The Development of an Antimicrobial Contact Lens - From the Laboratory to the Clinic.

Contact lens wear is generally safe and provides excellent vision. However, contact lens wear is often associated with the risk of developing ocular surface infection and inflammation, and in severe cases, the infection can result in loss of vision. Antimicrobial peptide-coated contact lenses have been made to help reduce the incidence of infection and inflammation. This paper reviews the research progress from conception, through the laboratory and preclinical tests to the latest information on clinical testing of an antimicrobial contact lens. We provide insights into the pathways followed and pitfalls that have been encountered. The journey has not always been linear or smooth, but has resulted in some of the first published clinical testing of antimicrobial peptide-coated contact lenses in humans. We hope this may help lead to the development and commercialisation of antimicrobial contact lenses in the future.

Protective Action of Linear Polyethylenimine against Staphylococcus aureus Colonization and Exaggerated Inflammation in Vitro and in Vivo.

Increased evolution of multidrug resistant pathogens necessitates the development of multifunctional antimicrobials. There is a perceived need for developing new antimicrobials that can interfere with acute inflammation after bacterial infections. Here, we investigated the therapeutic potential of linear polyethylenimine (LPEI) in vitro and in vivo. The minimum inhibitory concentration of LPEI ranged from 8 to 32 µg/mL and elicited rapid bactericidal activity against clinical isolates of meticillin-resistant Staphylococcus aureus (MRSA). The polymer was biocompatible for human cultured ocular and dermal cells. Prophylactic addition of LPEI inhibited the bacterial colonization of human primary dermal fibroblasts (hDFs). In a scratch wound cell migration assay, LPEI attenuated the migration inhibitory effects of bacterial secretions. The polymer neutralized the cytokine release by hDFs exposed to bacterial secretions, possibly by blocking their accessibility to host cell receptors. Topical instillation of LPEI (1 mg/mL) was noncytotoxic and did not affect the re-epithelialization of injured porcine cornea. In a prophylactic in vivo model of S. aureus keratitis, LPEI was superior to gatifloxacin in terms of reducing stimulation of cytokines, corneal edema, and overall severity of the infection. These observations demonstrate therapeutic potential of LPEI for antimicrobial prophylaxis.

Prophylactic Vancomycin Drops Reduce the Severity of Early Bacterial Keratitis in Keratoprosthesis.

BACKGROUND: Artificial cornea transplantation, keratoprosthesis, improves vision for patients at high risk of failure with human cadaveric cornea. However, post-operative infection can cause visual loss and implant extrusion in 3.2-17% of eyes. Long-term vancomycin drops are recommended following keratoprosthesis to prevent bacterial keratitis. Evidence, though, in support of this practice is poor. We investigated whether prophylactic vancomycin drops prevented bacterial keratitis in an animal keratoprosthesis model. METHODOLOGY: Twenty-three rabbits were assigned either to a prophylactic group (n = 13) that received vancomycin 1.4% drops 5 times/day from keratoprosthesis implantation to sacrifice, or a non-prophylactic group (n = 10) that received no drops. All rabbits had Staphylococcus aureus inoculation into the cornea at 7-12 days post-implantation and were sacrificed at predetermined time-points. Prophylactic and non-prophylactic groups were compared with slit-lamp photography (SLP), anterior segment optical coherence tomography (AS-OCT), and histology, immunohistochemistry and bacterial quantification of excised corneas. Corneal vancomycin pharmacokinetics were studied in 8 additional rabbits. RESULTS: On day 1 post-inoculation, the median SLP score and mean±SEM AS-OCT corneal thickness (CT) were greater in the non-prophylactic than the prophylactic group (11 vs. 1, p = 0.049 and 486.9±61.2 vs. 327.4±37.1 µm, p = 0.029 respectively). On days 2 and 4, SLP scores and CT were not significantly different. Immunohistochemistry showed a greater CD11b+ve/non-CD11b+ve cell ratio in the non-prophylactic group (1.45 vs. 0.71) on day 2. Bacterial counts were not significantly different between the two groups. Corneal vancomycin concentration (2.835±0.383 µg/ml) exceeded minimum inhibitory concentration (MIC) for Staphylococcus aureus only after 16 days of vancomycin drops. Two of 3 rabbits still developed infection despite bacterial inoculation after 16 days of prophylactic drops. CONCLUSIONS: Prophylactic vancomycin drops provided short-term benefit, but did not prevent infection. Achieving MIC in the cornea was not sufficient to prevent Staphylococcus aureus keratitis. Patients should continue to be counselled regarding the risk of infection following keratoprosthesis.

