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Intravitreal triamcinolone injection combined with or without macular grid laser photocoagulation to treat macular edema

Yajian DUAN ; Yading JIA ; Mianhua ZHANG

Chinese Journal of Ocular Fundus Diseases.2009;25(4):270-274. doi:10.3760/cma.j.issn.1005-1015.2009.04.07

Objective To compare the efficacy of intravitreal triamcinolone(IVTA) injection and IVTA combined with macular laser grid photocoagulation (MLGP) to treat macular edema.Methods Consecutive 89 patients (109 eyes)diagnosed with macular edema by examinations of ocular fundus and optical coherence tomography (OCT).The visual acuity was hand moving- 0.8 (0.19±0.13);the intraocular pressure(IOP)ranged from 7 mm Hg to 21 mm Hg(1 mm Hg=0.133 kPa)and the average IOP was 13.78 mm Hg.All the patients received OCT and microperimetry examinations,the central macular thickness was (570±182) μm;the average light sensitivity was (5.07±3.94) dB and the fixation percentage was 70.67% within 4 ° area around the macular fovea.All the patients received IVTA treatment,39 patients (48 eyes)further received MLGP 1 month later (IVTA-MLGP group).The remaining 50 patients (61 eyes) without MLGP treatment was the IVTA group.Best corrected visual acuity (BCVA),lOP,lens,OCT and microprimetry examinations before and after IVTA (1,3,6,12 months) were followed and analyzed.Results On the 12th months,the BCVA in IVTA-MLGP and IVTA group was (0.41±0.20),(0.24±0.19) respectively (P<0.05);the central macular thickness was (309±187) and (487±206) μm respectively(P<0.05);the mean light sensitivity of 4° central macular was (8.24±4.64)and(6.30±3.22) dB respectively (P<0.05);the fixation percentage was (87.01±19.70)% and (78.85±20.41) % respectively (P<0.05).During the follow-up recurrent macular edema was noticed in 28 eyes of IVTA group and 8 eyes of IVTA-MLGP group.Conclusions IVTA combined with MLG was more effective than IVTA to cure macular edema.

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Optic nerve injury model in rats set up by fluid percussion brain injury device

Hua YAN ; Linlin ZHANG ; Fengrong LI

Chinese Journal of Ocular Fundus Diseases.2009;25(4):301-305. doi:10.3760/cma.j.issh.1005-1015.2009.04.14

Objective To observe whether the animal model of optic nerve injury in rats can be set up by fluid percussion brain injury device (FPI) or not.Methods Seventy-one healthy female Wister rats were randomly divided into 2 groups,inlcuding model group with 66 rats and control group with 5 rats.The rats in model group were randomly divided into 3 groups.Eight rats in group 1 were examined by flash-visual evoked potential (F-VEP) and magnetic resonance imaging (MRI) examines before and 1,3 days,1,2,4,6,and 8 weeks after injury;56 rats in group 2 were randomly divided into 7 subgroups with 8 rats in each subgroup,and were detected by histopathologieal and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) apoptosis examines 1,3 days,1,2,4,6,8 weeks after injury;2 rats in group 3 were examined by electron microscopy 4 and 8 weeks aher injury.According to the degree of injury,the injured eyes were divided into 2 groups including severe injury group with the beat pressure of (699.14±60.79) kPa and mild injury group with the beat pressure of (243.18±20.26) kPa.The right and left eyes in rats in each group were in severe and mild injury group,respectively.Results One day after injury,the latency duration of F-VEP prolonged in severe injury group,wich differed much form which in the normal control group (P<0.05);the amplitude was gradually reduced during the first 2 weeks after injury and kept steady after that (P>0.05).The latency duration prolonged in mild injury group,and its difference with the normal control group was statistically significant (P<0.05);the amplitude was gradually reduced during the first 4 weeks after injury and kept steady after that (P>0.05).The abnormal high signal could be seen on optic nerve 1 day after injury,and was still obvious 8 weeks later.The results of histopathological examination showed ruptured capillary in ganglion cell layer 1 day after injury;retinal ganglion ceils without nucleus could be seen 4 weeks after injury.The apoptosis of positive cells was found in each layer of the retina 3 days after injury.TUNEL results indicated that the number of apoptotic positive cells increased significantly 1-2 weeks after injury.Conclusion An animal model of optic nerve injury can be successfully set up using FPI in rats.

