Establishment of a mechanical injury model of rat hippocampal neurons in vitro.
OBJECTIVE
To establish a simple, reproducible, and practical mechanical injury model of hippocampal neurons of Sprague-Dawley rats in vitro.


METHODS
Hippocampal neurons isolated from 1-2-day old rats were cultured in vitro. Mild, moderate and severe mechanical injuries were delivered to the neurons by syringe needle tearing, respectively. The control neurons were treated identically with the exception of trauma. Cell damage was assessed by measuring the Propidium Iodide (PI) uptaking at different time points (0.5, 1, 6, 12 and 24 hours) after injury. The concentration of neuron specific enolase was also measured at some time points.


RESULTS
Pathological examination showed that degeneration, degradation and necrosis occurred in the injured cultured neurons. Compared with the control group, the ratio of PI-positive cells in the injured groups increased significantly after 30 minutes of injury (P<0.05). More severe the damage was, more PI-positive neurons were detected. Compared with the control group, the concentration of neuron specific enolase in the injured culture increased significantly after 1 hour of injury (P<0.05).


CONCLUSIONS
The established model of hippocampal neuron injury in vitro can be repeated easily and can simulate the damage mechanism of traumatic brain injury, which can be used in the future research of traumatic brain injury.
Yang
Xiao-feng
XF
Department of Neurosurgery, Second Affiliated Hospital, Medical College of Zhejiang University, Hangzhou 310009, China. yangxf@zju.edu.cn
Cao
Fei
F
Pan
De-sheng
DS
Liu
Wei-guo
WG
Hu
Wei-wei
WW
Zheng
Xiu-jue
XJ
Zhao
Xue-qun
XQ
Lü
Shi-ting
ST
D000704:Analysis of Variance; D000818:Animals; D001930:Brain Injuries / Q000201:enzymology / Q000473:pathology*; D004867:Equipment Design; D006624:Hippocampus / Q000201:enzymology / Q000293:injuries*; D066298:In Vitro Techniques; D009474:Neurons / Q000201:enzymology / Q000473:pathology*; D010751:Phosphopyruvate Hydratase / Q000096:biosynthesis; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results
D010751:Phosphopyruvate Hydratase