[Management of peripheral facial nerve palsy in children]. / Prise en charge de la paralysie faciale périphérique chez l'enfant.

Peripheral facial nerve palsy may (secondary) or may not have a detectable cause (idiopathic facial palsy or Bell's palsy). Idiopathic facial palsy is the common form of facial palsy. It remains diagnosis by exclusion. The prognosis is more favourable in children than in adults. We present current diagnostic procedures and recommendations regarding treatment in children.

Identification and susceptibility to multipurpose disinfectant solutions of bacteria isolated from contact lens storage cases of patients with corneal infiltrative events.

Corneal infiltrative events (CIEs) are being reported with increasing frequency in lens wearers and may be related to specific multipurpose disinfecting solution (MPDS), contact lens type or bacterial bio-burden. Here, the efficacy of MPDS's against bacteria from contact lens storage cases (CLSC) of patients with CIEs was investigated. Eighteen CLSC from patients with CIEs were cultured. All reported using the same MPDS based on PQ-1+Aldox+nonanoyl-EDTA prior to experiencing CIEs. Bacteria were identified and tested for sensitivity to MPDS-1 and three other MPSDs. 16/18 CLSC (89%) contained bacterial counts of ≥10(4)-10(8)/mL. Achromobacter spp. was most frequently identified and was found in 11/18 cases (61%). This was followed by 4/18 (22%) Stenotrophomonas maltophilia, 3/18 (17%) Serratia marcescens, 3/18 (17%) Delftia spp., 2/18 (11%) Elizabethkingia spp., 2/18 (11%) Chryseobacterium indologenes and 1/18 Sphingobacterium spiritivorum. Acanthamoeba was not isolated. All of the Achromobacter strains were resistant to MPDS-1 with <1log10 kill up to 14 days exposure and the solution also showed reduced efficacy against the other isolates at the manufacturer's recommended disinfection time of 6h. Two strains of S. maltophilia and Delftia spp. grew in the solution over 14 days. Factors responsible for causing adverse events such as CIEs in contact lens wearers remain unclear. However, the presence of significant bio-burden in the contact lens storage case and lens may initiate an immunological response resulting in CIEs either directly or through the release of endotoxins (e.g. lipopolysaccharides) from the bacterial outer cell membrane.

Inhibition of corneal inflammation following keratoplasty by birch leaf extract.

The objective of this study was to determine the effect of birch leaf (Betula pendula) extract (BPE) on corneal inflammation following keratoplasty in the rat model. T cells were stimulated in vitro in the presence of BPE. Proliferation, activation phenotype and the number of apoptotic/necrotic cells in cell culture were analyzed by flow cytometry. Corneal transplantation was performed between Fisher and Lewis rats. Recipient rats were either treated with cyclosporine A at a low dosage (Low-dose CsA=LDCsA) or received LDCsA in combination with BPE (2×1ml/day). Clinical signs for corneal inflammation and rejection time points were determined. Infiltrating leukocytes were analyzed histologically. BPE specifically inhibited T cell proliferation in vitro by inducing apoptosis. The phenotype was not affected. In vivo, BPE significantly delayed the onset of corneal opacification (p<0.05). The amount of infiltrating CD45(+) leukocytes and CD4(+) T cells (p<0.001) was significantly reduced by BPE, whereas infiltration of CD163(+) macrophages was not significantly different between the two groups. BPE selectively induces apoptosis of activated T cells. Accordingly, BPE treatment significantly reduces infiltrating T cells and subsequent corneal opacification following keratoplasty. Our findings suggest BPE as a promising anti-inflammatory drug to treat corneal inflammation.

Exacerbated corneal inflammation and neovascularization in the HO-2 null mice is ameliorated by biliverdin.

