PURPOSE: Chronic use of topical hypotensive agents induces several side effects caused by preservatives. The purpose of this study was to evaluate the effects of prostaglandin analogs with varying concentrations of benzalkonium chloride (BAC), preservative-free (PF), and alternative preservatives on mouse corneal tissue. METHODS: Thirty-five, 8- to 10-week-old female C57BL/6 mice (five mice for each group) were used for this study. To the control group, we applied normal saline, and to each drug-treated group we applied 0.02% BAC, bimatoprost 0.01% (with BAC 0.02%), latanoprost 0.005% (with BAC 0.02%), travoprost 0.004% (with 0.001% polyquad) or tafluprost 0.0015% with/without 0.001% BAC, once a day (9 p.m.) for 4 weeks. Corneal fluorescein staining was evaluated in all groups. After harvest, the corneal tissues were embedded in paraffin and then Hematoxylin-Eosin stain was performed for histopathological examination. Immunofluorescence staining was done against TNF-alpha, IL-6, HLA DR, pJNK, and pAkt. RESULTS: In corneal fluorescein staining, severe punctate epithelial keratitis was seen in the groups of 0.02% BAC, 0.02% BAC containing bimatoprost 0.01% and latanoprost 0.005%. The surface desquamation, irregular surface, loss of cell borders, anisocytosis and stromal shrinkage were observed in the groups of BAC-containing eye drops. Moreover, the groups treated with BAC-containing eye drops have high inflammatory markers, significantly decreased cell viability-related signal, pAkt, and higher apoptosis-inducing signal, pJNK, than the control group. On the other hand, travoprost 0.004% and PF tafluprost 0.0015% have less cellular morphologic changes, lower inflammation, and higher cellular viability than BAC-containing formulations. CONCLUSIONS: Corneal damage, increased inflammation and apoptosis and low cell viability were observed in BAC-containing groups. PF or alternatively preserved glaucoma medications seem to be a reasonable and viable alternative to those preserved with BAC.
Animals ; Cell Survival ; Conjunctiva/drug effects/*pathology ; Disease Models, Animal ; Epithelium, Corneal/drug effects/*pathology ; Female ; Glaucoma/*drug therapy/pathology ; Mice ; Mice, Inbred C57BL ; Microscopy, Fluorescence ; Ophthalmic Solutions ; Preservatives, Pharmaceutical ; Prostaglandins, Synthetic/*administration & dosage

PURPOSE: To investigate the therapeutic effects of mineral oil (MO) and hyaluronic acid (HA) mixture eye drops on the tear film and ocular surface in a mouse model of experimental dry eye (EDE). METHODS: Eye drops consisting of 0.1% HA alone or mixed with 0.1%, 0.5%, or 5.0% MO were applied to desiccating stress-induced murine dry eyes. Tear volume, corneal irregularity score, tear film break-up time (TBUT), and corneal fluorescein staining scores were measured at 5 and 10 days after treatment. Ten days after treatment, goblet cells in the conjunctiva were counted after Periodic acid-Schiff staining. RESULTS: There was no significant difference in the tear volume between desiccating stress-induced groups. The corneal irregularity score was lower in the 0.5% MO group compared with the EDE and HA groups. The 0.5% and 5.0% MO groups showed a significant improvement in TBUT compared with the EDE group. Mice treated with 0.1% and 0.5% MO mixture eye drops showed a significant improvement in fluorescein staining scores compared with the EDE group and the HA group. The conjunctival goblet cell count was higher in the 0.5% MO group compared with the EDE group and HA group. CONCLUSIONS: The MO and HA mixture eye drops had a beneficial effect on the tear films and ocular surface of murine dry eye. The application of 0.5% MO and 0.1% HA mixture eye drops could improve corneal irregularity, the corneal fluorescein staining score, and conjunctival goblet cell count compared with 0.1% HA eye drops in the treatment of EDE.
Animals ; Conjunctiva/*drug effects/pathology ; Cornea/metabolism ; Disease Models, Animal ; Drug Combinations ; Dry Eye Syndromes/*drug therapy/metabolism ; Emollients/administration & dosage ; Female ; Goblet Cells/drug effects/metabolism/pathology ; Hyaluronic Acid/*administration & dosage ; Mice ; Mice, Inbred C57BL ; Mineral Oil/*administration & dosage ; Ophthalmic Solutions ; Tears/*metabolism ; Viscosupplements/administration & dosage

