Factors correlated with visual outcomes at two and four years after vitreous surgery for proliferative diabetic retinopathy.

Proliferative diabetic retinopathy (PDR) is the most severe case of diabetic retinopathy that can cause visual impairment. This study aimed to reveal the factors correlated with better postoperative visual acuity after a long follow-up in patients who underwent vitrectomy for PDR. We retrospectively analyzed the data set including systemic findings, ocular findings, and surgical factors from registered patients who could be completely followed up for 2 or 4 years after vitrectomy. We ultimately enrolled 128 eyes from 100 patients who underwent vitrectomy for PDR between January 2008 and September 2012 and were followed up for >2 years. Among them, 91 eyes from 70 patients could be followed up for 4 years. Factors related to the postoperative visual acuity of ≥20/40 and ≥20/30 after 2 and 4 years were investigated by logistic regression analysis. Better postoperative visual acuity correlated with the following factors: no rubeosis iridis ([≥20/40 at 2 years; odds ratio {OR}, 0.068; 95% confidence interval {CI}, 0.012-0.39; P = 0.003], [≥20/30 at 2 years; OR, 0.07; 95% CI, 0.01-0.40; P = 0.03], [≥20/30 at 4 years; OR, 0.078; 95% CI, 0.006-0.96; P = 0.04]), no fibrovascular membrane [(≥20/40 at 2 years; OR, 0.22; 95% CI, 0.061-0.81; P = 0.02), (≥20/40 at 4 years; OR, 0.26; 95% CI, 0.07-0.94; P = 0.04), (≥20/30 at 4 years; OR, 0.14; 95% CI, 0.04-0.52; P = 0.004)], existing vitreous hemorrhage (≥20/30 at 2 years; OR, 9.55; 95% CI, 1.03-95.27; P = 0.04), and no reoperation ([≥20/40 at 4 years; OR, 0.15; 95% CI, 0.03-0.78; P = 0.02], [≥20/30 at 4 years; OR, 0.06; 95% CI, 0.07-0.54; P = 0.01]). Treatment provision before disease severity and treatment without complications were associated with good postoperative visual acuity.

Cotylenin A inhibits cell proliferation and induces apoptosis and PAX6 mRNA transcripts in retinoblastoma cell lines.

PURPOSE: Retinoblastoma, a childhood cancer of the retina, is caused by inactivation of the tumor suppressor gene retinoblastoma (RB). Cotylenin A (CN-A), a novel fusicoccane-diterpene glycoside, accelerates the differentiation of several types of myeloid cell lines and is a candidate for a new type of anticancer therapeutic agent with this effect. However, whether CN-A has the same effect on retinoblastoma cells is unknown. We studied the response of two retinoblastoma cell lines, Y-79 and WERI-Rb-1, to CN-A. METHODS: We studied the response of two retinoblastoma cell lines to CN-A with respect to cell growth, apoptosis, morphology, mRNA, protein expression analysis of specific genes (N-myc, cyclin-dependent kinase inhibitor 1A [P21], paired box gene 6 [PAX6], and rhodopsin [RHO]), and activity of three PAX6 promoters (P0, P1, and Palpha). RESULTS: CN-A inhibited cell proliferation and induced apoptosis via caspase activity in the two retinoblastoma cell lines. In addition, CN-A induced mRNA expression of P21, PAX6, and RHO and protein expression of P21. In Y-79 cells, PAX6 P1 promoter was activated by CN-A. In WERI-Rb-1 cells, PAX6 P0, P1, and Palpha promoter were activated by CN-A. CN-A decreased mRNA and protein expression of N-myc in two retinoblastoma cell lines. CONCLUSIONS: The responses of retinoblastoma cells to CN-A include inhibition of cell growth, induction of apoptosis, and the potential to change neuroblastoma characteristics of retinoblastoma cells.

Steroid eye drop treatment (difluprednate ophthalmic emulsion) is effective in reducing refractory diabetic macular edema.

