ABSTRACT
BACKGROUND@#Repair of long-distance peripheral nerve defects remains an important clinical problem. Nerve grafts incorporated with extracellular vesicles (EVs) from various cell types have been developed to bridge peripheral nerve defects. In our previous research, EVs obtained from skin-derived precursor Schwann cells (SKP-SC-EVs) were demonstrated to promote neurite outgrowth in cultured cells and facilitate nerve regeneration in animal studies. @*METHODS@#To further assess the functions of SKP-SC-EVs in nerve repair, we incorporated SKP-SC-EVs and Matrigel into chitosan nerve conduits (EV-NG) for repairing a 15-mm long-distance sciatic nerve defect in a rat model. Behavioral analysis, electrophysiological recording, histological investigation, molecular analysis, and morphometric assessment were carried out. @*RESULTS@#The results revealed EV-NG significantly improved motor and sensory function recovery compared with nerve conduits (NG) without EVs incorporation. The outgrowth and myelination of regenerated axons were improved, while the atrophy of target muscles induced by denervation was alleviated after EVs addition. @*CONCLUSION@#Our data indicated SKP-SC-EVs incorporation into nerve grafts represents a promising method for extended peripheral nerve damage repair.
ABSTRACT
AIM: To investigate the antitumor effect and mechanism of trichosanthin (TCS) on melanoma B-16 cells. METHODS: (1) The injury of B-16 cells by trichosanthin was observed with SCGE and hoechst33258 staining method. (2) LCSM and specificity fluorescent probe Fluo-3/AM, H2DCF-DA, DAF-FM diacetate were applied to analyze the dynamic changes of Ca2+, ROS and NO in single cell cultured with TCS. Simultaneously, the relationship between ROS, NO and increase of Ca2+ was also revealed. RESULTS: (1)When treated with TCS (50 mg/L) for 3 h and 6 h, neither cytotoxicity assay nor SCGE showed the differences compared with control group. After 12 h incubation, specificity phenomena of DNA injury-comet tail appeared in SCGE and chromatin condensation even apoptotic body formation were seen by Hoechst33258 staining. (2) TCS (50 mg/L) evoked rapid enhancement of the production of Ca2+, ROS and NO in the cell and the differences between TCS and control group had statistical significance (P