Vascular endothelial growth factor inhibitor is an emerging therapeutic modality for various ocular diseases with neovascularization (NV). However, for corneal NV, controversy remains regarding whether bevacizumab or ranibizumab is superior. A 32-year-old female diagnosed with herpetic keratoconjunctivitis with refractory corneal NV despite two previous subconjunctival and intrastromal bevacizumab injections, received two subconjunctival and intrastromal ranibizumab injections. Six months postoperatively, there was significant regression of the neovascular area and vessel caliber. Here, the authors report a case of improvement in corneal NV with subconjunctival and intrastromal ranibizumab injections, which was previously refractory to bevacizumab injection. The findings may suggest a new prospect in treating corneal NV.
Adult ; Angiogenesis Inhibitors/administration & dosage ; Antibodies, Monoclonal, Humanized/*administration & dosage ; Conjunctiva/blood supply ; Corneal Neovascularization/*drug therapy ; Corneal Stroma/blood supply ; Female ; Humans ; Injections, Intraocular/methods ; Keratitis, Herpetic/*drug therapy ; Visual Acuity/drug effects

Adult ; Anti-Bacterial Agents ; therapeutic use ; Conjunctiva ; drug effects ; microbiology ; Female ; Humans ; Keratomileusis, Laser In Situ ; adverse effects ; methods ; Levofloxacin ; therapeutic use ; Male ; Povidone-Iodine ; therapeutic use ; Prospective Studies

PURPOSE: Long-term use of topical medication is needed for glaucoma treatment. One of the most commonly prescribed classes of hypotensive agents are prostaglandin analogs (PGs) used as both first-line monotherapy; as well as in combination therapy with other hypotensive agents. Several side effects of eye drops can be caused by preservatives. The purpose of this study was to evaluate the effects of PGs with varying concentrations of benzalkonium chloride (BAC), alternative preservatives, or no preservatives on human conjunctival fibroblast cells. METHODS: Primary human conjunctival fibroblast cells were used in these experiments. Cells were exposed to the following drugs: BAC at different concentrations, bimatoprost 0.01% (with BAC 0.02%), latanoprost 0.005% (with BAC 0.02%), tafluprost 0.0015% with/without 0.001% BAC and travoprost 0.004% (with 0.001% Polyquad) for 15 and 30 minutes. Cell cytotoxicity was evaluated by phase-contrast microscopy to monitor morphological changes of cells, Counting Kit-8 (CCK-8) assay to cell viability, and fluorescent activated cell sorting (FACS) analysis to measure apoptosis. RESULTS: BAC caused cell shrinkage and detachment from the plate in a dose-dependent manner. Morphological changes were observed in cells treated with bimatoprost 0.01% and latanoprost 0.005%. However, mild cell shrinkage was noted in cells treated with tafluprost 0.0015%, while a non-toxic effect was noted with travoprost 0.004% and preservative-free tafluprost 0.0015%. CCK-8 assay and FACS analysis showed all groups had a significantly decreased cell viability and higher apoptosis rate compared with the control group. However, travoprost 0.004% and preservative-free tafluprost 0.0015% showed lower cytotoxicity and apoptosis rate than other drugs. CONCLUSIONS: This in vitro study revealed that BAC-induced cytotoxicity is dose-dependent, although it is important to emphasize that the clinical significance of toxicity differences observed among the different PGs formulations has not yet been firmly established. Alternatively preserved or preservative-free glaucoma medications seem to be a reasonable and viable alternative to those preserved with BAC.
Apoptosis ; Cell Line ; Cell Survival/drug effects ; Conjunctiva/drug effects/*pathology ; Fibroblasts/drug effects/pathology ; Glaucoma/drug therapy/pathology ; Humans ; Preservatives, Pharmaceutical/*pharmacology ; Prostaglandins, Synthetic/*pharmacology