PURPOSE: To evaluate the efficacy of treatment of refractory diabetic macular edema (DME) after vitrectomy with difluprednate ophthalmic emulsion 0.05% (Durezol(TM)), and to compare this treatment with sub-Tenon's injection of triamcinolone (STTA). METHODS: This study enrolled patients with refractory diabetic macular edema that persisted despite pars plana vitrectomy in our clinic. In all subjects, more than 3 months had passed since prior treatment. Eleven eyes in ten subjects were treated with STTA (STTA group), and 11 eyes in seven subjects were treated with difluprednate ophthalmic emulsion 0.05% (Durezol(TM), Sirion Therapeutics Inc., USA) 4 times daily for the first month and then twice daily for 2 months (eye drop group). RESULTS: In the eye drop group, mean VA (+/- SD) was 0.67 +/- 0.35 logMAR and mean retinal thickness was 500.6 +/- 207.7 mum at baseline. After 3 months of treatment, mean VA was 0.67 +/- 0.29 and mean retinal thickness had decreased to 341.2 +/- 194.8 mum. The mean minimum value of RT during the treatment period was 300.6 +/- 123.2 mum, and significantly lower than that at baseline (Mann-Whitney U test: P = 0.003). In the STTA group, mean VA (+/- SD) was 0.67 +/- 0.35 logMAR, and mean retinal thickness was 543.3 +/- 132.6 mum at baseline. After 3 months of treatment, mean VA was 0.49 +/- 0.67, and mean retinal thickness had decreased to 378.6 +/- 135 mum. The mean minimum value of RT during the treatment period was 349.9 +/- 113.8 mum, and significantly lower than at baseline (Mann-Whitney U test: P = 0.003). The rate of effective improvement in RT did not differ between the eye drop group (73%) and STTA group (84%) (Fisher's exact test: P = 1). CONCLUSIONS: Comparable improvements of retinal thickness were observed in the STTA and eye drop groups. Instillation of difluprednate ophthalmic emulsion 0.05% is a safe and effective treatment that does not require surgical intervention and does not produce severe side-effects.

Two-year visual outcomes after photodynamic therapy in age-related macular degeneration patients with or without polypoidal choroidal vasculopathy lesions.

PURPOSE: To describe the visual outcomes 2 years after photodynamic therapy in Japanese patients with age-related macular degeneration (AMD) with or without polypoidal choroidal vasculopathy (PCV) lesions. METHODS: Sixty-three eyes of 63 consecutive patients with AMD or AMD + PCV who underwent photodynamic therapy were included in this study. Fluorescein and indocyanine green angiography were performed to diagnose AMD and AMD + PCV. Change in mean visual acuity and recurrence of active lesion during the follow-up period up to 2 years were assessed. RESULTS: Patients with typical AMD maintained visual acuity for 2 years after photodynamic therapy. For patients with AMD + PCV, the visual acuity was maintained during the first year but started decreasing by 0.09 logarithm of the minimum angle of resolution units per 3 months (95% confidence intervals [CI], 0.06-0.14) after 1 year. Moreover, patients with AMD + PCV had 82% higher risk of a recurrence of active lesions for each increase in 3 months of follow-up time after 1 year; this suggested that the risk of recurrence had increased later in follow-up after 1 year. Recurrence of active PCV lesions and massive subretinal hemorrhages were the main reasons for the late worsening of visual acuity. CONCLUSION: The visual acuity after photodynamic therapy in AMD patients was maintained for 2 years after the initial treatment. Patients with AMD + PCV had stable visual outcome within 1 year but not after 1 year; there are risks of late recurrences and massive hemorrhages after 1 year in patients with AMD + PCV.

Long-term follow-up results of pars plana vitrectomy for diabetic macular edema.

