Eye Institute and Department of Ophthalmology, Eye and ENT Hospital, Fudan University, Shanghai, China.
NHC Key Laboratory of Myopia (Fudan University), Key Laboratory of Myopia, Chinese Academy of Medical Sciences, Shanghai, China.
Cornea. 2024 Oct 1;43(10):1285-1290. doi: 10.1097/ICO.0000000000003615. Epub 2024 Jul 5.
To evaluate changes of hydroxyproline concentration and its influencing factors of small incision lenticule extraction (SMILE)-derived corneal stromal lenticules with different preservation methods.
A total of 390 corneal stromal lenticules of 195 patients were derived from SMILE surgeries. Thirty of the lenticules were classified as the fresh (control) group, and the rest were randomly and evenly divided and stored in anhydrous glycerol, silicone oil, Optisol, and cryopreservation for 1 day, 1 week, or 1 month. A hydroxyproline assay kit (ab222941, Abcam) was used to measure the hydroxyproline concentration in each preservation method. Concentrations of MMP-2, TIMP-2, TNFα, TGFβ2, and reactive oxygen species were also evaluated.
In the anhydrous glycerol group, the concentration of hydroxyproline decreased within 1 week (fresh: 1 dΔ = 0.229, P < 0.001*; 1 d - 1 wΔ = 0.055, P < 0.001*) while that in the silicone oil group remained stable in 1 week (1 d - 1 wΔ = -0.005, P = 0.929) and decreased significantly in 1 m (1 m - 1 wΔ = -0.041, P = 0.003*). The sequence of hydroxyproline concentration in the Optisol group was 1 m > 1 day > 1 week. Hydroxyproline concentration in the cryopreservation group decreased within 1 m. Hydroxyproline concentration was highest in the Optisol group and lowest in the anhydrous glycerol group under the same preservation time. Hydroxyproline concentration was negatively correlated with MMP-2 (r = -0.16, P = 0.421) and TIMP-2 (r = -0.56, P = 0.002*) while MMP-2 and TNFα (r = 0.17, P = 0.242), TIMP-2 and TGFβ2 (r = 0.21, P = 0.207), and TNFα and reactive oxygen species (r = 0.52, P = 0.007*) were positively correlated.
More collagen was retained in SMILE lenticules preserved in Optisol under the same preservation time. The mechanism of the changes of collagen in preserved SMILE-derived lenticules and oxidative stress requires additional investigation.
评估不同保存方法的小切口微透镜提取（SMILE）衍生角膜基质微透镜中羟脯氨酸浓度的变化及其影响因素。
共收集 195 例患者的 390 个角膜基质微透镜，其中 30 个微透镜被归类为新鲜（对照）组，其余微透镜被随机平均分为无水甘油组、硅油组、Optisol 组和冷冻组，分别保存 1 天、1 周和 1 个月。使用羟脯氨酸测定试剂盒（ab222941，Abcam）测定每种保存方法中羟脯氨酸的浓度。还评估了 MMP-2、TIMP-2、TNFα、TGFβ2 和活性氧的浓度。
在无水甘油组中，羟脯氨酸浓度在 1 周内下降（新鲜组：1 dΔ=0.229，P<0.001*；1 d-1 wΔ=0.055，P<0.001*），而硅油组在 1 周内保持稳定（1 d-1 wΔ=-0.005，P=0.929），在 1 个月时显著下降（1 m-1 wΔ=-0.041，P=0.003*）。Optisol 组羟脯氨酸浓度的顺序为 1 m>1 d>1 w。冷冻组的羟脯氨酸浓度在 1 m 内下降。在相同的保存时间内，羟脯氨酸浓度在 Optisol 组最高，在无水甘油组最低。羟脯氨酸浓度与 MMP-2（r=-0.16，P=0.421）和 TIMP-2（r=-0.56，P=0.002*）呈负相关，而 MMP-2 和 TNFα（r=0.17，P=0.242）、TIMP-2 和 TGFβ2（r=0.21，P=0.207）以及 TNFα和活性氧（r=0.52，P=0.007*）呈正相关。
在相同的保存时间内，保存在 Optisol 中的 SMILE 微透镜中保留了更多的胶原蛋白。需要进一步研究保存的 SMILE 衍生微透镜中胶原蛋白变化和氧化应激的机制。
