives: A Safer Option
Evidence of Efficacy
Several medicinal plants such as Curcuma longa (turmeric), Zingiber officinale (ginger), Boswellia serrata (salai guggul), and with Ania somniferous (ashwagandha) have shown significant to standard drugs like indomethacin or diclofenac (Jabn, 2017; Jayasuriya et al., 2017).
In vitro Models for Anti-Inflammatory Screening
In vitro screening methods are widely used in the early stages of drug discovery to screen and evaluate the anti-inflammation potential of medical plants. These models are simple, cost-effective, and a to allow for high-throughput screening. They primarily asses the ability of plant extracts or compounds to inhibit inflammation related biological processes at the cellular or biochemical level (Yada et al., n.d.).
Human Red Blood Cell (HRBC) Membrane stabilization Assay
Plant Used: Abutilon indicum Linn.
Family: Malvaceous.
Part Used: Leaves.
Solvent: Ethanol-4.5% wt/wt, Chloroform-0.45% wt/wt and distilled water-3.7% wt/wt.
Dose: 50 mg/100 mL.
Chemical Used: Diclofenac, phosphate buffer, HRBC suspension.
Principle
During inflammation, lysosomal enzymes are released, which can cause further tissue damage. The HRBC membrane is similar to the lysosomal membrane, and its stabilization indicates anti-inflammatory activity (Daram et al., 2021).
Procedures
The HRBC membrane stabilization has been used as method to study the anti-inflammation activity. Blood was collected from healthy volunteer. The collected blood was mixed with equal volume of sterilized Al sever solution (2% dextrose, 0.8% sodium citrate acid and 0.42% sodium chloride in water). The blood was centrifuged at 300 r.p.m. and paced cell were washed with isohaline. The assay mixture contained the drug. 1 ml of phosphate buffer (0.15 m, pH), 2 mL of hyposaline (0.36%) and 0.5 mL of HRBC suspension. Diclofenac was used as reference drug. Instead of hyposaline 2 mL of distilled water was used in the control. All the assay mixture were incubated at 37ºC for 30 min and centrifuged. The hemoglobin content in the supernatant solution was estimated using spectrophotometer at 560 nm. The percentage hemolysis was calculated by assuming the hemolysis produced in presence of distilled water of as 100% (Daram et al., 2021).
Protein Denaturation Assay
Plant Used: Curcuma longa Linn.
Family: Zingiberene.
Part used: Rhizomes.
Solvent: Ethanol, 70% wt/wt.
Chemical Used: Bovine Serum Albumin (BSA), Hydrochloride Acid (HCl), Indomethacin.
Principal
Inflammation often involves denaturation of protein due to heat or stress. Compounds that prevent protein Denaturation can potentially reduce inflammation. In this assay, BSA is used as a model protein, and inhibition of its denaturation by the test extract indicates anti- inflammation activity (Ewig et al., 2025).
Procedures
Prepare 1% aqueous solution of BSA. Mix 0.45 mL of BSA solution with 0.05 mL of plant extract (various concentration). Adjust pH to 6.3 using 1 N HCl. Incubate the sample at 37ºC for 5 min. Cool the sample and measure absorbance at 660 nm using UV spectrophotometer. Calculation % inhibition of protein denaturation using the formula (Oladejo et al., 2024).
% Inhibition = Absorbance Control-Absorbance Sample×100
Absorbance Control
Albumin Denaturation Assay
Plant Used: Abutilon indicum Linn.
Family: Malvaceous.
Part Used: Leaves.
Solvent: Ethanol, 70% wt/wt.
Chemical Used: Fresh egg albumin (BSA as alternative), Diclofenac (standard), Phosphate buffer saline, HCl (for pH adjustment).
Principle
Heat or chemical stress causes denaturation of albumin, denatured albumin increase turbidity. Substances that prevent or reduce this denaturation show anti-inflammatory potential. This assay measures inhibition of albumin denaturation by the test sample spectrophotometrically (El-Shivani and Eid, 2019).
Procedure
Prepare albumin solution if using egg albumin dilute fresh egg white with distilled water to obtain 1% (wt/wt) albumin solution. Or prepare 1% BSA in distilled water as alternative.
Prepare test extract solution at required concentration (example: 10, 50, 100, 200 g/mL) in ethanol or phosphate buffer saline. Prepare standard solution diclofenac at appropriate concentration.
