<p>Hair follicle stem cells (HFSCs) have been studied for their role in developmental biology, hair cycling and transplantation. However, recent research reframes the hair follicle as a heterogeneous regenerative ecosystem capable of responding to injury cues and contributing to tissue repair beyond trichology. Despite advances, our understanding of HFSC physiology and therapeutic applications remains fragmented. Many reviews fail to bridge the gap between mechanistic biology and translatable therapeutic modalities, often overlooking critical aspects such as stemness-preserving culture systems, biomaterial-assisted delivery, and the specific influence of distinct wound microenvironments on HFSC fate. This review aims to delineate the current landscape of HFSC research, synthesizing evidence for their therapeutic potential beyond hair regeneration and discussing translational bottlenecks. This study summarizes core physiological functions, including injury-driven re-epithelialization and melanocyte stem cell-coupled re-pigmentation. We synthesize evidence for HFSC-based therapies in acute/chronic wounds (including diabetic and venous ulcers), burns, vitiligo, and exploratory applications in diabetes and ocular reconstruction. Furthermore, we introduce strategies for stemness-preserving expansion, preconditioning, and bioengineering delivery systems. This review provides a reference for advancing HFSC-based therapies toward reproducible, clinically viable interventions by integrating physiological insights with engineering solutions to overcome current translational barriers.</p>