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The information of biomechanical properties is crucial in the study of biological tissue and its clinical relevance. 3mm x 3mm free gingival human tissues was taken using disposable punch biopsy (Accu sharp blade, India) and stored in 0°C Freezer. The sample was sectioned to a thickness of 10μm using high profile microtome blade (Leica 818, Germany) and cryostat (Leica CM1850UV, United Kingdom). The sample was analysed using Atomic Force Microscope (Nanowizard® 3, JPK Instruments, Germany) at room atmosphere. The collagen fibrils of the free gingival tissues appeared to be stacked in basket weave like structure. The mean value of free gingival collagen fibrils width and the length of D-band were 106.71±11.18nm and 65.82 ± 3.04nm respectively. The Young’s modulus of collagen fibrils for human free gingival tissue at overlap region was 212.88 ± 242.58 MPa, whereas at the gap region was 207.00 ± 230.71 MPa. Within the limitation of the study, the collagen fibrils appeared to be stacked in basket weavelike structure. The length and width of the collagen fibril were similar to the values investigated using other techniques. There was significant linear relationship between Young’s modulus of overlap and gap regions.