Background: Extracellular matrix is predominantly made of collagen that shows variation in structure and nature in different pathologies. There are overlapping histopathological features in dentigerous cyst (DC), normal dental follicle (NDF), and hyperplastic dental follicle (HDF), leading to diagnostic ambiguity. The distinction between the three lesions is important as they vary in biologic behavior. The efficacy of Masson's trichome (MT) and picrosirius red (PR) to distinguish dental follicles from DC has been minimally investigated. Aim: To determine the nature of collagen fibers in DC, NDF, and HDF using MT and PR stains, and to estimate the staining efficacy. Methodology: A total of 30 histopathologically diagnosed archival tissue sections each of NDF, HDF, and DC were chosen and subjected to routine and special stains, namely MT and PR. The resultant data was analyzed statistically using the chi-squared test. Results: The arrangement and orientation of collagen fibers differed significantly between NDF, HDF, and DC. The significant difference in the staining efficacy with respect to orientation, organization, arrangement, differentiation, and specificity suggests that PR is superior to MT and hematoxylin and eosin (H and E) in the detection of collagen as it has the potential to differentiate mature and immature collagen fibers. Conclusion: Picrosirius red (PR) and MT can be used to differentiate NDF, HDF, and DC, suggesting their role in determining the behavior of these lesions.