A Precise, Controllable in vitro Model for Diffuse Axonal Injury Through Uniaxial Stretch Injury
Regarding the determination of the biomechanical parameters in a reliable in vitro cell model for diffuse axonal injury (DAI), our study aimed to demonstrate connections between those parameters and secondary axotomy through examination of morphological alterations under a variety of traumatic condi...
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Frontiers Media S.A.
2019-10-01
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| Series: | Frontiers in Neuroscience |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fnins.2019.01063/full |
| _version_ | 1819150400030244864 |
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| author | Yu Li Yu Li Chaoxi Li Chao Gan Kai Zhao Jianbin Chen Jinning Song Ting Lei |
| author_facet | Yu Li Yu Li Chaoxi Li Chao Gan Kai Zhao Jianbin Chen Jinning Song Ting Lei |
| author_sort | Yu Li |
| collection | DOAJ |
| description | Regarding the determination of the biomechanical parameters in a reliable in vitro cell model for diffuse axonal injury (DAI), our study aimed to demonstrate connections between those parameters and secondary axotomy through examination of morphological alterations under a variety of traumatic conditions. An in vitro cell model for DAI was established in primary cultured mouse neurons by uniaxial mechanical stretching of non-myelinated axons under various traumatic conditions: strain (ε) = 5, 10, 20, and 50%; strain time (t) = 500, 100, and 20 ms; strain rate ranging between 0.1 and 25 s–1. Axonal real strains (strainaxon) were measured as 4.53 ± 0.27, 9.02 ± 0.91, 17.75 ± 1.65, and 41.8 ± 4.4%. Axonal real strain rates (SRaxon) ranged between 0.096 ± 0.0054 and 20.9 ± 2.2 s–1. Results showed there was no obvious abnormality of axons with a lower strain condition (strainaxon < 17.75 ± 1.65%) during the acute phase within 30 min after injury. In contrast, acute axonal degeneration (AAD) was observed in the axons following injury with a higher strain condition (SRaxon > 17.75 ± 1.65%). In addition, the incidence and degree of AAD were closely correlated with strain rate. Specifically, AAD occurred to all axons that were examined, when ε = 50% (strainaxon = 41.8 ± 4.4%) for 20 ms, while no spontaneous rupture was observed in those axons. Besides, the concentration of Ca2+ within the axonal process was significantly increased under such traumatic conditions. Moreover, the continuity of axon cytoskeleton was interrupted, eventually resulting in neuronal death during subacute stage following injury. In this study, we found that there is a minimum strain threshold for the occurrence of AAD in non-myelinated axons of primary cultured mouse neurons, which ranges between 9.02 ± 0.91 and 17.75 ± 1.65%. Basically, the severity of axonal secondary axotomy post DAI is strain rate dependent under a higher strain above the threshold. Hence, a reliable and reproducible in vitro cell model for DAI was established, when ε = 50% (strainaxon = 41.8 ± 4.4%) for 20 ms. |
| first_indexed | 2024-12-22T14:16:54Z |
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| id | doaj.art-fd48eb0cd2d14a1c8261d416cf643b5f |
| institution | Directory Open Access Journal |
| issn | 1662-453X |
| language | English |
| last_indexed | 2024-12-22T14:16:54Z |
| publishDate | 2019-10-01 |
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| spelling | doaj.art-fd48eb0cd2d14a1c8261d416cf643b5f2022-12-21T18:23:06ZengFrontiers Media S.A.Frontiers in Neuroscience1662-453X2019-10-011310.3389/fnins.2019.01063446348A Precise, Controllable in vitro Model for Diffuse Axonal Injury Through Uniaxial Stretch InjuryYu Li0Yu Li1Chaoxi Li2Chao Gan3Kai Zhao4Jianbin Chen5Jinning Song6Ting Lei7Department of Neurosurgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, ChinaDepartment of Neurosurgery, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an Jiaotong University, Xi’an, ChinaDepartment of Neurosurgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, ChinaDepartment of Neurosurgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, ChinaDepartment of Neurosurgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, ChinaDepartment of Neurosurgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, ChinaDepartment of Neurosurgery, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an Jiaotong University, Xi’an, ChinaDepartment of Neurosurgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, ChinaRegarding the determination of the biomechanical parameters in a reliable in vitro cell model for diffuse axonal injury (DAI), our study aimed to demonstrate connections between those parameters and secondary axotomy through examination of morphological alterations under a variety of traumatic conditions. An in vitro cell model for DAI was established in primary cultured mouse neurons by uniaxial mechanical stretching of non-myelinated axons under various traumatic conditions: strain (ε) = 5, 10, 20, and 50%; strain time (t) = 500, 100, and 20 ms; strain rate ranging between 0.1 and 25 s–1. Axonal real strains (strainaxon) were measured as 4.53 ± 0.27, 9.02 ± 0.91, 17.75 ± 1.65, and 41.8 ± 4.4%. Axonal real strain rates (SRaxon) ranged between 0.096 ± 0.0054 and 20.9 ± 2.2 s–1. Results showed there was no obvious abnormality of axons with a lower strain condition (strainaxon < 17.75 ± 1.65%) during the acute phase within 30 min after injury. In contrast, acute axonal degeneration (AAD) was observed in the axons following injury with a higher strain condition (SRaxon > 17.75 ± 1.65%). In addition, the incidence and degree of AAD were closely correlated with strain rate. Specifically, AAD occurred to all axons that were examined, when ε = 50% (strainaxon = 41.8 ± 4.4%) for 20 ms, while no spontaneous rupture was observed in those axons. Besides, the concentration of Ca2+ within the axonal process was significantly increased under such traumatic conditions. Moreover, the continuity of axon cytoskeleton was interrupted, eventually resulting in neuronal death during subacute stage following injury. In this study, we found that there is a minimum strain threshold for the occurrence of AAD in non-myelinated axons of primary cultured mouse neurons, which ranges between 9.02 ± 0.91 and 17.75 ± 1.65%. Basically, the severity of axonal secondary axotomy post DAI is strain rate dependent under a higher strain above the threshold. Hence, a reliable and reproducible in vitro cell model for DAI was established, when ε = 50% (strainaxon = 41.8 ± 4.4%) for 20 ms.https://www.frontiersin.org/article/10.3389/fnins.2019.01063/fulltraumatic brain injury (TBI)diffuse axonal injury (DAI)traumatic axonal injury (TAI)stretchin vitro model |
| spellingShingle | Yu Li Yu Li Chaoxi Li Chao Gan Kai Zhao Jianbin Chen Jinning Song Ting Lei A Precise, Controllable in vitro Model for Diffuse Axonal Injury Through Uniaxial Stretch Injury Frontiers in Neuroscience traumatic brain injury (TBI) diffuse axonal injury (DAI) traumatic axonal injury (TAI) stretch in vitro model |
| title | A Precise, Controllable in vitro Model for Diffuse Axonal Injury Through Uniaxial Stretch Injury |
| title_full | A Precise, Controllable in vitro Model for Diffuse Axonal Injury Through Uniaxial Stretch Injury |
| title_fullStr | A Precise, Controllable in vitro Model for Diffuse Axonal Injury Through Uniaxial Stretch Injury |
| title_full_unstemmed | A Precise, Controllable in vitro Model for Diffuse Axonal Injury Through Uniaxial Stretch Injury |
| title_short | A Precise, Controllable in vitro Model for Diffuse Axonal Injury Through Uniaxial Stretch Injury |
| title_sort | precise controllable in vitro model for diffuse axonal injury through uniaxial stretch injury |
| topic | traumatic brain injury (TBI) diffuse axonal injury (DAI) traumatic axonal injury (TAI) stretch in vitro model |
| url | https://www.frontiersin.org/article/10.3389/fnins.2019.01063/full |
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