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Feasibility study of differentiation of invitro induced rat bone marrow-derived mesenchymal stem cells into retinal pigment epithelial cells

Fei GAO ; Fangtian DONG

Chinese Journal of Ocular Fundus Diseases.2009;25(4):254-256. doi:10.3760/cma.j.issn.1005-1015.2009.04.03

Objective To investigate the feasibility of differentiation of invitro induced rat bone marrow-derived mesenchymal stem cells(rMSCs) into retinal pigment epithelial (RPE) cells.Methods The rMSCs from Brwon-Norway (BN) rats were isolated and cultured by adherent screening method.RPE cells lysate made by repeated freeze-thawing was put into the rMSCs culture system to identify whether the induced cells could express characteristic label cytokeratin(CK)and S-100 simultaneously or not.Results The growth rate of rMSCs induced by RPE cells lysate was slower and protuberant burr surrounded the fusiform cells.The results of immunoblotting and double immunofluorescence showed that partial induced cells expressed CK and S-100 simultaneously.The result of flow cytometry indicated that 14.1% induced cells expressed CK and S-100 simultaneously.Conclusion Induced by RPE cells lysate,rMSCs can differentiate into RPE cells.

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Somatic reprogramming:new breakthrough in researches on stem cells

Jian GE

Chinese Journal of Ocular Fundus Diseases.2009;25(4):247-248. doi:10.3760/cma.j.issn.1005-1015.2009.04.01

Replacement therapy of stem cells transplantation represents a potential treatment for neural retinal diseases.Despite the encouraging results in laboratory,the clinical application of cells replacement therapy is still difficult because the limitation of seed cells,immunologic rejection,oncogenicity and ethical problems,etc.Recent breakthrough in somatic reprogramming provides a promising solution overcoming these obstacles.Further researches on virus-free reprogramming will make the clinical application of stem cell replacement therapy possible.

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Transplantation of bone-marrow derived mesenchymal stem cells to treat rat retinal degeneration

Yuan GUAN ; Lu CUI ; Yalan WU ; Weiya LI ; Guotong XU

Chinese Journal of Ocular Fundus Diseases.2009;25(4):249-253. doi:10.3760/cma.j.issn.1005-1015.2009.04.02

Objective To observe the effects of subretinal transplantation of rat mesenchymal stem cells (rMSCs) on Sodium Iodate (SI)-induced retinal degeneration.Methods One hundred and twenty Brown-Norway (BN) rats were divided into three groups including SI injection group,rMSCs transplantation group and normal control group,each with 40 rats.The retinal degeneration was induced by caudal vein injection of SI.The retinal pigment epithelium(RPE)and neural retinal were evaluated by ocular fundus photograph,fluorescein fundus angiography (FFA),electroretinogram (ERG) and histological approach,and TUNEL(terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling).CM-Dilprelabeled primary rMSCs were transplanted into the subretinal space of Sl-induced rats.The survival,integration,and differentiation of rMSCs were observed between 14 day to 60 day after the transplantation.Results The rat retinal function was gradually reduced afterl4 days of SI injection,with a time-dependent manner.After the RPE cells were damaged,the outer segments of photoreceptors became disrupted and shortened until karyopyknosis.The nuclear morphology and positive TUNEL labeling indicated that the death of photoreceptor cells was apoptosis.After rMSCs transplantation,CM-DiI labeled donor cells were observed to be scattered in the subretinal space and expressed RPE cell markers.Average amplitude of bwave and Ops (oscillation potential) in ERG improved 27.80%,59.38% respectively after rMSCs transplantation.Conclusions Transplanted rMSCs can survive in subretinal space and differentiate into RPE cells,thus cure SI- induced retinal degeneration.

