Treatment and management of alopecia are highly determined by an accurate diagnosis, which can be challenging due to the lack of methods to properly visualize hair follicles. Current standard diagnosis is based on dermoscopy for non-scarring alopecia and scalp biopsy for scarring types of alopecia. Dermoscopy can be inconclusive, while biopsy is a painful procedure. In this study, we used a clinical tomograph based on multiphoton microscopy (MPM) to non-invasively image the scalp of 5 healthy subjects and of 12 patients affected by non-scarring alopecia (androgenetic and areata) and scarring alopecia (frontal fibrosing). MPM is capable of non-invasive in vivo imaging of follicular structures in human scalp including hair shafts, hair follicles and sebaceous glands via two-photon excited fluorescence (TPEF) from keratin and NADH/FAD and of the papillary dermis surrounding the hair follicles through second harmonic generation (SHG) from collagen and TPEF from elastin fibers. In normal and non-scarring alopecia patients, MPM often identified presence of sebaceous glands associated with hair follicles, while MPM images of scarring alopecia were characterized by miniaturization of hair follicles as well as by presence of macrophages and lymphocytes surrounding hair follicles. A quantitative analysis involving measurement of hair follicle diameter sizes showed they were significantly smaller in scarring comparing to non-scarring alopecia patients and to normal scalp subjects (p < 0.043). This study shows, in a limited number of patients, that MPM imaging can non-invasively identify morphological features that distinguish scarring from non-scarring alopecia.
Androgenic alopecia(AA), commonly known as pattern baldness, initially manifests as a receding or thinning hairline in men and women respectively. While the condition may be apparent at a later stage of development, it is routinely cumbersome to diagnose in its early stages. The current gold standard for AA diagnosis is trichoscopy of the scalp and the more invasive punch biopsy when trichoscopy yields insufficient insight towards the cause of hair loss. Trichoscopy, a magnified image of the scalp, can provide a superficial understanding of the hair diameter as well as scalp discoloration around the follicular opening. Histopathological analysis of hair follicles provides information such as hair follicle density, and the diameter of the hair shaft and follicle used to determine the extent of alopecia. Since the information provided by trichoscopy is somewhat limited and punch biopsies are invasive, physicians cannot properly assess multiple areas of the scalp to provide proper diagnosis of androgenic alopecia. Optical Coherence Tomography(OCT), a non-invasive, non-ionizing tomographic imaging technique, can rapidly provide near histopathological cross sectional 3D images. This technique has been identified as a plausible candidate for in-vivo assessment of hair loss. Through this pilot study, we have prospectively enrolled eight patients undergoing varying degrees of hair loss to assess the efficacy of OCT to detect differences in hair shaft and follicle anatomy. Our preliminary results show the capability of OCT to non-invasively resolve hair follicle density, hair shaft and follicle diameters between patients at varying stages of hair loss.
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