Prepare a control containing albumin + buffer (no extract). Prepare a blank for each sample (same as sample but without albumin) to zero instrument for solvent absorbance. Mixing in labeled test tube, add 0.5 mL of test extract (or standard/control solvent). 0.45 mL of albumin solution (1%). 0.05 mL of phosphate buffer (to adjust volume/Ph). For control 0.5 mL solvent+0.45 mL albumin + 0.05 mL buffer.
For blank 0.5 mL test extract+0.5 mL buffer. Incubate the mixtures at 37ºC for 15-20 min. Heat the tube at 70ºC for 5 min. Allow to cool to room temperature (Murt Huza and Manjunatha, 2018b).
% Inhibition = Absorbance control-Absorbance sample×100
Human keratinocyte HaCaT
Family: Zingiberene.
Plant used: Acorus calamus L.
Part Used: Leaves.
Solvent: Water.
Dose: 50 mg/kg.
Route of administration: intraperitoneally,
Chemical Used: Immunofluorescence staining, polyinosine, polypyridyl acid.
Principle
Human keratinocytes, such as the HaCaT cell line, are skin derived epithelial cells that play a crucial role in inflammation and immune responses. In an in vitro anti inflammation assay, test compound such as plant extract or drug are applied to keratinocyte cultures before or after inducing inflammation. The ability of the compound to reduce inflammatory. The ability of the compound to reduce inflammatory mediator level, inhibit COX/LOX enzymes, or down regulate inflammatory gene expression is measured. This model mimics skin inflammation and allows screening of anti-inflammatory potential at the cellular level without using animal (Kedi et al., 2018).
Procedure
HaCaT cells treated with polyinosine: Polypyridyl acid and peptidoglycan induced the inflammatory reaction. The anti-inflammatory activities of ACL were investigated using RT-PCR, ELISH assay, western blotting, and immune fluorescence staining (Kedi et al., 2018).
Lipoxygenase Inhibition Assay
Plant Name: Alepidea amatymbica.
Family: Apiaceae.
Part Used: Roots.
Solvent for Extraction: Acetone.
Chemical Used: Soybean lipoxygenase enzyme (EC 1.13.11.12), sodium phosphate buffer 0.1 m, pH 6.8 (or pH 7.0 as per protocol), linoleic acid substrate.
Principle
Lipoxygenase inhibition assay involves measuring the enzymes ability to catalyze the oxidation of unsaturated fatty acid, typically linoleic acid, into a hydroperoxide. Inhibitors of LOX will reduce this reaction, which is then quantified through various methods, often involving a color change. Lipoxygenases are present in the human body and play an important role in the stimulation of inflammatory reactions (Morass et al., n.d.).
Procedure
Buffer 0.1 m sodium phosphate, pH 6.8. Enzyme dilute soybean LOX to working concentration (example: 1,000 U/mL stock → dilute to provide ~10 mL/well optimize enzyme amount so the control gives linear increase at 234 nm). Substrate linoleic acid 10 mm stock. Prepare fresh and keep on ice. 160 mL sodium phosphate buffer (0.1 m, pH 6.8)10 mL lipoxygenase enzyme solution 10 mL test sample solution (or 10 mL buffer for control mL standard for positive control). Pre-incubate enzyme + sample mixture at 25ºC for 5 min. Record absorbance increase at 234 nm every 15-30 sec for 3-5 min or measure initial rate. Use control wells no inhibitor to get enzyme activity (Morass et al., n.d.).
DPPH assay
Plant Name: Curcuma longa.
Family: Zingiberaceae.
Part used: Rhizomes.
Solvent: 70% ethanol.
Chemical: DPPH (0.1 mm in methanol), methanol (HPLC/AR grade), ascorbic acid (standard), phosphate buffer, microplate/UV.
Principle
DPPH (2,2-Diphenyl-1-Picrylhydrazyl) assay is an antioxidant activity test based on a color change reaction involving a stable, violet colored free radical. When DPPH reacts with an antioxidant, it becomes a reduced, pale- yellow molecule. The degree of color change is measured using a spectrophotometer to quantify the sample antioxidant capacity (Peiris et al., 2025).
