This experiment was designed to observe the proliferative effects of platelet derived wound healing factor (PDWHF) of different concentrations on fibroblasts from rat wounds and on epithelial from human wounds. Cultured fibroblasts from rat wound and epithelia from human wound were randomly divided into three groups. (1) In blank control, the cells were treated with basic medium (BM, contains 1640/0.5% FCS); (2) the positive control, the cells were treated with 1640/10% FCS and (3) in the PDWHF group, the cells were treated respectively with BM/1% PDWHF, BM/3% PDWHF, BM/5% PDWHF, BM/7% PDWHF, BM/10% PDWHF, BM/12% PDWHF, respectively. The Cells were collected after 48 hours culturing with BM or PDWHF, and the cell proliferation was measured by MTT method according to the OD values. The result showed that the PDWHF could remarkably enhance the proliferation of fibroblasts and epithelial cells when its concentration was between 1% and 7%, which was obviously higher than that of the blank control (P lt; 0.01). When the concentration of PDWHF reached 10%, its proliferative effect was not remarkable when compared with the blank control, When the concentration of PDWHF reached 12%, it showed inhibitory effect on fibroblasts and manifested no obvious inhibitory effect on epithelial cells. It was concluded that the PDWHF was a combination of a variety of growth factors. In a certain range of concentration, the PDWHF might effectively promote the proliferation of fibroblasts and epithelial cells. Howerve, when its concentration reached to relatively higher level, its effect was not remarkable any more, or even showed inhibitory effect on cell proliferation.
Objective To investigate the feasibility of differentiation of invitro induced rat bone marrowderived mesenchymal stem cells（rMSCs） into retinal pigment epithelial （RPE） cells.Methods The rMSCs from BrwonNorway (BN) rats were isolated and cultured by adherent screening method. RPE cells lysate made by repeated freezethawing 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.
Objective To evaluate the effects of inflammatory cytokines, including tumor necrosis factorTNF-alpha; and interleukins (IL-6 and IL-8), to the expression of pigment epithelium-derived factor (PEDF) in human retinal pigment epithelium (RPE)cells. Method Cultured primary human RPE cells were treated with 20，2，0.2 , and 0.02 ng/ml of TNF-alpha;, IL-6 and IL-8 separately. The levels of PEDF expression were determined by Western blot of the supernant after 6，12，24 and 48 hours of culture. Results PEDF secretion of RPE cells was inhibited by TNF-alpha;, IL-6 and IL-8 in a time- and dose-dependent fashion. Compared with the controls, the expression of PEDF decreased significantly in 0.02 ng/ml and 6 hours group (F＝7.14, P＜0.05）, 2.00 ng/ml and 48 hours group（F=14.05，P＜0.01) , and 20.00 ng/ml and 24 hours group（F=11.53，P＜0.01). TNF-alpha; was the most strength inhibitor (F＝14,P＜0.01）.Conclusion TNF-alpha;, IL-6, and IL-8 could suppress the expression of PEDF in the cultured human RPE cells.