PURPOSE: The purpose of this study was to investigate the myotoxicity of bevacizumab on extraocular muscles in a rabbit model. METHODS: Thirty New Zealand white rabbits were used for this study. The animals were evenly divided into two groups. In the first group, 15 rabbits were treated with intramuscular injections of bevacizumab (1.25 mg/0.05 mL) in the right superior rectus muscle and normal saline solution (0.05 mL) in the left superior rectus muscle. In the second group, 15 rabbits were treated with subconjunctival injections of bevacizumab (2.5 mg/0.1 mL) in the right superior subconjunctival area and normal saline solution (0.1 mL) in the left superior subconjunctival area. Five rabbits in each group were sacrificed at one day, two weeks and four weeks after the injections. Extraocular muscle samples were prepared for light microscopic (LM) and electron microscopic (EM) examination. Degrees of acute inflammation were evaluated via CD-11b immunohistochemistry, and global muscle change was investigated using hematoxylin and eosin stains. Intensity of fibrosis was evaluated using Masson trichrome stains, and ultrastructural changes were observed on EM. RESULTS: We observed no significant inflammatory cell infiltration, muscle necrosis or fibrotic change in treated and control eyes. EM findings revealed no significant damage to muscle or vascular tissue after bevacizumab injection. CONCLUSIONS: We found no signs of extraocular muscle toxicity after LM and EM intramuscular and subconjunctival bevacizumab injections in a rabbit model.
Angiogenesis Inhibitors/*administration & dosage/toxicity ; Animals ; Antibodies, Monoclonal, Humanized/*administration & dosage/toxicity ; Conjunctiva/drug effects ; Injections ; Oculomotor Muscles/*drug effects ; Rabbits

This prospective observational case series study included 6 eyes of 6 consecutive glaucomatous patients. Each patient underwent trabeculectomy with mitomycin C, and received a 1.25 mg of subconjunctival bevacizumab injection at completion of the trabeculectomy. Study eyes included two with neovascular glaucoma, three with uveitic glaucoma, and one with secondary glaucoma following vitrectomy. All eyes had undergone failed glaucoma laser/surgical treatment or an intraocular surgical procedure. Intraocular pressure (IOP) at the following postoperative visits: preoperative, 1 week, 1 month, 2 months, 3 months, and 6 months, was measured. We also evaluated postoperative bleb findings and complications. IOP measured at each visit was 37.5+/-14.4 mmHg, 6.2+/-3.4 mmHg, 8.3+/-7.2 mmHg, 12.0+/-4.4 mmHg, 10.8+/-3.1 mmHg, and 12.2+/-3.3 mmHg, respectively, for each visit. All eyes had functioning blebs with normal IOP at postoperative 6 months with no additional IOP-lowering medication.
Adult ; Aged ; Angiogenesis Inhibitors/*administration & dosage ; Antibodies, Monoclonal/*administration & dosage ; Conjunctiva ; Female ; Glaucoma/*drug therapy/etiology/*surgery ; Glaucoma, Neovascular/drug therapy/surgery ; Humans ; Injections, Intraocular ; Male ; Middle Aged ; Prospective Studies ; Trabeculectomy/*methods ; Uveitis/complications ; Vascular Endothelial Growth Factor A/*antagonists & inhibitors ; Vitrectomy/adverse effects

PURPOSE: To investigate the short-term efficacy of topical immunosuppressive agents on the survival of cultivated allo-conjunctival equivalents. METHODS: Twenty-five eyes of New Zealand white rabbits were included. Temporal conjunctivae were trephined to a diameter of 7.5 mm, and then cultured allo-conjunctival epithelial cells on amniotic membrane were transplanted onto them. Various immunosuppressants including steroid, cyclosporine, and rapamycin were applied topically four times a day for a week. Epithelial defects and graft edema were graded daily. Numbers of inflammatory cells were measured in H&E. PKH26 and cytokeratin 4 and 7 were immunostained. RESULTS: Earlier epithelialization was observed in 1% steroid-treated eyes and defects persisted significantly in 0.5% CsA applied eyes. In histology, PKH26 positive cells considered as donor cells were only found in 1% steroid or 0.01% rapamycin applied eyes. 1% steroid- or 0.01% rapamycin-applied eyes both showed positive staining for keratin-4 and -7. Inflammatory cells were less found in 1% steroid or 0.01% rapamycin treated eyes. CONCLUSIONS: Topical steroid or rapamycin can help to suppress acute inflammation and enhance the acute survival of transplanted conjunctival cells.
Administration, Topical ; Animals ; Cell Count ; *Cell Transplantation ; Cells, Cultured ; Conjunctiva/*cytology ; Cyclosporine/pharmacology ; Epithelial Cells/metabolism/*transplantation ; Female ; Fluorescent Antibody Technique, Indirect ; Graft Survival/*drug effects ; Immunosuppressive Agents/*pharmacology ; Keratin-4/metabolism ; Keratin-7/metabolism ; Male ; Organic Chemicals/metabolism ; Prednisone/pharmacology ; Rabbits ; Sirolimus/pharmacology ; Transplantation, Homologous

