The amniotic membrane (AM) is used for engineering ex vivo tissue constructs used in ocular surface reconstruction. Epidermal growth factor (EGF) content of the AM is believed to play a key role in supporting corneal epithelial cell expansion on AM. This study investigated EGF content in AM in relation to intra- and inter-donor variations and the effect of processing and preservation (handling).Fifteen human AM, both fresh and handled, were analyzed for EGF gene and protein expression by real-time polymerase chain reaction and ELISA, respectively.EGF gene expression was predominantly seen in the AM epithelium (p<0.01). Similarly, EGF protein too was predominantly seen in the epithelial layer (p<0.01) for fresh and handled samples. EGF protein content varied between membranes (inter-donor) and at different sites within the same membrane (intra-donor). The highest EGF protein concentration was noted in the AM apical and mid-region epithelium. Significant EGF protein loss (p<0.01) was observed after handling.There is a considerable variation in EGF content between and within donors. This is further affected by handling of the AM. Such variations could affect the clinical efficacy of tissue constructs. Current use of AM for ex vivo expansion of epithelial cells is not standardized and remains an area of concern.
A bstract Background: The aim of the present study was to assess the relative effectiveness of tube surgery and cyclodiode laser in terms of achieving intraocular pressure control. Methods: A retrospective study was undertaken to compare patients undergoing double plate Molteno tube implantation with patients undergoing diode cyclophotocoagulation. Intraocular pressure (IOP) was documented at 7 days prior to surgery and postoperatively at various time points. Surgical success was defined as a final IOP between 6 (inclusive) and 21 mmHg (inclusive), without the use of topical medication, while ‘qualified’ success was defined as IOP within the same range with the use of topical medication. Results: Twenty‐eight diode patients and 26 tube patients were included for the study. An average follow up of 150 weeks (range = 21–322 weeks) was available. Mean preoperative IOP was 37 ± 12 mmHg for the tube group and 39 ± 16 mmHg for the diode group ( t = 0.51, P = 0.61). The final IOP was 17 ± 12 mmHg for the tube group and 21 ± 13 mmHg for the diode group ( t = 0.35, P = 0.73). Surgical success was achieved in 46% of tube eyes and 11% of diode eyes, while qualified success was achieved in 81% of tube eyes compared with 64% of eyes in the diode group. Two eyes which underwent diode became phthisical. Conclusions: IOP control may be achieved in a greater number of patients with tube surgery. The possible benefits of IOP control in diode patients need to be weighed against the risks of long‐term visual loss and the need for multiple re‐treatments in this group.
Purpose The extraembryonic coelom or spongy layer (SL) is the gelatinous, biochemically complex layer, which functions as a physical boundary between the vascular chorion and avascular amnion membrane (AM). SL often remains associated with the AM and is partially but variably removed during preparation for transplantation. We have shown that SL contains a similar profile of potentially beneficial factors as AM, but at considerably higher levels, which may explain in part the observed clinical variation. However, the effects of SL on ocular cell growth have yet to be established Methods Soluble proteins were extracted from freeze-dried SL to generate a soluble (sSL) and an insoluble structural SL fraction (iSL). Cultured corneal epithelial cells (CEC), keratocyte derived fibroblasts (KDF) and lymphocytes were stimulated with sSL and iSL fractions at varying dilutions, and the effects on cell proliferation (WST-1) and cytotoxicity were measured. Results At physiological concentration (16mg/ml) iSL killed all cell types within a few hours. Serial dilution of iSL (16mg/ml,8mg/ml,3.2mg/ml,0.32mg/ml) reduced rate of death, but death still occurred. Stimulation with sSL killed CEC at all concentrations, whilst the most dilute sSL fraction promoted KDF growth. Conclusion SL exerts a powerful cytotoxic effect on ocular and immune cells. However, the depot of factors contained within the SL may over time be released over time to promote cell growth. Therefore in the current situation where SL is typically ignored during AM preparation for transplantation, may have significant implications for the clinical efficacy of AM.
Purpose To report the outcome of orbital floor triamcinolone acetonide (OFTA) in refractory pseudophakic cystoid macular oedema(PCMO) and to determine the visual outcome in these patients Methods Six eyes of 6 patients with PCMO inadequately responsive to treatment combinations of topical steroidal and non-steroidal agents were retrospectively studied. All received 40mg (1ml) OFTA injection. Post-operative Visual acuity (VA), intraocular pressure (IOP) and OCT findings were assessed. Other potential complications were looked for retrospectively. Results The average age was 72 years(+/-12 years). OFTA was given, on average, 4 weeks after a diagnosis of PCMO was made (range 0-6 weeks) and treated with topical anti-inflammatory agent combinations. The mean follow-up was 11.0 months (range, 5-18), and the mean improvement of VA after OFTA was Snellen, (6/18-6/12). This was noticed at a mean of 12 weeks (range 4-72). At last follow-up, five eyes showed an improvement of two lines or more, while in one eye vision was maintained at 6/24 which developed diabetic maculopathy and required grid laser. None of the patients developed post-treatment raised IOP or lost vision. There was a significant reduction of retinal thickness and cystoid space height (P = 0.003). The dosage of topical steroids was reduced or stopped altogether in all 6 eyes. There were no cases of injection-related retrobulbar haemorrhage, cellulitis, or globe perforation Conclusion In cases of psuedophakic CMO, initial response to OFTA treatment was encouraging. Further larger long term studies are required to ascertain whether re-treatment is effective with subsequent orbital floor steroid injections. This is with a view to maintain the initial improvement.
Purpose The appearance of “de novo” tumors in adults receiving immunosuppressive treatment with tacrolimus in ocular inflammatory disorders has not been elucidated. Methods All 180 patients who received Tacrolimus as a steroid sparing agent for the management of high risk PKP, uveitis, scleritis or corneal stem cell allograft were studied. Tacrolimus treatment schedule, monitoring and duration of treatment was noted. Results During the last 8 years a total of 11 patients who had received Tacrolimus for their immunosuppresion developed a malignancy. Three patients developed cancer of prostate, bladder (1), bronchogenic carcinoma – squammous cell carcinoma (SCC) (1), cancer skin - (SCC) (1), breast cancer(2), recurrence of NHL (1) and recurrence of breast cancer (1) and large intestinal tumour (1). The primary diagnosis for the commencement of Tacrolimus included high-risk PKP (8), Scleritis (1), Panuveitis (1) and Wegeners Granulomatosis (1). The rate of increase of bladder cancer was 149.26 followed by NHL 82.33 and recurrent breast carcinoma 30.7. The rest of tumours had a rate of increase in the range of 10 - 20 fold. The average dose of Tacrolimus was 1954.37 mg SD +/- 2197 mg. Female/male ratio was 2:1. The mean age of patients was 73 years SD +/- 10.4 years. The mean follow-up time of tumour patients on treatment was 45 months with SD +/- 26 months and the mean duration of follow-up after diagnosis was 23 SD +/- 19 months. None of the eleven patients had any additional predisposing factors or were on any other carcinogenic agents which could affect their mortality by the cancer identified. Conclusion Long term immunosuppressive management of patients requires regular oncologic screening.
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