Neuro-immunity in stress-related oral ulcerations: A fractal analysis

By testing archival paraffinized biopsy blocks obtained from the oral pathology library with immunohistochemistry, we tested the hypothesis that substantial alterations are demonstrable in the cross-talk between sympathetic (VMAT) and para-sympathetic innervation (VAchT), and resident CD3+ T cells in the mucosa from oral lichen planus (OLP), compared to recurrent aphthous stomatitis (RAS) and control biopsies. We quantified fractal dimension and Euclidean dimension of CD3+ cells between the two pathologies, and across the set of CD3+ cells proximal to the vesicles of monoamines transport (VMAT)+ or the vesicles of acetylcholine transport (VAchT)+ innervation, compared to cells relatively distal to the nerve endings. The data show exquisite organization of the punctuate sympathetic and para-sympathetic staining about the resident CD3+ T cells in the OLP lesions, but not in the aphthous lesions or in control mucosa. Fractal analysis reveals that aphthous lesions are characterized by CD3+ T cells of larger size (Euclidean dimensional map), compared to control mucosa. CD3+ T cells in OLP lesions are also found to be significantly larger than those found in control lesions, when they are not proximal to sympathetic or para-sympathetic vesicles. The membrane of CD3+ T cells is overall more complex (fractal dimension) in aphthous lesions, compared to control sections. A similar trend is apparent, albeit not statistically significant, in CD3+ T cells resident in OLP lesions, whether or not they are located proximal to nerve endings. An overall decrease in the ratio of fractal dimension-over-topological dimension was also observed across the pathological lesions, compared to control. Taken together, these data indicate that as CD3+ T cells grow larger in the pathological conditions, they, in effect, stretch their plasma membrane, and that the cells may be at different stage of the cell cycle, relative to their position vis a vis nerve endings. Because fractal analysis is performed on individual cells, it has the potential of being developed in a novel diagnostic test, as well as a prognostic tool for monitoring the etiology and the course of treatment at the individual cellular level. Our findings also open new frontiers of fundamental, clinical and translational biosciences of OLP.