Procedure
This experimental procedure in an ethanol solution of 2,2-diphenyl-1-picrylhydrazyl radical (final concentration was 1.0×10−4 m), test extract at different concentration were added. The reaction mixture were shaken vigorously and then kept in the dark for 30 min. The absorbance of the resulting solutions was measured in 1 cm cuvettes, using a PerkinElmer Lambda 40 UV/visible spectrophotometer at 517 nm, against blank without DPPH. Decreasing of DPPH solution absorbance indicates an increase of DPPH radical scavenging activity. This activity is given as %DPPH radical scavenging that calculated in the equation.
In vivo Models for Anti-Inflammatory Screening
In vivo model are essential tools for evaluating the anti-inflammatory potential of plant extracts and synthetic compounds under physiological conditions. They help in understanding the integrated response of cells, tissues, and organ systems to inflammatory stimuli. Both acute and chronic inflammation can be studied using various models, each specific pathways and mediators involved in the inflammatory process (Chen et al., 2018).
Carrageenan-Induced Paw Edema
Plant used: Hydrocotyle Batrachium.
Family: Apiaceae.
Part Used: Whole plant.
Common Name: Marsh pennywort.
Solvent: Water.
Dose: 500 and 100 mg/kg.
Route of administration: Orally.
Chemical used: Indomethacin.
Principle
Carrageenan hind-paw injection causes biphasic acute edema. The test drug/extract reduces paw swelling and control. These models mimic acute and chronic inflammatory condition and measure parameter such as edema formation, granuloma development, or vascular permeability. The principle is based on comparing the inflammatory response in treated animals against controls, allowing the determination of anti-inflammatory potential (Umar et al., 2010).
Procedure
The anti-inflammatory activity of HBW (Hydrocotyle batrachium Hance) was determined by the carrageenan induced edema test. Male and mice were randomly assigned to five groups and then fasted with free access to water for 24 hr before the experiment. Fifty microliters of a 1% suspension of carrageenan in saline, which had been prepared 30 min before each experiments, was injected into the plantar side of the right hind paw of the mice. After 60 min, HBW at dose of 100, 500 and 1,000 mg kg−m were administered orally, and after 90 min, indomethacin at the dose of 10 mg kg−m was administered an intraperitoneal route after the carrageenan treatment. Paw volume was measured before carrageenan injection and at 60, 120, 180, 240, 300, 360 min after the administration of the indomethacin agent using a plethysmometer (Lantz et al., 2005).
Cotton Pellet-Induced Granuloma in Rats
Plant Used: Cássia fistula Linn.
Family: Caesalpinaceae.
Part used: Bark.
Common Name: Amalltas.
Solvent: Water and methanol.
Dose: 2,000 mg/kg.
Rote of administration: Orally.
Chemical Used: Diclofenac sodium.
Principle
Subcutaneous sterile cotton pellets induce proliferative chronic inflammation (fibroblast proliferation and collagen deposition). Reduced dry granuloma weight indicates anti-proliferative/anti-inflammatory effect. The anti-inflammatory potential of a test compound is assessed by its ability to reduce the dry weight of the granuloma, which reflects inhibition of protein and collagen synthesis during chronic inflammation (Cheng et al., 2016).
Procedure
Wistar albino rats (170-200 g) of either six divided into 4 groups of 6 animal in each group. Cotton pellets weighing 30+1 mg were autoclave and implanted subcutaneously into both sides of the groin region of each rat, Group1 served as control and received the vehicle. The extracts CFA and CFM at concentration 500 mg/kg orally for Group 2 and Group 3 animals for same period. On the eighth day the animals were sacrified and the pellets together with the granuloma tissues were carefully removed, dried in an oven at 60ºC, weighed and compared with control. Diclofenac sodium, 5 mg kg− was used as standard (Gonfa et al., 2023).
Formalin Induced Paw Edema in rats
Plants used: Crocus sativus Linn.
Family: Iridaceae.
Part Used: Powder of stigma and petal.
Common Name: Saffron.
Solvent: Water and ethanol.
Dose: 0.8 and 2 g/kg.
Route of administration: Intraperitoneally.
Chemical used: Diclofenac.
Principle
The formalin induced paw edema model evaluates both neurogenic and inflammatory pain responses. Subcutaneous injection of dilute formalin into a rat hind paw produces biphasic edema and pain. The anti-inflammation activity of a drug is measured by its ability to reduce paw swelling (volume or thickness). In the late phase, reflecting inhibition of mediator release and vascular permeability (Al-Khayri et al., 2022).