Heme oxygenase (HO-1 and HO-2) represents an intrinsic cytoprotective and anti-inflammatory system based on its ability to modulate leukocyte migration and to inhibit expression of inflammatory cytokines and proteins. HO-2 deletion leads to unresolved corneal inflammation and chronic inflammatory complications including ulceration, perforation and neovascularization. We examined the consequences of HO-2 deletion on hemangiogenesis and lymphangiogenesis in the model of suture-induced inflammatory neovascularization. An 8.0 silk suture was placed at the corneal apex of wild type and HO-2 null mice. Neovascularization was assessed by vital microscopy and quantified by image analysis. Hemangiogenesis and lymphangiogenesis were determined by immunofluorescence staining using anti-CD31 and anti-LYVE-1 antibodies, respectively. Inflammation was quantified by histology and myeloperoxidase activity. The levels of HO-1 expression and inflammatory cytokines were determined by real time PCR and ELISA, respectively. Corneal sutures produced a consistent inflammatory response and a time-dependent neovascularization. The response in HO-2 null mice was associated with a greater increase compared to the wild type in the number of leukocytes (827,600+/-129,000 vs. 294,500+/-57,510; p<0.05), neovessels measured by vital microscopy (21.91+/-1.05 vs. 12.77+/-1.55 mm; p<0.001) 4 days after suture placement. Hemangiogenesis but not lymphangiogenesis was more pronounced in HO-2 null mice compared to wild type mice. Induction of HO-1 in sutured corneas was greatly attenuated in HO-2 null corneas and treatment with biliverdin diminished the exaggerated inflammatory and neovascular response in HO-2 null mice. The demonstration that the inflammatory responses, including expression of proinflammatory proteins, inflammatory cell influx and hemangiogenesis are exaggerated in HO-2 knockout mice strongly supports the notion that the HO system is critical for controlling the inflammatory and neovascular response in the cornea. Hence, pharmacological amplification of this system may constitute a novel therapeutic strategy for the treatment of corneal disorders associated with excessive inflammation and neovascularization.

Prevention of bacterial colonization of contact lenses with covalently attached selenium and effects on the rabbit cornea.

PURPOSE: Although silicone hydrogel materials have produced many corneal health benefits to patients wearing contact lenses, bacteria that cause acute red eye or corneal ulcers are still a concern. A coating that inhibits bacterial colonization while not adversely affecting the cornea should improve the safety of contact lens wear. A covalent selenium (Se) coating on contact lenses was evaluated for safety using rabbits and prevention of bacterial colonization of the contact lenses in vitro. METHODS: Contact lenses coated with Se were worn on an extended-wear schedule for up to 2 months by 10 New Zealand White rabbits. Corneal health was evaluated with slit-lamp biomicroscopy, pachymetry, electron microscopy, and histology. Lenses worn by the rabbits were analyzed for protein and lipid deposits. In addition, the ability of Se to block bacterial colonization was tested in vitro by incubating lenses in a Pseudomonas aeruginosa broth followed by scanning electron microscopy of the contact lens surface. RESULTS: The covalent Se coating decreased bacterial colonization in vitro while not adversely affecting the corneal health of rabbits in vivo. The Se coating produced no noticeable negative effects as observed with slit-lamp biomicroscopy, pachymetry, electron microscopy, and histology. The Se coating did not affect protein or lipid deposition on the contact lenses. CONCLUSION: The data from this pilot study suggest that a Se coating on contact lenses might reduce acute red eye and bacterial ulceration because of an inhibition of bacterial colonization. In addition, our safety tests suggest that this positive effect can be produced without an adverse effect on corneal health.

A comparison of gatifloxacin to ciprofloxacin in the prophylaxis of Streptococcus pneumoniae in rabbits in a LASIK model.

PURPOSE: To investigate the efficacy of the fourth-generation fluoroquinolone, gatifloxacin 0.3%, compared to ciprofloxacin 0.3%, in preventing Streptococcus pneumoniae keratitis in a rabbit laser in situ keratomileusis (LASIK) model. METHODS: Twelve albino rabbits had bilateral lamellar flaps created. Group A (eight eyes) was given gatifloxacin 0.3%; group B (eight eyes) was given ciprofloxacin 0.3%; and group C (eight eyes) served as the controls. Groups A and B received one drop of antibiotic 20 minutes before the creation of the lamellar flap, at the conclusion of flap formation, and four times per day for 3 days. All corneas were inoculated with 0.1 mL of 4 x 10 organisms/mL of S. pneumoniae immediately after flap formation. On day 3, all corneas were examined and cultured. RESULTS: Group A (gatifloxacin) had no infiltrates and three areas of 1-mm central corneal haze. On day 3, one of eight corneas had a positive culture. Group B (ciprofloxacin) had seven infiltrates, including one perforation, and six of eight corneas had positive cultures. Group C (control) had eight corneal infiltrates, and all eight corneas had positive cultures. The data show a statistically significant difference between gatifloxacin and ciprofloxacin and gatifloxacin and control for mean infiltrate size and mean culture scores. CONCLUSIONS: The fourth-generation fluoroquinolone, topical gatifloxacin 0.3%, is superior to topical ciprofloxacin 0.3% for prophylaxis against a clinical isolate of S. pneumoniae in a rabbit LASIK model.