PURPOSE: To compare the short term effects of topical 0.05% cyclosporine (CsA) and a mixture of 0.08% chondroitin sulfate and 0.06% sodium hyaluronate (CS-HA) on dry eye ocular surfaces. METHODS: 36 patients with moderate to severe dry eye (5 mm/5 min or less with Schirmer's test or tear break up time (BUT) less than 6 seconds), were treated with topical application of CS-HA on one eye and CsA on the other 4 times a day for 6-8 weeks. BUT, Schirmer's test without anesthesia, and conjunctival impression cytology (CIC; goblet cell density, nucleus to cytoplasmic ratio, and epithelial cell morphology) were evaluated and compared between eyes before and after treatment (repeated measurement of ANOVA). RESULTS: After treatment, BUT and tear wettings were significantly prolonged in each group. Topical CsA treated eyes had greater increase in BUT (p=0.026); there was no significant difference in tear wetting (p=0.132). While the 3 parameters of CIC improved in both groups, goblet cell density was significantly higher in eyes treated with CsA (p=0.033). CONCLUSIONS: While both CS-HA and 0.05% CsA eyedrops improve ocular surfaces, topical CsA may have a better effect on enhancing tear film stability and goblet cell density.
Adjuvants, Immunologic/administration & dosage ; Administration, Topical ; Cell Count ; Chondroitin Sulfates/*administration & dosage ; Conjunctiva/drug effects/pathology ; Cyclosporine/*administration & dosage ; Drug Administration Schedule ; Drug Combinations ; Drug Therapy, Combination ; Dry Eye Syndromes/*drug therapy/metabolism/pathology ; Epithelium/drug effects/pathology ; Female ; Follow-Up Studies ; Goblet Cells/drug effects/pathology ; Humans ; Hyaluronic Acid/*administration & dosage ; Immunosuppressive Agents/*administration & dosage ; Male ; Middle Aged ; Ophthalmic Solutions/administration & dosage ; Tears/drug effects/metabolism ; Time Factors ; Treatment Outcome

OBJECTIVEElucidating the detoxification mechanism of the raw Pinellia ternata processed by alum solution or alkaline solution (pH > 12).

METHODRaw Pinellia ternata was immersed in alum solution and alkaline solution according to Chinese pharmacopoeia. Observed the shape's changing of needle-like calcium oxalate crystals by scanning electro-microscopy. Determinating the contents of calcium oxalate crystals by applying oxidation reduction titration. Measured the irritations of raw P. ternata and various processing products on the model of rabbits'eyes.

RESULTAfter processed by 8% alum solution prescribed in Chinese pharmacopoeia or 10% sodium carbonate solution, the needle-like shape of raphides in raw Pinellia ternata were changing and the sting barb of raphides were rusted and dissolved, the contents of calcium oxalate crystal in raw Pinellia ternata were sharply declined from more than 1% to less than 0.5%. the decline of contents is relative to the irritation decline of P. ternata on rabbit's eyes. Less than 0.5% calcium oxalate crystals of P. ternata almost had no irritation.

CONCLUSIONAfter processed by 8% alum solution or sodium carbonate solution (pH > 12) , the irritation components in raw P. ternata could be rusted and dissolved, the needle point of raphides was broken, which led to the raphides'content declining and the irritation disappearing. The micro-structures, shapes and contents of calcium oxalate crystals in different medicine plants were not same. These properties of calcium oxalate crystal in India Madder Root and yam et al were very different from those in raw P. ternata.

Alum Compounds ; chemistry ; Animals ; Calcium Oxalate ; analysis ; isolation & purification ; toxicity ; Carbonates ; chemistry ; Conjunctiva ; drug effects ; Crystallization ; Drug Interactions ; Eye Diseases ; chemically induced ; Pinellia ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Powders ; Rabbits ; Technology, Pharmaceutical ; methods