PURPOSE: To analyze the long-term effects of pars plana vitrectomy for diabetic macular edema. METHODS: Sixty-nine patients (73 eyes) with diabetic macular edema were examined retrospectively after pars plana vitrectomy. The mean follow-up time was 24.6 +/- 7.3 months. RESULTS: The postoperative best-corrected visual acuity (BCVA) at 12 months was significantly better than the preoperative BCVA, and the improvement was maintained for at least 24 months. The mean central retinal thickness (RT) at 3 months after surgery was significantly thinner than the preoperative central RT, and was maintained for at least 24 months in the cases followed for this period. The postoperative BCVA was significantly worse than the preoperative BCVA at 2 years after surgery in eyes with a preoperative BCVA of < or =0.05. In addition, there was lower probability that the postoperative BCVA would be > or =0.5 in eyes with a preoperative BCVA of <0.3. CONCLUSIONS: The results indicate that the improvement in the BCVA is attained by 12 months postvitrectomy, and is maintained for at least 24 months. The reduction in RT can be maintained for up to 24 months. The results also indicate that vitrectomy should be performed when the preoperative BCVA is >0.05 at worst.

[Long-term results of grid pattern photocoagulation for diffuse diabetic macular edema].

PURPOSE: To investigate the long-term results of grid pattern photocoagulation for diffuse diabetic macular edema. SUBJECTS AND METHODS: In 45 cases, 56 eyes foveal thickness and visual outcome were observed 12 months or more after grid pattern photocoagulation. RESULTS: Compared with preoperative values, average foveal thickness decreased significantly during the initial 3 postoperative months and this reduction was maintained through 30 postoperative months. Foveal thickness was reduced by more than 20% in 66% of eyes. The final logarithm of the minimum angle of resolution (logMAR) visual acuity improved 0.2 or more in 41% of eyes. There was a significant correlation between preoperative and postoperative visual acuity. A final visual acuity of 0.5 or better was obtained in 80% of eyes with a preoperative visual acuity of 0.4 or better. There were no eyes with visual loss due to atrophic creep in this study. CONCLUSION: Grid pattern photocoagulation for diffuse diabetic macular edema showed long-term effectiveness in reducing foveal thickness postoperatively. It will be possible to achieve a final visual acuity of 0.5 or better in a high percentage of cases, if grid pattern photocoagulation is performed for those with relatively good visual acuity. The foveal thickness measurement was useful for quantitative evaluation of photocoagulation for diabetic macular edema.

Early postoperative retinal thickness changes and complications after vitrectomy for diabetic macular edema.

PURPOSE: To determine the early postoperative changes in retinal thickness and complications after pars plana vitrectomy for diabetic macular edema. DESIGN: Consecutive interventional case series. METHODS: Studied retrospectively, pars plana vitrectomy was performed on 65 consecutive eyes of 63 patients with diabetic macular edema. The follow-up interval ranged from 6 to 36 months (12.6 +/- 7.4 months [mean +/- standard deviation (SD)]). The indications of pars plana vitrectomy in this study were (1) diffuse diabetic macular edema, (2) preoperative visual acuity less than 20/40, and (3) noneffective macular photocoagulation therapy. Preoperative and postoperative examinations by stereoscopic biomicroscopy, color fundus photography of the macula and optical coherence tomography (OCT) were performed on all eyes. Preoperatively, direct photocoagulation to microaneurysms in the macula had been performed in 48 eyes, and focal/grid photocoagulation had been performed in five eyes. Preoperative examination showed that epiretinal membranes were observed in 20 eyes, cystoid macular edema in 40 eyes, and 23 eyes had a complete posterior vitreous detachment (PVD). Epimacular membranes, removed during surgery, were examined histopathologically. RESULTS: The postoperative mean best-corrected visual acuity (logarithm of the minimum angle of resolution [logMAR] = 0.696 +/- 0.491 [mean +/- SD]) was significantly better than the preoperative mean best-corrected visual acuity (0.827 +/- 0.361; P <.0001; Wilcoxon signed-rank test). The final visual acuity improved by 2 or more lines in 32 of 65 eyes (45%), remained unchanged in 32 of 65 eyes (49%), and exacerbated after the surgery in 4 of 65 eyes (6%) due to neovascular glaucoma (2 eyes) and residual cystoid macular edema (2 eyes). The postoperative foveal retinal thickness (224.9 +/- 116.9 microm) at the last visit was significantly thinner than the preoperative foveal retinal thickness (463.7 +/- 177.3 microm; P <.0001; Wilcoxon signed-rank test). The foveal retinal thickness did not decrease linearly but fluctuated: The mean postoperative retinal thickness had decreased significantly 7 days after surgery, then remained unchanged for approximately 1 month, and thereafter gradually decreased until 4 months. The intraoperative and postoperative complications included peripheral retinal tear in 3 of 65 (4.6%) eyes, postoperative rhegmatogenous retinal detachment in 1 of 65 (1.5%) eyes, neovascular glaucoma in 3 of 65 (5%) eyes, recurrent vitreous hemorrhage in 1 of 65 (1.5%) eyes, hard exudates in the center of the macula in 3 of 56 (4.6%) eyes, postoperative epiretinal membrane formation in 9 of 65 (13.8%) eyes, and a lamellar macular hole in 1 of 65 (1.5%) eyes. CONCLUSIONS: Vitrectomy for diabetic macular edema is an effective procedure for reducing the edema and improving visual acuity. Because the postoperative reduction in retinal thickness is not complete until 4 months, the assessment of vitrectomy on foveal thickness should not be made until this time. In addition, there are severe complications from vitrectomy for diabetic macular edema, and careful preoperative and postoperative examinations and surgical methods are required.