Evaluation of preoperative and postoperative prophylactic regimens for prevention and treatment of diffuse lamellar keratitis.

PURPOSE: To investigate preoperative and postoperative prophylactic treatment with different pharmacological agents before flap cutting and exposure to a diffuse lamellar keratitis (DLK) causative agent. SETTING: Magill Research Center for Vision Correction, Storm Eye Institute, Medical University of South Carolina, Charleston, South Carolina, USA. METHODS: The study comprised 48 eyes of 24 Dutch-belted rabbits. Three days before a corneal flap was cut and the corneal interface was exposed to Pseudomonas aeruginosa lipopolysaccharide endotoxin, a DLK causative agent, the eyes were randomly assigned to treatment with a mast-cell stabilizer, a nonsteroidal antiinflammatory drug (NSAID), or a corticosteroid or left without treatment as controls. The treatment was maintained throughout the 1-week follow-up. Slitlamp examinations and photographs were performed at 1, 3, 5, and 7 days; DLK was graded by a masked observer from 0 (no DLK) to IV. Corneal interface scrapings were performed in selected eyes on day 7. RESULTS: At the end of the follow-up, 36 eyes were available for evaluation. At 1 week, 100% of the control eyes and the eyes treated with the mast-cell stabilizer developed DLK; in the NSAID-treated and corticosteroid-treated eyes, the DLK rate was 86% and 70%, respectively. At 1 day, the severity of DLK was significantly lower in eyes treated with the mast-cell stabilizer (0.44) and at 7 days, it was significantly lower in corticosteroid-treated eyes (0.3) than in the control group (1.5 and 1.4, respectively) (P<.05, Wilcoxon test). Corneal interface scraping from an eye with grade III DLK showed numerous inflammatory cells. CONCLUSIONS: Preoperative and postoperative treatment with corticosteroids significantly reduced the severity of DLK compared to the untreated control eyes in this animal model. Treatment with a mast-cell stabilizer and an NSAID had less effect on the postoperative course of DLK.

Reduction of capsaicin-induced ocular pain and neurogenic inflammation by calcium antagonists.

PURPOSE: To examine whether blockade of chemosensitivity of corneal nociceptors by Ca2+ antagonists decreases pain and irritation induced by capsaicin. METHODS: In adult rabbits, the number of lid-squeezing movements and the degree of palpebral opening, miotic response, and conjunctival vasodilation evoked by a bilateral instillation of 30 microliters of capsaicin (33 mM) were measured at different times (up to 5 hours) after the drug. Irritative responses to capsaicin in eyes pretreated with diltiazem, verapamil, or nifedipine were compared with those that received only the vehicle. Protein content in aqueous humor was also measured at the end of the experiment. RESULTS: Diltiazem at doses of 1 to 28 mM, administered 15 minutes before the application of capsaicin, significantly decreased scratching movements, conjunctival hyperemia, closure of the eye, and elevated aqueous protein concentration induced by capsaicin; however, it did not significantly reduce miosis. Nifedipine (2.8 and 10 mM) diminished the number of scratching movements but not other inflammatory parameters, whereas verapamil (2.8 and 10 mM) was totally ineffective in attenuating ocular signs of irritation produced by capsaicin. CONCLUSIONS: These results suggest that by lowering capsaicin-induced neural activity in nociceptive terminals, diltiazem decreases pain and neurogenic inflammation and may be useful as both an analgesic and an antiinflammatory agent in the eye.

[Causes and incidence of short-life keratitis caused by dacryography. Optimization of protective parameters]. / Cause e incidenze delle cheratopatie fugaci da dacriografia. Ottimizzazione dei parametri protettivi.