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Apoptosis of retinal capillary cells and expression of p53 in rats with spontaneous hypertension after ischemic reperfusion injury

Saibin WANG ; Bin JI ; Bixin CHEN ; Mingying ZHANG ; Xiaoyan HUANG ; Deye YANG

Chinese Journal of Ocular Fundus Diseases.2009;25(3):198-201. doi:10.3760/cma.j.issn.1005-1015.2009.03.011

Objective To investigate the effect of apoptosis-related gene p53 on the apoptosis of retinal capillary cells in rats with spontaneously hypertensive (SHR) after ischemic reperfusion injury. Methods A total of 60 SHR rats were randomly divided into sham group (SHR-SH) and retinal ischemie reperfusion group (SHR-RIR), which were subdivided into 5 subgroups according to the time after RIR: 2, 6, 24, and 72 hours and 7 days, with 6 rats in each subgroup. Another 60 Wistar-Kyoto (WKY) rats were divided into the same groups as the SHR rats as the control. The RIR model was set up. The apoptosis of retinal capillary cells was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) methods and the expression of p53 was determined by streptavidin-perosidase (SP) immunohistochemistry. Results The apoptosis rate of retinal capillary cells in the 5 SHR-RIR groups was (8.64±0.56)%, (14.92±0.99)%, (24.72±2.98)%, (16.53±1.80)%, and (7.12±1.10)%,respectively. The expression of p53 in SHR-RIR groups increased at the 2nd hour after RIR, reached the peak at the 24th hour, kept the high level at the 72nd hour, and remained a little at the 7th day, which was significantly different from which in the SHR-SH groups (P<0. 01). The expression of p53 were higher in SHR-IR groups than that in the WKY-RIR groups (P<0. 01). Conclusions p53 may play a part in RIR injury by inducing or promoting apoptosis. The apoptosis of retinal capillary cells after RIR is more severe under the hypertension, and reaches the peak at the 24 hour after RIR.

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The role of receptor protein tyrosine kinase MERTK and intracellular Ca2+ playing in the phagocytosis of human retinal pigment epithelial cells

Yuzhao SUN ; Jing HONG ; Gang AN

Chinese Journal of Ocular Fundus Diseases.2009;25(3):193-197. doi:10.3760/cma.j.issn.1005-1015.2009.03.010

Objective To investigate the role of intraeellular Ca2+ and MERTK in the phagocytosis of human retinal pigment epithelial (RPE) cells, and reveal the relationship between MERTK and intraeellular Ca2+. Methods The euhured RPE cells were incubated with rod outer segments (ROS) at 37℃, the phagoeytosis was terminated at different incubation time points. The concentration of intraeellular Ca2+ was assayed by Fluro-3/AM loading methods combined with fluorescence microscope and CCD system, and the mRNA level of MERTK gene was measured by reverse transcription polymerase chain reaction (RT-PCR). Treating the RPE cells with stimulator (A23187)or inhibitor(verapamile)of intraeellular Ca2+ to observe the changes of MERTK gene expression. Results ROS adhered to hRPE cells at the 15th minute, and the ingestion saturated at the 24th hour. The concentration of intracellular Ca2+ increased at the 15th minute, and kept the high level in 24 hours. The level of MERTK mRNA increased at the 5th minute, and kept the high level duration the whole incubation. When RPE cells were treated by A23187, the expression of MERTK increased in a dose-dependent manner. After RPE cells was pretreated by A23187, the expression level of MERTK was higher in the proceeding incubation groups than which in the control group except at the 3rd hour. When RPE cells were treated by verapamil, the expression level of MERTK decreased in a dose-dependent manner. After RPE cells were pretreated by verapamil , the expression level of MERTK was lower in all the proceeding incubation groups than which in the control group (P<0.05). Conclusion MERTK gene and Ca2+ play an important role in sustaining RPE cells phagoeytizing ROS. As an up-stream regulator, the receptor tyrosine kinase MERTK keeps RPE cells phagocytizing ROS by starting the intracellular Ca2+.

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Correlation between the central macular thickness and the visual function in patients with macular edema

Wenqiu WANG ; Hong WANG ; Lei ZHANG ; Yuan JIANG ; Jin QIAN ; Weijun WANG ; Fenghua WANG ; Ying WU ; Xiaodong SUN

Chinese Journal of Ocular Fundus Diseases.2009;25(3):182-184. doi:10.3760/cma.j.issn.1005-1015.2009.03.007