Screening and follow-up of diabetic retinopathy using a new mosaic 9-field fundus photography system.

AIM: To evaluate the clinical usefulness of a newly developed fundus photographing system and assess its applicability to telemedicine. METHODS: Nine overlapping 45 degrees fundus photographs were taken by a new camera equipped with nine internal fixation targets to provide standardized 9-field photographs. The digitally stored images were either edited in 3x3 form or reconstructed as collage (9F) and compared to the ophthalmological examination (OP) and single-field non-mydriatic photography (SC). In telemedicine, 9-field images derived from 61 adolescent diabetics were sent to ophthalmologists over an analog phone line. RESULTS: The sensitivities of the examinations by 9F without and with mydriasis (78 and 82%) were equivalent to OP (84%) and superior to SC (64%). The diagnosis of severity by 9F was also comparable to those by OP, whereas SC tended to underestimate the severity. An average of 1 min 19 s was required to send one edited 9-field photography (average size 259+/-30 KB) over the Internet. Twelve of these eyes were diagnosed as diabetic retinopathy on a desktop monitor whereas SC gave only seven. CONCLUSION: This new 9-field fundus photography system can be appropriate for the screening and follow-up of diabetic retinopathy in adult and adolescent diabetic subjects, especially when applied to telemedicine over the Internet.

Expression of hyaluronan synthase in intraocular proliferative diseases: regulation of expression in human vascular endothelial cells by transforming growth factor-beta.

PURPOSE: To investigate the role of hyaluronan (HA) and elucidate the mechanisms that regulate the expression of hyaluronan-synthesizing enzymes in vascular endothelial cells (VECs) in intraocular proliferative diseases. METHODS: Cultured VECs were used. Hyaluronan synthase (HAS) expression was determined on the mRNA products obtained by reverse transcription polymerase chain reaction (RT-PCR). The effect of transforming growth factor-beta(1)(TGF-beta(1)) and/or platelet-derived growth factor-BB (PDGF-BB) on HAS expression was examined by quantitative RT-PCR and Western blot analysis. HAS expression in intraocular proliferative membranes was observed by immunohistochemistry. RESULTS: Cultured VECs expressed the three HAS isoforms. Stimulation of VECs with TGF-beta(1) induced a marked increase in the expression level of HAS2 mRNA and protein. The stimulatory effect of PDGF-BB was less potent. A synergistic or additive effect between TGF-beta(1) and PDGF-BB-induced HA synthesis was not observed. Furthermore, HAS1 and HAS2 exhibited differential expression in VECs and non-VECs populating intraocular proliferative membranes. CONCLUSIONS: The expression of each HAS isoform is regulated differently by growth factors and cytokines in VECs. Importantly, HA-synthesizing enzymes were expressed in cells populating proliferative membranes obtained from eyes of patients with proliferative vitreoretinal diseases, and thus may be key molecules in the events that control progression of the proliferative diseases.

