Cacciamani Andrea, Parravano Mariacristina, Scarinci Fabio, Esposito Graziana, Varano Monica, Micera Alessandra
a IRCCS , G.B. Bietti Foundation , Rome , Italy.
Curr Eye Res. 2016 Jul;41(7):971-6. doi: 10.3109/02713683.2015.1080282. Epub 2015 Oct 15.
To set-up a simple technique for collecting spontaneous vitreal reflux (VR) in patients undergoing intravitreal injection. Both total protein concentration and vascular endothelial growth factor (VEGF)/Interleukin 13 (IL13) levels were used to validate the technique.
Sixty consecutive patients with neovascular age-related macular degeneration (nAMD, vitreal reflux drop, VR) and 10 patients underwent vitrectomy for macular hole (whole vitreous removal) were enrolled for the study as controls. Thirty-three out of 60 patients were also subjected to tear sampling. VR sampling was performed after the intravitreal injection. Four sampling tools (10 Schirmer strips, 10 microsponges, 20 millipore filters; 20 micropipettes) were tested. Analysis of protein concentration/composition was performed between VR samples and vitreous samples to analyze the difference. The concentration of VEGF and IL 13 levels between cases and control samples were compared.
Millipore and micropipette techniques allowed the collection of higher protein concentrations in VR samples, comparison of both protein concentrations revealed no significant difference in the protein profile. However, the micropipette sampling was found easier to perform and did not require additional protein extraction from a solid support (membrane). Indeed, tear proteins and drug contaminants were not detected in micropipette samples. Increased VEGF levels were detected in naive VR group and to a less extend in VR group of nAMD patients undergoing intravitreal injection, with respect to the controls (macular holes). No significant differences in IL13 levels were quantified in nAMD sub-groups, as compared to naive and controls.
Overall, we provide evidence for a safe method for sampling VR at the end of intravitreal injection. This procedure might represent an interesting approach either for the prognosis of disease or monitoring the efficacy of intravitreal therapy.
建立一种简单的技术，用于收集接受玻璃体内注射患者的自发性玻璃体反流（VR）。使用总蛋白浓度和血管内皮生长因子（VEGF）/白细胞介素13（IL13）水平来验证该技术。
连续纳入60例患有新生血管性年龄相关性黄斑变性（nAMD，玻璃体反流阳性，VR）的患者和10例行黄斑裂孔玻璃体切除术（全玻璃体切除）的患者作为对照。60例患者中有33例还进行了泪液采样。在玻璃体内注射后进行VR采样。测试了四种采样工具（10条泪液试纸、10个微海绵、20个微孔滤膜；20个微量移液器）。对VR样本和玻璃体样本之间的蛋白质浓度/组成进行分析，以分析差异。比较病例组和对照组样本中VEGF和IL 13水平的浓度。
微孔滤膜和微量移液器技术能够在VR样本中收集到更高的蛋白质浓度，两种蛋白质浓度的比较显示蛋白质谱无显著差异。然而，发现微量移液器采样更容易操作，并且不需要从固体支持物（膜）中额外提取蛋白质。实际上，在微量移液器样本中未检测到泪液蛋白和药物污染物。与对照组（黄斑裂孔）相比，在未治疗的VR组以及接受玻璃体内注射的nAMD患者的VR组中检测到VEGF水平升高，但升高程度较小。与未治疗组和对照组相比，nAMD亚组中IL13水平没有显著差异。
总体而言，我们提供了在玻璃体内注射结束时安全采样VR的方法的证据。该程序可能是一种用于疾病预后或监测玻璃体内治疗效果的有趣方法。