Objective To determine the correlation between central macular thickness (CMT) and the visual function in patients iwht macular edema (ME). Methods The clinical data of 42 eyes of 40 patients with ME which were examined by optical coherence tomography (OCT) and microperimetry (MP-1) wereretrospectively analyzed. In 40 patients (42 eyes), diabetic ME (DME) was in 27 eyes,branch retinal vein occlusion was in 11eyes, and central retinal vein occlusion was in 4 eyes. All of the eyes had undergone OCT,MP-1 and best-corrected visual acuity (BCVA) test. Central macular thickness (CMT) was measured by fast macular scans using OCT. Retinal sensitivity (MS) and fixation patterns were evaluated by Mp-1.The position was chosen :2 disc diameters (DD) temporal to the disc and one third of a DD inferior to the centre of the disc. Results The correlation between CMT and BCVA is not significant (r=-0. 429, P=0. 069) as well as the correlation between CMT and MS (r=-0. 433,P=0. 058). The difference of CMT between the unstable and stable group was significant (F = 3. 262, P = 0. 039). The difference of CMT between the central fixation group and preferred retinal locus (PRL) group was significant (F=3. 173,P=0. 044). Conclusions BCVA and MS have no significant correlation with CMT. When CMT increases, the fixation stability decreases, fixation location, changes, and PRL occurs.

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Optical coherence tomography: a new era of ocular-fundus imageological technology

Xiaoxin LI ; Weizhi CHEN

Chinese Journal of Ocular Fundus Diseases.2009;25(3):161-163. doi:10.3760/cma.j.issn.1005-1015.2009.03.001

Optical coherence tomography (OCT) has developed from time-domain into Fourier-domain OCT (FD-OCT) which indicates clearer details and higher resolution of images. FD-OCT can indicate the structure and pathological changes of each retinal layer, and reveal the retinal external limiting membranes and changes of inner-and outer-segment of visual cells by 3D solid reconstruction. FD-OCT not only provide detailed information of the images for the clinical diagnosis, but also help us investigting the characteristics and pthological mechanisms of ocular fundus diseases, which lead us to a new era of technology of observation on ocualr fundus diseases. In the application, we should pay attention to the significance of different colors of OCT images, and focus on the cohenrence of the position in the image acquistion during the follow-up period. Dynamic observation on the lesions by FD-OCT and aggregated anaylsis of resutls of several imageological examination would be the development direction of imageological examination of ocular fundus diseases.

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Insulin has no effect on vascular endothelial growth factor expression in the retina of streptozotocin-induced diabetic rats

Haixiang WU ; Xin XIA ; Qing GU ; Xun XU

Chinese Journal of Ocular Fundus Diseases.2009;25(3):202-205. doi:10.3760/cma.j.issn.1005-1015.2009.03.012

Objective To investigate if insulin can affect the expression of vascular endothelial growth factor (VEGF) in the retina of streptozotocin-induced diabetic rats. Methods A total of 60 male Sprague-Dawley rats were randomly divided into sodium citrate buffer control group (CIT-CON, n= 30) and STZ-induced diabetic group (STZ-DM, n=30). At the 16th week, 24 rats from CIT-CON group at random were randomly divided to group A (sodium citrate buffer control group, n = 12) and group B (sodium citrate buffer plus insulin group, n= 12). The remaining 6 rats from as CIT-CON group served as negative control. At the same time, 24 rats from STZ-DM group at random were randomly divided to group C (STZ-induced diabetic group, n= 12) and group D (STZ-induced diabetic plus insulin group, n= 12). The remaining 6 rats from STZ-DM group also served as negative control. 4 IU of insulin was injected subcutaneously to rats of group B and D. Immunohistochemistry, Western blot and Real-time polymerase chain reaction (RT-PCR) were used to measure the expression level of VEGF protein and mRNA respectively. RESULTS Insulin significantly increased the VEGF mRNA (7.71 ± 0.25 vs 5.36 ±0. 37, t test P< 0. 05) and protein expression (0. 4925 ± 0. 0122 vs 0. 4272 ± 0. 0110, t test P< 0. 05) in the retina of CIT-CON rats.However, in retina of STZ-DM rats, insulin had no effect on VEGF mRNA (8. 92±0. 27 vs 9. 05±0. 28, t test, P>0. 05) and protein expression (0. 5152±0. 0109 vs 0. 5099±0. 0100, t test P>0.05). Conclusions Insulin had no effect on VEGF expression in the retina of STZ-DM rats.

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