Background <p>Head and Neck Squamous Cell Carcinoma (HNSCC) is a highly aggressive malignancy with significant morbidity and mortality. Tumour progression is influenced not only by cancer cells but also by the tumour microenvironment, including stromal characteristics. p16, a tumour suppressor gene and surrogate marker for HPV infection, plays a crucial role in HNSCC pathogenesis. This study aims to evaluate the correlation between p16 expression and fibrotic cancer stroma in HNSCC and its potential prognostic significance.</p> Materials and methods <p>A cross-sectional observational study was conducted on 87 histopathologically confirmed HNSCC specimens from a tertiary care hospital. Fibrotic cancer stroma was categorised into mature, intermediate and immature. Immunohistochemical staining for p16 expression was performed and classified according to the CAP 2018 protocol. The association between p16 expression and stromal patterns was statistically analysed using appropriate tests, with a significance threshold of <i>p</i> &lt; 0.05.</p> Results <p>Among 87 HNSCC cases, 80.46% were p16-negative, 14.94% were p16-positive, and 4.6% were equivocal. Stromal classification showed 40% immature, 37.5% intermediate, and 22.5% mature stroma. A significant association was found between p16 expression and stroma type (<i>p</i> = 0.034). Among p16-positive cases, 69.23% had intermediate stroma, 30.77% had mature stroma, and none exhibited immature stroma.</p> Conclusion <p>This study explores the significant correlation between p16 expression and stromal composition in HNSCC. The stromal characteristics may impact tumor behaviour in p16-positive HNSCC. Gaining insights into this relationship could enhance our understanding of tumor progression and prognosis.</p>