Electrical and Neurotrophin Enhancement of Neurite Outgrowth Within a 3D Collagen Scaffold
Electrical and Neurotrophin Enhancement of Neurite Outgrowth Within a 3D Collagen Scaffold
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DOI：
10.1007/s10439-014-1001-0
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发表时间：
2014-06-01
影响因子：
3.8
通讯作者：
Harkins, Amy B.
中科院分区：
文献类型：
--
作者：
Adams, Robert D.;Rendell, Sara R.;Harkins, Amy B.
电刺激和化学刺激被认为是促进神经再生和伤口愈合的有效机制。然而，目前还不清楚电刺激如何影响神经生长，特别是在神经营养因子存在的情况下。本研究的目的是探讨(1)补充脑源性神经营养因子(BDNF)和神经生长因子(NGF)对3D支架中神经突起生长的支持作用；(2)在补充神经营养素之前，短暂的低电压电刺激(ES)对神经突起生长的影响。将分离的E11鸡背根神经节(DRG)接种于1.5 mg/mLI型胶原支架内。对于神经营养因子治疗，支架在含有NGF(10 ng/mL)和/或BDNF(200 ng/mL)的培养液中孵育24小时。ES组以8~10V/m DC刺激含神经元支架10min，再与神经营养素孵育24 h。对固定的和标记的神经元进行成像，以测量轴突生长和方向性。补充BDNF在支持DRG突起生长方面不如NGF有效。与单独补充NGF相比，补充NGF之前的ES改善了DRG轴突的生长。与单独应用NGF或NGF或NGF相比，这种简单的ES联合NGF治疗是最有效的治疗方法。Brief ES对3D支架中轴突的方向性无影响。这些结果表明，ES在神经营养因子存在的情况下促进了神经突起的生长，当与神经营养因子联合治疗时，可以提供一种潜在的治疗方法来促进神经再生。
Electrical and chemical stimulation have been studied as potent mechanisms of enhancing nerve regeneration and wound healing. However, it remains unclear how electrical stimuli affect nerve growth, particularly in the presence of neurotrophic factors. The objective of this study was to explore (1) the effect of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) supplementation to support neurite outgrowth in a 3D scaffold, and (2) the effect of brief, low voltage, electrical stimulation (ES) on neurite outgrowth prior to neurotrophin supplementation. Dissociated E11 chick dorsal root ganglia (DRG) were seeded within a 1.5 mg/mL type-I collagen scaffold. For neurotrophin treatments, scaffolds were incubated for 24 h in culture media containing NGF (10 ng/mL) or BDNF (200 ng/mL), or both. For ES groups, scaffolds containing neurons were stimulated for 10 min at 8-10 V/m DC, then incubated for 24 h with neurotrophin. Fixed and labeled neurons were imaged to measure neurite growth and directionality. BDNF supplementation was not as effective as NGF at supporting DRG neurite outgrowth. ES prior to NGF supplementation improved DRG neurite outgrowth compared to NGF alone. This combination of brief ES with NGF treatment was the most effective treatment compared to NGF or BDNF alone. Brief ES had no impact on neurite directionality in the 3D scaffolds. These results demonstrate that ES improves neurite outgrowth in the presence of neurotrophins, and could provide a potential therapeutic approach to improve nerve regeneration when coupled with neurotrophin treatment.