Assessment of In vitro Anti-Inflammatory Activity: A Comprehensive Review of Methods, Advantages, and Limitations

Abstract

Aims: Inflammation is a vital biological response that plays an important role during tissue repair and defense against infections. A range of transcription factors (including NF-KB and AP-1), enzymes (Cycooxygenase-2 (COX-2) and 5-Lipoxygenase (5-LOX) mediates this process. The dysregulation of inflammation can lead to chronic diseases such as atherosclerosis, cardiovascular disorders, and rheumatoid arthritis. Corticosteroids and Non-steroidal anti-inflammatory drugs (NSAIDs) are often used as anti-inflammatory drugs. Despite their effectiveness, the prolonged use of these drugs comes with various side effects. Therefore, there is a growing demand for safer but effective anti-inflammatory drugs. Plants and plant-based products have a rich history for their medicinal properties. Hence, medicinal plants can be a strong alternative for developing new anti-inflammatory drugs. Materials and Methods: In vitro assays are frequently utilized for the initial screening of natural products for their pharmacological properties as they are cost-effective, direct, and time-efficient. Protein denaturation inhibition assays, membrane stabilization assays, enzyme inhibition (COX/LOX), and nitric oxide assays are commonly used to evaluate the anti-inflammatory properties of drugs. Conclusion:  This review explores frequently utilized In vitro assays to evaluate anti-inflammatory activity, outlining their principles, methodologies, advantages, and limitations. It aims to help researchers choose appropriate techniques for drug discovery and the development of anti-inflammatory medications. However, the inability of In vitro models to fully replicate the complexity of living organism responses remains a significant limitation.