Monocyte chemotactic protein-1 levels in the vitreous of patients with proliferative vitreoretinopathy.

PURPOSE: To assess the potential role of monocyte chemotactic protein-1 (MCP-1) in the pathogenesis of proliferative vitreoretinopathy (PVR) and to investigate its possible interaction with the macrophage migration inhibitory factor (MIF). METHODS: We assayed MCP-1 and MIF levels in the vitreous samples of 85 consecutive patients with PVR (29 eyes), rhegmatogenous retinal detachment (RRD; 22 eyes), and macular hole or idiopathic epimacular membrane (controls; 34 eyes), by enzyme-linked immunosorbent assay. RESULTS: Vitreous levels of MCP-1 were 1760.7 +/- 471.3 pg/mL (mean +/- SD) in PVR patients, 1200.4 +/- 579.8 pg/mL in RRD patients, and 436.3 +/- 286.1 pg/mL in the controls. Vitreous MCP-1 levels in PVR patients were significantly higher than those in RRD patients and in the controls (P <.0001, respectively). MCP-1 levels in grade C of PVR (1883.7 +/- 479.5 pg/mL) were significantly greater than those in grade D (1437.8 +/- 258.8 pg/mL) (P =.0112). Vitreous concentrations of MCP-1 had no correlation with those of MIF. CONCLUSIONS: The results indicate the possibility that MCP-1 may have a role mainly in the early stage of PVR and that the role of MCP-1 in PVR may differ from that of MIF.

Long-term outcomes of 3 surgical adjuvants used for internal limiting membrane peeling in idiopathic macular hole surgery.

PURPOSE: Indocyanine green (ICG), an adjuvant used for peeling of the internal limiting membrane (ILM) during vitreous surgery for idiopathic macular hole (MH), has been reported to be toxic, possibly affecting postoperative visual acuity. We compared the long-term outcomes (within 2 years) of brilliant blue G (BBG), ICG, and triamcinolone acetonide (TA). PATIENTS AND METHODS: This study involved 97 eyes of 94 patients who underwent vitreous surgery for MH at the Yamagata University Hospital between June 2002 and November 2010. The surgical adjuvants used were BBG for 15 eyes, ICG for 61 eyes, and TA for 21 eyes. We compared the postoperative visual acuities, initial closure rates, final closure rates, and complications of the 3 groups. RESULTS: In all 3 groups, the visual acuity significantly improved after surgery. The magnitude of the improvement at 2 years after surgery was significantly better in the BBG group than in the ICG group (Mann-Whitney test, P = 0.020). The postoperative visual acuity did not significantly differ between the BBG and TA groups (P = 0.627) or between the ICG and TA groups (P =0 .137). Thus, the surgery using BBG resulted in a significantly better outcome in visual acuity than did the surgery using ICG. The 3 groups did not differ in initial or final closure rates or in incidence of complications. CONCLUSION: Analysis of the long-term outcomes of vitreous surgeries provided evidence that BBG is a useful adjuvant for ILM peeling.

Three-dimensional imaging of the inner limiting membrane folding on the vitreomacular interface in diabetic macular edema.

