Nerve conduits have been a viable alternative to the ‘gold standard’ autograft for treating small peripheral nerve gap injuries. However, they often produce inadequate functional recovery outcomes and are ineffectiv...Nerve conduits have been a viable alternative to the ‘gold standard’ autograft for treating small peripheral nerve gap injuries. However, they often produce inadequate functional recovery outcomes and are ineffective in large gap injuries. Ridge/groove surface micropatterning has been shown to promote neural cell orientation and guide growth. However, optimization of the ratio of ridge/groove parameters to promote orientation and extension for dorsal root ganglion （DRG） cells on poly（lactic-co-glycolic acid） （PLGA） films has not been previously conducted. Photolithography and micro-molding were used to define various combinations of ridge/groove dimensions on PLGA films. The DRG cells obtained from chicken embryos were cultured on micropatterned PLGA films for cell orientation and migration evaluation.Biodegradation of the films occurred during the test period, however, this did not cause deformation or distortion of the micropatterns. Results from the DRG cell orientation test suggest that when the ridge/groove ratio equals 1 （ridge/groove width parameters are equal, i.e., 10 μm/10 μm （even））, the degree of alignment depends on the size of the ridges and grooves, when the ratio is smaller than 1 （groove controlled） the alignment increases as the ridge size decreases, and when the ratio is larger than 1 （ridge controlled）, the alignment is reduced as the width of the grooves decreases. The migration rate and neurite extension of DRG neurons were greatest on 10 μm/10 μm and 30 μm/30 μm micropatterned PLGA films. Based on the data, the 10 μm/10 μm and 30 μm/30 μm micropatterned PLGA films are the optimized ridge/groove surface patterns for the construction of nerve repair devices.展开更多
以0、3.7×103、3.7×104、3.7×105及3.7×106Bq/mL的氚水(HTO)照射原代培养的神经细胞,照后24小时用Leica AF 6000活细胞工作站测细胞迁移距离,Fluo-3-AM荧光探针负载和流式细胞仪测定细胞内游离钙离子浓度,Western b...以0、3.7×103、3.7×104、3.7×105及3.7×106Bq/mL的氚水(HTO)照射原代培养的神经细胞,照后24小时用Leica AF 6000活细胞工作站测细胞迁移距离,Fluo-3-AM荧光探针负载和流式细胞仪测定细胞内游离钙离子浓度,Western blot法分析细胞内β-tubulin和NCAM蛋白变化,以探讨氚β辐射影响神经细胞迁移的机制。结果发现,随着氚水浓度增大,氚照射剂量的增加,神经细胞迁移距离逐渐减小,细胞内游离钙离子逐渐增多,β-tubulin和NCAM表达逐渐减弱。提示氚β辐射可使神经细胞迁移相关调控因子产生变化,影响其迁移。展开更多
文摘Nerve conduits have been a viable alternative to the ‘gold standard’ autograft for treating small peripheral nerve gap injuries. However, they often produce inadequate functional recovery outcomes and are ineffective in large gap injuries. Ridge/groove surface micropatterning has been shown to promote neural cell orientation and guide growth. However, optimization of the ratio of ridge/groove parameters to promote orientation and extension for dorsal root ganglion （DRG） cells on poly（lactic-co-glycolic acid） （PLGA） films has not been previously conducted. Photolithography and micro-molding were used to define various combinations of ridge/groove dimensions on PLGA films. The DRG cells obtained from chicken embryos were cultured on micropatterned PLGA films for cell orientation and migration evaluation.Biodegradation of the films occurred during the test period, however, this did not cause deformation or distortion of the micropatterns. Results from the DRG cell orientation test suggest that when the ridge/groove ratio equals 1 （ridge/groove width parameters are equal, i.e., 10 μm/10 μm （even））, the degree of alignment depends on the size of the ridges and grooves, when the ratio is smaller than 1 （groove controlled） the alignment increases as the ridge size decreases, and when the ratio is larger than 1 （ridge controlled）, the alignment is reduced as the width of the grooves decreases. The migration rate and neurite extension of DRG neurons were greatest on 10 μm/10 μm and 30 μm/30 μm micropatterned PLGA films. Based on the data, the 10 μm/10 μm and 30 μm/30 μm micropatterned PLGA films are the optimized ridge/groove surface patterns for the construction of nerve repair devices.
文摘以0、3.7×103、3.7×104、3.7×105及3.7×106Bq/mL的氚水(HTO)照射原代培养的神经细胞,照后24小时用Leica AF 6000活细胞工作站测细胞迁移距离,Fluo-3-AM荧光探针负载和流式细胞仪测定细胞内游离钙离子浓度,Western blot法分析细胞内β-tubulin和NCAM蛋白变化,以探讨氚β辐射影响神经细胞迁移的机制。结果发现,随着氚水浓度增大,氚照射剂量的增加,神经细胞迁移距离逐渐减小,细胞内游离钙离子逐渐增多,β-tubulin和NCAM表达逐渐减弱。提示氚β辐射可使神经细胞迁移相关调控因子产生变化,影响其迁移。