Objective To investigate the effect of subretinal fluid (SRF) with different grades of proliferative vitreoretinopathy (PVR) on bcl-2 oncoprotein expression in retinal pigment epithelium (RPE) cells and fib roblast (FB)． Methods Using immunohistological staining technique and Western-blotting method to detect the expression of bcl-2 protein in RPE cells and FB under the stimulation of SRF． Results The expression levels of bcl-2 increased in both types of cells to a certain extent compared with those of the control group 4 hours after the cells subjected to SRF； 36 hours later, the expression levels of bcl-2 of experimental groups decreased more quickly than those of the control group， and the control group showed relatively higher bcl-2 protein levels at the end of observation． Conclusions SRF may stimulate the e xpression of bcl-2 in RPE cells and FB under culture at early stage, but accel arate the declining of bcl-2 protein levels a certain time after subjected to SRF． (Chin J Ocul Fundus Dis, 2001,17:58-60)
Objective To investigate the spatial and temporal regulation effect of VEGF on human fetal retinal vascularization and angiogenesis. Methods The posterior segmental retinas from 54 human fetuses of the 9th week to the 40th week were studied by immunohistodhemistry standing for the expressions of VEGF and PCNA. Results 1. The distribution of VEGF espression was spiking and the peaks were during the 9th-13th and around the 26th week. 2. PCNA immunoreactivity was localized in spindle cells and vascular endothelial cells. The expression level was fluctuated during the developmental process. The peaks were during the 9th-13th and around the 21st week. In these periods, the spindle cells kept proliferating and differentiating, and remodelled subsequently to form the inner side retinal vessels. From the 26th or 34th week, the PCNA immununoreactivity is fully expressed in the vascular endothelial cells of the inner and outer margin of inner nuclear layer(INL) and kept to full terms. 3. Significant positive correlation were shown between the content of VEGF in the retina and that of PCNA in spindle cells and vascular endothelial cells(r=0.736,p＜0.01). Conclusion VEGF was positively involved in modulating human fetal retinal vascularization and angiogenesis. (Chin J Ocul Fundus Dis,1999,15:12-15)
Objective To analyze the risk factors for persistent corneal epithelial defects (PCED) after pars plana vitrectomy (PPV) in patients with proliferative diabetic retinopathy (PDR). Methods A total of 201 PDR patients (201 eyes) who received PPV were enrolled in this retrospective study. There were 86 males (86 eyes) and 115 females (115 eyes). The patients aged from 30 to 81 years, with the mean age of (57.94±9.65) years. Among them, 159 patients were ≥50 years of age, and 42 patients were ＜50 years of age. There were 36 patients with HbA1c ＜7.0%, 165 patients with HbA1c ≥7.0%. There were 93 right eyes and 108 left eyes. There were 93 right eyes and 108 left eyes. The diabetic retinopathy stages were as follows: stage Ⅳ in 24 eyes, stage Ⅴ in 78 eyes and stage Ⅵ in 99 eyes. The operation time was ranged from 1 to 4 hours, with an average of 2 hours. Among the 201 eyes, corneal epidermis were scraped on 25 eyes; 70 eyes were combined with cataract surgery; a laser photocoagulation count ＜1000 points was performed in 78 eyes, and ＞1000 points were performed in 123 eyes. Sixty-one eyes involved intravitreal silicone oil tamponade, 18 eyes involved intravitreal tamponade with C3F8, and 122 eyes were not involved with intraocular tamponade. Postoperative persistent intraocular hypertension was defined as an intraocular pressure (IOP) ≥21 mmHg (1 mmHg=0.133 kPa) after PPV with necessary treatment using IOP-lowering medications for ≥2 weeks. The diagnostic criteria for corneal epithelial defects were taken from the Expert Consensus on Clinical Diagnosis and Treatment of Corneal Epithelial Defect in China (2016). The corneal epithelial defect was diagnosed as PCED if it was treated with common methods such as a lacrimal substitute or corneal contact lens, but showed no improvement and no signs of healing for ≥2 weeks. The incidence of PCED after eye surgery was recorded and its related risk factors were analyzed. A multivariate logistic regression was used to analyze the risk factors for PCED, which were expressed as a odds ratio (OR) and a 95% confidence interval (CI). Results Of 201 eyes, 16 eyes (7.96%) presented with PCED after surgery and 185 eyes (92.04%) with no PCED. There was no significant difference in the age, sex, and eyes between the patients with or without PCED (χ2=6.548, 0.927, 0.044; P=0.011, 0.336, 0.833). A multivariate logistic regression showed that intraoperative epithelial debridement (OR=13.239, 95%CI 2.999−58.442, P=0.001), intraoperative treatment in combination with cataract surgery (OR=7.448, 95%CI 1.975−28.091, P=0.003), intravitreal tamponade with C3F8 (OR=11.344, 95%CI 2.169−59.324, P=0.004), and postoperative persistent intraocular hypertension (OR=10.462, 95%CI 2.464−44.414, P=0.001) were risk factors for PCED after PPV. Conclusion Intraoperative epithelial debridement, intraoperative treatment in combination with cataract surgery, intravitreal tamponade with C3F8, and postoperative persistent intraocular hypertension are risk factors for PCED in patients with PDR after PPV.