Procedure
Rats were divided into groups of six. The inflammation was produced by subaponeurotic injection of 0.1 of 2% formaldehyde in the right hind paw of the first and third day. The animals were treated daily with the extracts or diclofenac intraperitoneally for 10 days. The daily changes in paw size were measured by wrapping a piece of cotton threads round the paw and measuring the circumference with a meter rule (Al-Khayri et al., 2022).
Xylene Induced ear edema
Plant Used: Crocus sativus Linn.
Family: Iridaceae.
Part Used: Powder of stigma and petal.
Common Name: Saffron.
Solvent: Water and ethanol.
Dose: 0.8 and 2 g/kg.
Rote of administration: Intraperitoneally.
Chemical used: Diclofenac, dexamethasone.
Principle
Xylene induced ear edema is the topical application of xylene to the pinna produces an acute irritant dermatitis leading to rapid vasodilation, increased vascular permeability, and neurogenic inflammation. This causes plasma extravasation and tissue fluid accumulation in the treated ear (Al-Khayri et al., 2022).
Procedure
Mice were divided into group of seven. 30 min after intraperitoneally. injection of the extract, diclofenac and dexamethasone, 0.03 mL of xylene was applied to the anterior and posterior surfaces of the right ear. The left ear was considered as control. 2 hr after xylene application mice were killed and both ears were removed. Circular section were taken using a cork borer with a diameter of 7 min, and weighed. The increased in weight caused by the irritant was measured by subtracting the weight of the untreated left ear section from that of the treated right ear section (Al-Khayri et al., 2022).
Cotton Oil-Induced Mouse Ear Edema
Plant Used: Anthrium cerrocampanense.
Family: Araneae.
Part used: Whole plant.
Common Name: Tailflower.
Solvent: Water, ethanol, dichloromethane.
Dose: 100 mg/kg.
Route of administration: Intraperitoneally.
Chemical Used: Indomethacin.
Principle
Croton oil induced ear edema in mice is topical application of croton oil on the mouse ear produce acute and sub acute inflammation, mainly through the release of arachidonic acid metabolites and proinflammatory cytokines. The anti-inflammatory activity of a drug is assessed by its ability to reduce the increase compared to control. This model is especially useful for evaluating topical anti-inflammatory agents (Al-Khayri et al., 2022).
Procedure
Cutaneous inflammation was induced to their immer surface of the right ear of anesthetized mice (100 mg kg−m ketamine hydrochloride, intraperitoneally) by application of 15 mL of acetone or acetone-water (1:1) solution contain 80 mg of croton oil as irritant (control animals). For treated animals, appropriate amounts of indomethacin or the tested extract were dissolved in the croton oil containing solution and applied as for controls. After 6 hr mice were sacrificed and a punch (5 mm of diameter) was excised from both ears. Inflammation was measured as edema formation and quantified by the weight difference between the treated and the untreated ear sample (Al-Khayri et al., 2022).
Phytoconstituents Responsible for Anti-Inflammatory Activity
Medicinal plants contain a wide range of bioactive phytochemical that contribute to their anti-inflammatory effects. These compounds act through different mechanisms such as inhibition of cyclooxygenase, lipoxygenase, suppression of proinflammatory cytokines and downregulation of NF-κB signaling pathways. Plant based phytoconstituents are knows for their potential sources of anti-inflammatory agents. Various studies reported the potential disease healing efficiency of medical plants. Medicinal plants are the primary source of medicines (Ansari et al., 2025).
Alkaloids
Example of Plants: Berberis vulgaris (Berberine), Colchicum autumnal (Colchicine).
Mechanism
Inhibit neutrophil migration, reduce cytokine production, and suppress NF-κB Activity.
Role
Help in reducing acute and chronic inflammation (Al-Khayri et al., 2022).
Flavonoids
Example of Plants: Allium cepa (Quercetin), Ginkgo biloba (Kaempferol), Citrus (Hesperidin).
Mechanism
Inhibit COX/LOX enzymes, scavenge free radicals, stabilize cells membranes and downregulate proinflammatory mediators.
Role
Provide strong antioxidant and anti-inflammatory protection (Ferraz et al., 2020).
Phenolic Compound
Example of plants: Curcuma longa (Curcumin), Vitis vinifera, Syzygies aromaticum
Mechanism
Suppress NF-κB activation, reduce oxidative stress, inhibit release of inflammatory mediators.