PURPOSE: To report the findings of fine folds on the retina obtained by spectral-domain optical coherence tomography (OCT). METHODS: A retrospective non-comparative case series; 26 eyes of diabetic macular edema (DME) patients who underwent vitrectomy were observed using three-dimensional (3D) images of OCT preoperatively and postoperatively. The specimens were investigated immunohistochemically. RESULTS: Using only tomography, non-tractional vitreoretinal interfaces were observed in 15 eyes and tractional vitreoretinal interfaces in the other 11 eyes. Using 3D imaging, we observed fine folds in 11 eyes among 15 cases showing non-tractional interfaces. Based on these findings, the state of the vitreoretinal interface was classified into 3 patterns. Group 1, both tomography and 3D imaging showed smooth retinal surfaces. Group 2, tomography showed a smooth retinal surface, but 3D imaging showed fine folds on the retina. Group 3, both tomography and 3D imaging showed a tractional vitreoretinal interface with an obvious epiretinal membrane and/or taut posterior vitreous cortex. The fine folds in group 2 disappeared and macular edema improved after inner limiting membrane (ILM) peeling, and the CRT of groups 2 and 3 reduced significantly. The fine folds were confirmed to involve the ILM because type IV collagen expression was detected in the surgically obtained specimens. CONCLUSION: We observed tangential fine folds of the ILM. These were detected by using only 3D imaging, and might be useful for investigating the optimal indication of vitrectomy for DME.

Prevalence and risk factors for diabetic maculopathy, and its relationship to diabetic retinopathy in elderly Japanese patients with type 2 diabetes mellitus.

AIMS: To determine the prevalence of diabetic retinopathy and diabetic maculopathy in Japanese patients older than 65 years-of-age with type 2 diabetes mellitus. In addition, to determine the relationship between the severity of retinopathy and maculopathy, and the risk factors for these conditions in Japanese patients with the same characteristics. METHODS: This was a cross-sectional study carried out at the enrolment of patients who participated in a randomized controlled trial. A total of 960 eyes of 960 Japanese patients with type 2 diabetes who were ≥ 65 years-of-age were analyzed. RESULTS: Our data showed that there was a correlation between the severity of retinopathy and the severity of maculopathy. The risk factors for the severity of retinopathy were different from the risk factors for the severity of maculopathy. The age, duration of diabetes, systemic pulse pressure, fasting insulin, insulin treatment of diabetes, high-density lipoprotein cholesterol, microalbumin-to-creatinine ratio and history of cerebrovascular disease all contributed significantly to the severity of retinopathy. The duration of diabetes, insulin treatment and microalbumin-to-creatinine ratio were correlated with the severity of maculopathy. CONCLUSIONS: The risk factors related to diabetic retinopathy and maculopathy in Japanese patients with type 2 diabetes mellitus aged ≥ 65 years were different from that in other countries. Our data also showed that the certain risk factors for retinopathy differ from those associated with maculopathy.

Treatment of diffuse diabetic macular oedema using steroid eye drops.

PURPOSE: To evaluate the efficacy of treatment of diffuse diabetic macular oedema (DME) with difluprednate ophthalmic emulsion 0.05% (Durezol™) in eyes before vitrectomy. METHODS: This study enrolled patients with diffuse DME for whom more than 3 months had passed since prior treatment. Nineteen eyes in 15 subjects were treated with difluprednate ophthalmic emulsion 0.05% four times daily for the first month and then twice daily for 2 months (treatment group). As a control group, 22 eyes in 11 subjects with DME were selected from subjects who underwent the steroid responder test. RESULTS: In the treatment group, the mean visual acuity (VA) (±SD) was 0.38 ± 0.25 logMAR and mean retinal thickness was 461.1 ± 109.9 µm at baseline. After 1 month of treatment, the mean VA had improved to 0.29 ± 0.25 (Wilcoxon rank-sum test, p = 0.30), while mean retinal thickness had decreased to 372.1 ± 70.0 µm (p = 0.006). The rate of effective improvement in retinal thickness was 42% and that of VA was 26%. In the control group, changes in neither VA nor retinal thickness were significant. CONCLUSIONS: Eye drop therapy using difluprednate ophthalmic emulsion 0.05% is a useful and effective treatment modality without surgical intervention or severe side-effects.

Difluprednate ophthalmic emulsion 0.05% as adjunctive treatment to panretinal photocoagulation for proliferative diabetic retinopathy with clinically significant macular edema.

