玻璃体腔注射塞来昔布一聚乳酸羟基醋酸共聚物缓释微球的安全性研究

The effects of celecoxib-poly lactide-co-glycolide microparticles on rat retina after intravitreai injection

目的观察玻璃体腔注射塞来昔布一聚乳酸羟基醋酸共聚物缓释微球（CEL—PLGA—MS）对视网膜的安全性。方法32只雄性BrownNorway大鼠随机分为CEL—PLGA—MS组和塞来昔布（celecoxib）组，每组各16只。CEL—PLGAMS组分为4个剂量组，每组各4只。分别玻璃体腔注入含celecoxib40、80、160、320p．mol／L的聚酸乳羟基醛酸共聚物（PLGA）微球。Celecoxib组分为4个剂量组，每组各4只。分别玻璃体腔注入celecoxib40、80、160、320／~mol／L；磷酸盐缓冲液（PBS）对照组4只，2只双眼玻璃体腔注入0．01mol／LPBS溶液，正常对照组2只，双眼不处理。光相干断层扫描（0CT）测量各组视网膜厚度，光学显微镜观察各组视网膜微结构变化，透射电子显微镜观察各组视网膜细胞的超微结构变化。结果OCT检查结果显示，正常对照组和PBS对照组视网膜厚度比较，差异无统计学意义（F=0．12，P〉0．05）。注药后1周，CEL-PLGAMS、celecoxib各组视网膜厚度均大于正常对照组、PBS对照组。组问视网膜厚度比较，差异有统计学意义（F=9．62、46．13；P〈0．01）。CEL—PLGA-MS各组视网膜厚度小于celecoxib各组。组间视网膜厚度比较，差异有统计学意义（F=165．15，P〈0．01）。CEL—PLGA—MS40、80、320μmol／L组视网膜厚度大致相同。组间视网膜厚度比较，差异有统计学意义（F=4．79，P〈0．01）。Celecoxib160、320μmol／L组视网膜厚度大于celecoxib40、80umol／I。组。组间视网膜厚度比较，差异有统计学意义（F=28．10，P％0．01）。注药后2周，CEL—PLGA—Ms各组视网膜厚度与eelecoxib各组比较，差异无统计学意义（F=3．79，P〉0．05）。且较注药后1周时视网膜厚度逐渐减小，差异有统计学意义（F=7．28、103．99；P〈0．01）。注药后4周，CEL—PLGA—MS各组视网膜厚度小于celecoxib各组。组间视网膜厚度比较，差异有统计学意义（F=19．11，P％0．01）。CEL—PLGA—MS各组视网膜厚度和正常对照组、PBS对照组大致相等。组间视网膜厚度比较，差异有统计学意义（F=2．02，P〉0．05）。Celecoxib各组视网膜厚度大于正常对照组和PBS对照组。组间视网膜厚度比较，差异有统计学意义（F=3．71，P〈0．05）。光学显微镜下各组视网膜结构未见明显变化，其视网膜厚度变化与注药后第1周OCT检查结果一致。透射电子显微镜下celecoxib160、320μmol／L组视网膜毒性反应重，视网膜内层细胞超微结构比外层变化明显。CEL—PLGA-MS320μmol／L组内网状层微丝排列紊乱、微管扩张、线粒体少许空泡化。Celecoxib40、80μmol／L组和CEL—PLGA—MS40、80、160μmol／L组细胞超微结构变化轻微。结论玻璃体腔注射CEL—PLGA—MS对视网膜安全性优于玻璃体腔注射celecoxib。

Objective To investigate the effects of （CEL-PLGA MS） on rat retina after intravitreal injection. celecoxib-poly lactide-co-glycolide microparticles Methods A total of 32 male Brown Norway rats were randomly divided into CEL-PLGA-MS group and celecoxib group, 16 rats in each group. The rats in CEL-PLGA-MS group were divided into four dosage group, four rats in each group, which received intravitreal injection of PLGA with celecoxib at the concentration of 40, 80, 160, 320μmol/L, respectively. The rats in celecoxib group were divided into four dosage group, four rats in each group, which receivedintravitreal injection of celecoxib at the concentration of 40, 80, 160, 320 μmol/L, respectively. Phosphate buffer solution （PBS） was injected in two rats as PBS control group. Two rats as normal control group received no treatment. The difference of retinal thickness among groups was measured by optical coherence tomography （OCT）. The morphological and histological change of retina was evaluated under light microscope and transmission electron microscope. Results There was no difference of retinal thickness between normal control group and PBS control group （F = 0.12, P~〉 0.05）. At the first week after injection, the retinal thickness of CEL-PLGA-MS group and celecoxib group were thicker than that in normal control group and PBS control group （F = 9.62, 46.13; P〈 0.01）. The retinal thickness of celecoxib group was thicker than that in CEL-PLGA-MS group （F = 165.15, P〈 0.01）. The retinal thickness was estimated equal among 40, 80, 320 μmol/L dosage groups in CEL-PLGA MS group （F= 4.79, P^0.01）. The retinal thickness of 160, 320 μmol/L dosage group were thicker than that in 40, 80 μmol/L dosage group in celecoxib group （F=28.10, P〉0.01）. At the second week after injection, there was no difference of retinal thickness between CEL-PLGA-MS and celecoxib group （F= 3.79, P〈0. 05） ; the retinal thickness of CEL-PLGA-MS and celecoxib group became thinner gradually compare to the first week after injection （F=7.28, 103.99; P〈0.01）. At the fourth week after injection, the retinal thickness of celecoxib group was thicker than that in CEL-PLGA-MS group （F= 19. 11, P〈0. 01）. The retinal thickness of CEL-PLGA-MS group was approximately the same to normal control group and PBS control group （F=2.02, P〉0.05）. The retinal thickness of celecoxib group was thicker than that in normal control group and PBS control group. No considerable abnormality of the retina was seen by light microscope and the retinal thickness corresponded with the values measured by OCT at the first week after injection. The abnormal structures of the retina were seen in 160, 320 μmol/L dosage group of celecoxib group and inner changed evidently by the transmission electron microscope. Disordered arrangement of microfilaments, dilated microtubule and some mitoehondria vacuolation were observed in 320 μmol/L dosage group of celecoxib group. Others changed slightly. Conclusions CEL-PLGA-MS has less toxicity on the retina than free-celecoxib after intravitreal injection. The safety of intravitreal injection with CEL-PLGA-MS is better than celecoxib.