Human olfactory-derived neural progenitors diminish locomotory deficits following spinal cord contusion injury

摘要

Objective: To investigate the therapeutic utility of human adult olfactory epithelial-derived neural progenitors (hONPs) as an autologous cell-based treatment for spinal cord injuries (SCIs).Design: To characterize and compare populations of hONPs obtained from cyrostor-age. Enzyme-linked immunosorbent assay (ELISA) was used to evaluate in vitro synthesis and release of neurotrophins (brain-derived neurotrophic factor, nerve growth factor, cilliary neurotrophic factor (CTNF), neurotrophin 3, and glial-derived neurotrophic factor) as well as fibroblast growth factor and vascular endothelial growth factor. To determine the efficacy of hONP engraftment on rats with computer-controlled moderately severe T9 contusions, the improvement compared to fibroblast engrafted and nonengrafted controls was evaluated. Studies employing behavioral, biochemical, electrophysiological, and morphological techniques were applied to SCI animals and isolated cords.Setting: Endoscopic biopsies of the olfactory epithelium were obtained from volunteer patients undergoing elective nasal sinus surgery. After long-term culture, as previously described, more than 150 patient-specific hONP lines were cryopreserved. In this study, the hONP lines were obtained from cryostorage and expanded as needed. Main outcome measures: Immunofluorescent characterization of hONP lines was done using cell type-specific antibodies. Open field locomotion was used to determine the degree of deficit. ELISA of isolated cord segments was used to analyze neurotrophic microenvironments. Microscopic analysis was used to examine the effect of hONP on the morphological response to SCI.Results: hONP engraftment into the contusion epicenter facilitated locomotion over nonengrafted and fibroblast engrafted controls 6 weeks following engraftment in more than 40 percent of the animals. The epicenter of hONP-engrafted animals had higher absolute axonal numbers and enriched neurotrophic micro-environments when compared with the nonengrafted controls. Conclusions: The noninvasive ability to harvest and isolate hONPs coupled with their unique regenerative capacity suggests that they are ideal candidates for an autologous cell-based strategy for treatment of human SCIs.