Centre of Ophthalmology, University Eye Hospital Tübingen, 72076, Tübingen, Germany.
Graefes Arch Clin Exp Ophthalmol. 2013 Jun;251(6):1613-9. doi: 10.1007/s00417-012-2255-6. Epub 2013 Jan 11.
Several iatrogenic risk factors during pars plana vitrectomy (PPV) could cause damage to the retina. One mechanism is excitotoxicity. Therefore, neuroprotective irrigation solutions would be desirable.
Retinal ganglion cells (RGC-5) and retinal whole mounts were incubated in standard irrigation solution (SIS) and Dulbecco's Modified Eagle Medium (DMEM). Cell viability, cell amount, cell survival and caspase 3/7 activity were measured by MTS-Test, crystal-violet staining, Annexin-V/PI flow cytometry and caspase 3/7 activity assay, respectively. The morphology and the function of retinal whole mounts were analysed by Live/Dead(TM) staining and by the b-wave and a-wave of the electroretinogram (ERG).
Under excitotoxic conditions (10 mM and 12 mM glutamate) RGC-5 cells incubated in SIS showed a statistically significant reduction in cell viability, cell amount, cell survival and caspase 3/7 activity compared to DMEM. Furthermore, the incubation of retinal whole mounts in DMEM resulted in a significant decrease of cell death under excitotoxic (250 μM glutamate) and standard conditions compared to SIS. ERG b-wave recordings revealed good functional preservation of retinal whole mounts in DMEM, but loss in SIS.
DMEM seems to support retinal cells very well and to be strongly protective against excitotoxicity. Therefore, DMEM may be considered as possible neuroprotective irrigation solution for PPV.
在平坦部玻璃体切除术（PPV）过程中，有几个医源性危险因素可能会对视网膜造成损害。其中一种机制是兴奋性毒性。因此，需要使用具有神经保护作用的灌洗液。
将视网膜神经节细胞（RGC-5）和视网膜全铺片分别孵育于标准灌洗液（SIS）和 Dulbecco 改良 Eagle 培养基（DMEM）中。通过 MTS 试验、结晶紫染色、Annexin-V/PI 流式细胞术和 caspase 3/7 活性测定分别测量细胞活力、细胞数量、细胞存活率和 caspase 3/7 活性。通过 Live/Dead(TM)染色和视网膜电图（ERG）的 b 波和 a 波分析视网膜全铺片的形态和功能。
在兴奋性毒性条件下（10mM 和 12mM 谷氨酸），与 DMEM 相比，孵育于 SIS 中的 RGC-5 细胞的细胞活力、细胞数量、细胞存活率和 caspase 3/7 活性均显著降低。此外，与 SIS 相比，DMEM 孵育的视网膜全铺片在兴奋性毒性（250μM 谷氨酸）和标准条件下细胞死亡明显减少。ERG b 波记录显示，DMEM 可很好地维持视网膜全铺片的功能，但 SIS 则会导致功能丧失。
DMEM 似乎能很好地支持视网膜细胞，并且对兴奋性毒性有很强的保护作用。因此，DMEM 可被视为 PPV 中具有神经保护作用的潜在灌洗液。
