Purpose To investigate the benefit of intralesional injections of 5FU for the treatment of recurrent pterygium Methods Retrospective case note review of patients following surgical excision for pterygium identified 6 patients with recurrence treated with 0.1-0.2ml (2.5-5mg) of intralesional 5 FU post operatively. The time to recurrence, post recurrence injections, grade of pterygium pre and post treatment and time to regression were identified. Results The study group consisted of 5 male and 1 female patients, affecting 6 right eyes. 3 patients developed primary recurrence. The other 3 patients had previous surgery with recurrence followed by further surgical excision and repeat recurrence. The average grade of recurrence was 3.5. All 6 patients showed regression of the fibrovascular tissue following intralesional 5FU injections with an average dose of 0.1-0.2 ml (2.5-5mg). 5 patients required 3 injections or less for regression, whilst one patient required 14 post recurrence 5FU injections. The average grade following regression was 1.7. This level of regression was maintained at an average of 9 months follow-up. No complications of 5FU were reported. Conclusion The use of 5FU for the treatment of recurrent pterygium is safe and effective in limiting both progression and inducing regression of recurrent pterygium.
Purpose The eye despite being a delicate structure,has evolved many protective mechanisms.Antimicrobial peptides(AMPs) and Toll-like receptors(TLRs)have an important role in the ocular defense system.However during an infection or trauma, the innate defense system is breached.In this instance,keratocytes and fibroblasts aids in wound healing as well as provide a second line of defense.Little is known about AMP expression by TLR signalling pathways in corneal fibroblasts.This study therefore demonstrates the mRNA expression of both novel and known AMPs in various fibroblast phenotypes(myofibroblasts and keratocytes).In addition, AMPs were also shown in microbial ligand and/or pro-inflammatory cytokine stimulated human corneal fibroblast cultures (HCFC). Methods Fresh Keratocytes and HCFC were cultured and collected at different stages of growth and/or after stimulation with microbial ligands. Total RNA was extracted and cDNA was synthesized for semi-quantitative gene expression studies. Polymerase chain reaction (PCR) amplified product were analyzed on ethidium bromide stained 1% agarose gels. Results Constitutive expression of Thy-1(CD-90), vimentin, keratocan and alpha SMA was shown in all cultures.Keratocyte phenotype marker CD-34 showed decreased expression from 1st to 4th passage.Positive expression of Human beta defensin 109, Liver-expressed AMP(LEAP-2) and cathelicidin/LL-37 was demonstrated in cytokine and microbial ligand stimulated HCFC. Conclusion Reminiscent with previous studies,we have demonstrated the mRNA expression of novel and known AMPs in corneal fibroblast mediated by TLR dependent signalling mechanisms. This result warrants the further study to fully understand the role of AMPs in ocular wound healing.
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 The Limbal Epithelial Crypt (LEC) has been suggested to be the limbal epithelial stem cell niche and the limbus has been shown to demonstrate regional variations in association with the LEC. Our aims were to 1) to determine the existence of LEC in the infant age group and 2) compare and contrast the corneo-scleral tissue between infant and the adult by use of immunofluorescent imaging. Methods Donor eyes were processed and snap frozen in liquid nitrogen. Using a Leica cryostat machine, corneo-scleral sections were prepared and examined under microscopy for presence of LEC structures. Limbal sections were fixed, and stained with a range of current postulated markers of stem cells and cell differentiation by standard immunofluorescent imaging techniques. Results Immunostaining in the infant group demonstrates that both limbal and corneal epithelium consist of cells that are characteristic of cells with less differentiated nature. No classical adult LEC was discernable in corneo-scleral rims of a 2 year old or a 4-month old infant. However, adult LEC immunofluorescence, demonstrate similar characteristics to the infant limbus. Conclusion We hypothesize that the absence of classical adult LEC in infants suggests that LEC may arise during the natural human growth, and may act as a repository for limbal epithelial stem cells in the human adult.
Abstract Purpose To demonstrate in vitro the potential use of FDA‐approved PLGA poly(lactic‐co‐glycolic acid) microparticles to aid visualisation of vitreous during anterior vitrectomy following posterior capsule rupture and to compare with the use of triamcinolone suspension. Methods 10‐60um sized PLGA microparticles were fabricated using the single and/or double emulsion technique(s) and used i) untreated (ii) pre‐treatment with sodium hydroxide (iii) surface adsorption of a protein/synthetic peptide. Particle size, shape, morphology and surface topography were also assessed using scanning electron microscopy (SEM). The efficacy of these microparticles to enhance visualisation of vitreous against triamcinolone suspension was assessed using a simple in vitro set‐up exploiting human cadaveric vitreous. Results Unmodified PLGA displayed a degree of hydrophobicity and also a high rate of dispersion following repeated wash cycles. Interestingly, surface modified PLGA microparticles demonstrated a significant improvement in its 'vitreophilic' properties and were comparable to a triamcinolone suspension. Conclusion The use of modified biodegradable PLGA microparticles may represent a novel method of visualising vitreous and aiding anterior vitrectomy. This method may provide a distinct alternative for the visualisation of vitreous without the pharmacological effects of triamcinolone. Furthermore, such a technique allows the incorporation of drugs, growth factors and/or antibiotics within the microparticles which may be desirable in complicated cataract extraction. Animal models are required to assess in vivo biocompatibility of this method.
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