Role
Act as dual antioxidants and anti-inflammatory mediators (Ferraz et al., 2020).
Glycosides and saponins
Example of plants: with Ania somniferous, Panax ginseng (Ginsenosides).
Mechanism
Inhibit proinflammatory cytokines stabilizes lysosomal membranes and reduce vascular permeability.
Role
Effective in both acute and chronic inflammatory (Ferraz et al., 2020).
Limitations of Current Studies
Despite significant progress in evaluating the anti-inflammatory potential of medicinal plants and their formulations, several limitations restrict their translation into clinical use. Current anti-inflammatory research faces limitations related to the complexity of inflammation, the limitations of in vitro and in vivo models, and the challenges in translating research findings into effective and safe drugs. Specifically, while various in vitro and in vivo models exist, they may not fully replicate the complexity of the human inflammatory response (Winter et al., 1962).
Lack of clinical data
Most studies are restricted to in vitro and animal models, with very few clinical trails. This makes it difficult to confirm the real therapeutic potential in humans. Its is especially large scale a high-quality studies demonstrating efficacy and safety in humans, are often lacking for promising anti-inflammatory compounds, particularly those derived from natural sources (Itharat et al., 2024).
Poor standardization of plant extracts
Variation in extraction methods, solvents, and plant material leads to inconsistent results and reduced reproducibility. The complex nature extracts and the difficulty in isolating pure compounds make it challenging to identify and develop lead compounds (Itharat et al., 2024).
Variability in experimental models
Ethical consideration regarding animal welfare, along with inherent difference between animal model and human present challenges in using in vivo models for anti-inflammatory.
Difficulty in complexity of inflammation in in vitro models
In vitro cell culture models provide a simplified environment and struggle to replicate the intricate interplay of cells, tissues, and systems involved in the inflammatory response observed in living organisms.
Challenges in reproducibility
Difficulties in reproducing experimental results, even within the same laboratory, can limit the reliability of preclinical findings and impede their translation into clinical application.
Toxicity and Safety Concerns
‘Long-term safety profiles’ are ready studied. Some plants may contain toxic constituents or interact with conventional drugs, posing risk in clinical use (Oppong et al., 2024).
Future Prospects of Anti-Inflammatory
Despite the availability of numerous anti-inflammatory drugs, current therapies such as non-steroidal anti-inflammatory drugs and corticosteroids are often associated with severe side effects including gastrointestinal toxicity, cardiovascular complications, and immunosuppression. Therefore, there is a growing need to develop novel and safer anti-inflammatory agents. The future of anti-inflammatory therapy is expected to shift toward targeted, safer, and patient-specific strategies.
Safer Conventional Therapies
The development of next generation NSAIDs with improved selectivity of COX-2 while minimizing cardiovascular risk remains a major focus. Similarly, novel corticosteroid formulations that provide localized delivery (such as inhaled or topical steroids) aim to reduce systemic adverse effects while retaining therapeutic efficacy.
Biologics and Monoclonal Antibodies
Biologics targeting proinflammatory cytokines (e.g., TNF-α, IL-1β, IL-6) have revolutionized the management of autoimmune and chronic inflammatory disorders. Future research is directed toward next generation biologics that are safer, less immunogenic, and potentially available in oral formulations. Additionally, biosimilars and combination therapies are expected to increase accessibility and therapeutic success.
Nanomedicine and Targeted Delivery
Nanotechnology offers a promising approach to enhance drug delivery by enabling site-specific targeting of inflamed tissues. Nano formulations such as liposomes, polymeric nanoparticles, and dendrimers improve drug solubility, stability, and bioavailability while reducing systemic toxicity. Future nanomedicine may employ smart nanocarriers responsive to local stimuli (pH, temperature, enzymatic activity), allowing controlled and sustained release of anti-inflammatory agents.
Plant Based and Natural Anti-Inflammatory
Phytochemicals such as curcumin, quercetin, resveratrol, and Boswellia acids have demonstrated significant anti-inflammatory potential through multitarget mechanisms. The future focus lies in standardizing herbal extracts, conducting rigorous clinical validations, and overcoming limitations of poor bioavailability by formulating herbal nano drug delivery systems. Moreover, ethnopharmacological research continues to provide new leads for plants derived therapeutics
Gene and RNA Based Therapies
Advances in molecular biology have opened opportunities to modulates inflammation at the genetic level. RNA based drugs such as siRNA and miRNA can selectively silence proinflammatory mediators. Similarly, CRISPR-Cas9 gene editing technology holds potential for long-term modulation of inflammatory pathways, particularly in genetic or chronic inflammatory conditions.
