3D imaging of central centrifugal cicatricial alopecia treated with 6% topical gabapentin demonstrates re-innervation of the epidermis

Summary: Abstract Body: To evaluate the effect of 12 weeks of twice daily 6% topical gabapentin solution, we used hybrid open-top light sheet (OTLS) microscopy to analyze 3D spatial organization and cutaneous innervation in a patient with central centrifugal cicatricial alopecia (CCCA). In this qualitative study, pre- and post-treatment biopsy specimens from affected and unaffected scalp of a patient with CCCA were stained with TO-PRO-3, PGP9.5, and CD45, optically cleared with a modified iDISCO protocol, and imaged with a hybrid OTLS microscope at varying resolutions. OTLS microscope imaging of scalp tissue produced a detailed 3D assessment of intact hair follicle structures and cutaneous innervation. This technology rapidly imaged overall tissue architecture at lower resolution (2 µm/pixel), with deeper analysis of pathologic features at higher resolution (0.17 µm/pixel). A clear increase in epidermal nerve fiber density in the affected scalp after topical gabapentin treatment was observed. Results demonstrated that 6% topical gabapentin solution was potentially linked with improved peripheral cutaneous innervation of CCCA-affected scalp tissue. 3D spatial imaging with OTLS microscopy enabled visualization and quantification of structures that are typically poorly visualized with 2D histopathologic evaluation, such as nerve fibers innervating the hair follicle and epidermis. Application of OTLS technology to samples with CCCA provided new insights into pathologic changes in CCCA and neuronal remodeling with 6% topical gabapentin. This technology addresses sampling bias inherent in traditional 2D slide-based methods by capturing the entire sample volume and by providing a comprehensive view of spatial structures. OTLS microscopy holds potential to enhance scalp biopsy analysis, offering new opportunities for histopathologic evaluation of cicatricial alopecias and correlation with trichoscopic findings. Divya R. Alley¹, Shikhar Dhingra², Sara Bohjanen¹, Irmina Wallander¹, Maria Hordinsky¹ 1. Department of Dermatology, University of Minnesota Medical School, Minneapolis, MN, United States. 2. Alpenglow Biosciences, Seattle, WA, United States. Bioinformatics, Computational Biology, and Imaging