PURPOSE: The purpose of this study was to evaluate the efficacy of topical difluprednate ophthalmic emulsion in controlling progressive diabetic macular edema after panretinal photocoagulation. METHODS: This was a case report of two patients with proliferative diabetic retinopathy combined with diabetic macular edema who underwent panretinal photocoagulation combined with use of a topical difluprednate ophthalmic emulsion. RESULTS: In the Case 1, retinal thickness was decreased 29% 1 month after the start of difluprednate treatment and best-corrected visual acuity was improved 20/40 to 20/25. In Case 2, retinal thickness was decreased 43% after 1 month, and best-corrected visual acuity was improved 20/100 to 20/60 after 3 months. During the follow-up period, elevation of intraocular pressure, ocular infection, and progression of cataract were not detected, though superficial punctuate keratitis was observed in one case. CONCLUSION: Topical difluprednate ophthalmic emulsion was one of the possible choices to treat progressive diabetic macular edema after panretinal photocoagulation. It is mandatory to evaluate the effects and safety in further studies including many cases.

Cloning and characterization of human vitreous tissue-derived cells.

PURPOSE: Previously, we established a porcine vitreous tissue-derived hyalocyte cell line (PH5) and investigated the regulation of hyaluronan synthesis in these cells by cytokines. The objective of the current study was to establish human vitreous tissue-derived cells and to compare their characteristics with those of PH5 cells. METHODS: Human vitreous specimens from two patients were cultured in the presence of 10% foetal bovine serum and immortalized by infection with human papilloma virus 16 genes E6 and E7. We used reverse transcription polymerase chain reaction (RT-PCR) to analyse and compare the expression profiles for several genes in the human vitreous tissue-derived cells and PH5 cells. To investigate the regulation of hyaluronan production in response to cytokine stimulation, the expression of hyaluronan synthase isoforms was examined using RT-PCR, and hyaluronan production was measured using enzyme-linked immunosorbent assay (ELISA). RESULTS: Two types of cells, HV64 and HV65, were derived from human vitreous tissue. The HV64 and HV65 cell-doubling times were 58 r and 76 hr, respectively. The cells expressed messenger RNA (mRNAs) encoding collagen type I α1 (COL1A1), collagen type II α1 (COL2A1), CD11b, CD14, CD68, CD204 and CD206 but did not express mRNA for glial fibrillary acidic protein (GFAP). Cytokine stimulation did not induce the expression of hyaluronan synthase mRNA or the production of hyaluronan. In contrast, mRNAs for GFAP and hyaluronan synthase-2 were expressed in the porcine PH5 cells, and treatment with transforming growth factor-ß1 and/or platelet-derived growth factor-BB induced the production of hyaluronan in PH5 cells. CONCLUSION: The new human vitreous tissue-derived cells have macrophage-like characteristics and are different from our previously developed porcine hyalocyte cells. These human vitreous tissue-derived cells might be useful for studies of human intraocular diseases.

Hyaluronan production and hyaluronan synthase expression in three human conjunctiva-derived cell strains.

PURPOSE: The conjunctiva maintains the health of the ocular surface by protecting the eye from pathogen invasion, injury, and dryness. In this study, we investigated the regulation of hyaluronan (HA) synthesis by cytokines in conjunctiva-derived cells. METHODS: Cultured primary cells derived from human conjunctivas that had been removed as surgical specimens were transfected with an immortalizing gene (human papilloma virus 16 E6/E7). To compare the biological features of the primary and immortalized cells, we assessed their morphological features and gene expression patterns. We also examined the effects of inflammatory cytokines on hyaluronan synthase (HAS) expression and HA production. RESULTS: Three conjunctiva-derived cell strains were established and could be passaged up to 15 times. All strains expressed ß2MG and KRT13 transcripts, highly expressed in conjunctival epithelial cells. HA production and expression of the three HAS isoforms were detected in the cell strains; however, cytokine treatment had no significant effect on HA production or HAS isoform expression. CONCLUSIONS: We succeeded in deriving three human cell strains from conjunctival tissue. In the conjunctiva-derived cell strains, HA production and HAS mRNA expression were stable and were not changed by either TGF-ß or PDGF-BB.