Microbiome Targeted Therapies
The human gut microbiota has emerged as a crucial regulatory of inflammation and immune responses. Strategies to restore microbiome balance using probiotics, prebiotics, and postbiotics are being actively investigated. Fecal microbiota transplantation represent another 1 innovative approach for the treatment of inflammatory bowel disease and related disorders.
Personalized and Precision Medicine
Interindividual variability in drug response necessitates a move toward precision medicine. Biomarker based therapy can allow selection of the most effective drug for each patient, thereby improving treatment outcomes and minimizing side effects. The integration of artificial intelligence and computational drug discovery is also expected to accelerate the identification of novel anti-inflammatory agents.
Novel Mechanistic Targets
Emerging research highlights new pathways in inflammation beyond COX enzymes and cytokines. Inhibitors of the NLRP3 inflammasome, modulators of the JAK-STAT pathway, and cannabinoid receptor agonists are being explored as next-generation therapeutic options, particularly in neuroinflammatory and autoimmune conditions.
The detail literature survey was done on various plants which can be used in inflammation including their plant used, family, part used, common name, solvent of extraction, yield, dose, route of administration, chemical used, Procedure mechanism of action, and Conclusion was done. It is cleared from mechanism of action of these extract that almost all drugs work on the principle of inhibition of prostaglandin and other inflammatory mediators synthesis. Inflammation is a complex biological response that plays a central role in the pathogenesis of several acute and chronic diseases. Current synthetic anti-inflammatory drugs, though effective, are often limited by adverse effects, drug resistance, and long-term safety concerns. In this context, the exploration of medicinal plants and novel drug delivery systems, including nano formulations, offers promising alternatives. Evidence from both in vitro and in vivo studies highlights their ability to modulate inflammatory mediators, stabilize cell membranes, and attenuate tissue damage, thereby supporting their therapeutic relevance. Overall, medicinal plants and nano formulations represent a valuable frontier in anti-inflammatory drug discovery, with the potential to provide safer, effective, and sustainable alternatives for managing inflammation-related disorders (Winter et al., 1962).
CONCLUSION
Inflammation is a complex biological response that plays a crucial role in the pathogenesis of numerous acute and chronic diseases. Although conventional anti-inflammatory drugs such as NSAIDs and corticosteroids are effective, their prolonged use is often associated with significant adverse effects and safety concerns. This has led to increasing interest in alternative therapeutic approaches, particularly those derived from medicinal plants.
The present review comprehensively summarizes various in vitro and in vivo models used for evaluating anti-inflammatory activity. In vitro assays such as membrane stabilization, protein and albumin denaturation, lipoxygenase inhibition, DPPH assay, and cell-line-based studies provide rapid, cost-effective, and mechanistic insights into anti-inflammatory potential. In vivo models including carrageenan-induced paw edema, cotton pellet-induced granuloma, formalin-induced inflammation, and ear edema models offer a more integrated assessment under physiological conditions. Together, these models form a complementary framework for screening and validating potential anti-inflammatory agents.
Phytoconstituents such as flavonoids, alkaloids, phenolic compounds, glycosides, and saponins demonstrate significant anti-inflammatory effects through multiple mechanisms, including inhibition of cyclooxygenase and lipoxygenase pathways, suppression of pro-inflammatory cytokines, stabilization of lysosomal membranes, and modulation of NF-κB signaling. Furthermore, emerging approaches such as nano-formulations enhance drug delivery, improve bioavailability, and reduce systemic toxicity, representing a promising advancement in anti-inflammatory therapy.
Despite encouraging preclinical findings, limitations such as lack of standardization, insufficient clinical trials, reproducibility challenges, and safety concerns must be addressed before widespread clinical translation. Future research should focus on rigorous clinical validation, improved extract standardization, advanced drug delivery systems, and precision-based therapeutic strategies.
Overall, medicinal plants and nano-based formulations represent a promising and sustainable frontier in anti-inflammatory drug discovery, with the potential to provide safer, more effective, and targeted therapies for inflammation-related disorders.
Conflict of Interest
The authors declare that there is no conflict of interest.
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