This study was aimed at investigating the changes occurring in superficial corneal epithelium (localized or diffuse, dotted or linear) as observed in 300 eyes after macrodacryography with iodate contrast media (iodized oil and water soluble non-ionic agents). In our opinion, the causes of iatrogenic short-life keratitis are: needle injury, the deposition of iodate contrast medium on the cornea and the reduction of palpebral winking, favoring dry eye, due to superficial anesthesia. The pharmacologic protection of the cornea by means of high-viscosity drugs allows both the number and the degree of keratitis to be markedly reduced. Therefore, contrast media must be chosen on the basis of anamnestic and clinical data, as well as of patient's symptoms, focusing mainly on the characteristics of the various agents--i.e., density, concentration, viscosity.

The use of prophylactic eye drops during high-dose cytosine arabinoside therapy.

An ocular toxic reaction presenting as conjunctivitis or keratitis develops in a significant number of patients who are treated with high-dose cytosine arabinoside (ara-C). Although eye drops containing glucocorticoid reportedly decrease the incidence, they do not totally eliminate this side effect. In comparing this technique with artificial tears, both were found to be equally effective. The primary mechanism by which eye drops decrease ocular toxic reactions associated with high-dose ara-C is presumably due to dilution of intraocular concentrations of ara-C.

The influence of a fish oil dietary supplement on immunogenic keratitis.

Fish lipids contain large amounts of eicosapentaenoic acid (EPA) and docosahexaenoic acid. These fatty acids are known to have an influence on prostaglandin (PG) and leukotriene (LT) synthesis. We studied the effect of a fish oil dietary supplement on an immune-complex-induced keratitis of the rabbit eye and compared it with the effect of a sunflower seed oil dietary supplement, rich in linoleic acid. Immune complex keratitis induced by intrastromal injection of human serum albumin (HSA) was characterized by leukocyte infiltrate, neovascularization, and corneal edema. Animals given a fish oil diet showed significantly less leukocyte infiltrate, neovascularization, and corneal edema, compared to those given a sunflower seed oil diet.

Inhibition by phosphoramidon of Pseudomonas aeruginosa elastase injected intracorneally in rabbit eyes.

Phosphoramidon (N-(alpha-L-rhamnopyranosyloxyhydroxyphosphinyl)-L-leucyl-L-trypto phan) is a powerful inhibitor of Pseudomonas aeruginosa elastase. The addition of this compound to an elastase solution injected intrastromally in rabbit eyes protected the corneas from the damage of the enzyme for a period of 12 hours. Phosphoramidon is comparable in this respect to the inhibitor 2-mercaptoacetyl-L-phenylalanyl-L-leucine, but is considerably more effective than its analog phosphoryl-L-leucyl-L-phenylalanine (P-Leu-Phe). It is suggested that the rhamnopyranosyl moiety, present in phosphoramidon but not in P-Leu-Phe, is responsible for the difference in the intracorneal activity of the two phosphoramidates. Phosphoramidon as well as the mercaptoacetyl derivative might prove beneficial in the treatment of Pseudomonas aeruginosa corneal infections.

Evaluation of BL-P1654: a semisynthetic penicillin as a topical ocular therapy.

BL-P1654 is a new semisynthetic penicillin that possesses broad spectrum antimicrobial activity against many gram-positive and gram-negative bacteria, including Pseudomonas aeruginosa. An in vitro profile of BL-P1654 was established against strains of Staphylococcus aureus, Streptococcus pneumoniae, and Pseudomonas aeruginosa, three bacteria frequently associated with infections of the eye. The effectiveness of BL-P1654 in preventing the development of experimentally-induced keratitis by each of these bacteria was determined. The results of these experiments show BL-P1654 to be more effective than gentamicin and support further evaluation of the semisynthetic penicillin for ophthalmic indications.

Effectiveness of neomycin and polymyxin ointments: prevention of Staphylococcus Aureus keratitis in rabbits.

Combinations of neomycin sulfate and polymyxin B sulfate are commonly used in ophthalmic ointments for the treatment or the prevention of bacterial keratoconjunctivitis. In this study we evaluated the effectiveness of various ointments containing these two antibiotics, alone and in combination, in preventing Staphylococcus aureus keratitis in rabbits. Rabbit eyes were infected by intracorneal inoculation, treated topically with ointment and graded by gross observation 24 hours after inoculation. Treatment with ointments containing neomycin alone offered significant protection against these corneal infections. The polymyxin B ointments, as well as the vehicle controls, were ineffective in preventing S aureus infections in the rabbit eyes. However, by far, the most effective ointment formulations tested were the combination ointments and specifically those containing 1.75-3.50 mg neomycin and 3,000-6,000 units polymycin B per gram of ointment.