Clinical efficacy of a new surgical technique of oral mucosal epithelial transplantation for severe ocular surface disorders.

BACKGROUND: Severe ocular surface disorders are one of the major blinding diseases, and a paucity of original tissue obscures successful reconstruction. We developed a new surgical technique of direct oral mucosal epithelial transplantation (OMET) to reconstruct severely damaged ocular surfaces in 2011. This study elaborates on the clinical efficacy of OMET. METHODS: A retrospective review of patients with severe ocular surface disorders who underwent OMET from 2011 to 2021 at the Department of Ophthalmology, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine was conducted. Patients who were followed up for at least 3 months postoperatively and had sufficient pre or postoperative records were included. Surgical efficacy was evaluated by comparing the best-corrected visual acuity (BCVA), corneal transparency, neovascularization grade, and symblepharon grade. Additionally, postoperative ocular surface impression cytology was used to study the morphology of the newborn epithelial cells. RESULTS: Forty-eight patients (49 eyes; mean age: 42.55 ± 12.40 years, range:12-66 years) were enrolled in the study. The etiology included chemical burns (30 eyes), thermal burns (16 eyes), explosive injuries (1 eye), Stevens-Johnson syndrome (1 eye), and multiple pterygiums (1 eye). The mean follow-up period was 25.97 ± 22.99 months. Postoperatively, 29 eyes (59.18%) showed improved corneal transparency, 26 eyes (53.06%) had improved BCVA, 47 eyes (95.92%) had a stable epithelium until the final follow-up, 44 eyes (89.80%) had a reduced neovascularization grade. Of the 20 eyes with preoperative symblepharon, 15 (75%) were completely resolved, and five (25%) were partially resolved. Impression cytological studies showed no postoperative conjunctival invasion onto the corneal surface. CONCLUSIONS: OMET is a safe and effective surgical technique for reconstruction in severe ocular surface disorder by maintaining a stable epithelium and reducing the neovascularization and symblepharon grade.

Corneal endothelial regeneration in human eyes using endothelium-free grafts.

BACKGROUND: To report on corneal endothelial regeneration, graft clarity, and vision recovery when using endothelium-free grafts. METHODS: We evaluated the donor's cell viability using trypan blue staining and dual staining with calcein acetoxy methyl ester and ethidium homodimer-1. To preserve eyeball integrity, we performed therapeutic penetrating keratoplasty using cryopreserved donor tissue without endothelium on 195 consecutive patients who suffered from corneal perforation due to progressive primary corneal disease such as herpes simplex keratitis, fungal keratitis, ocular thermal burns, keratoconus, and phlyctenular keratoconjunctivitis. Of these, 18 eyes recovered corneal graft clarity and underwent periodic slit-lamp microscopy, A-scan pachymetry, and in vivo confocal microscopy to observe the clinical manifestations, variations in corneal thickness, and repopulation of the corneal endothelial cells on the donor grafts. RESULTS: No viable cells were detected in the cryopreserved corneas. After the therapeutic penetrating keratoplasty, notable corneal graft edema was observed in all 18 eyes for 1-4 months, and no corneal endothelial cells were detected on the grafts during this period. Thereafter, we observed gradual and progressive regression and final resolution of the stromal edema, with complete recovery of corneal graft clarity. Through periodic confocal microscopy, we observed the corneal endothelium's regenerating process, along with single cells bearing multiple nuclei and cell division-like morphology. The regenerated endothelium on the grafts reached a mean cell density of 991 cells/mm2. Remarkable vision rehabilitation was achieved in all 18 patients. CONCLUSIONS: We obtained conclusive evidence that host-derived endothelial cells can regenerate a new endothelium over the endothelium-free graft, which possesses normal functions for corneal clarity and vision recovery.

Anti-fibrosis potential of pirarubicin via inducing apoptotic and autophagic cell death in rabbit conjunctiva.

This study investigated the potential efficacy of pirarubicin (THP) in modulating rabbit conjunctival fibrosis both in vitro and in vivo and characterized the underlying mechanisms. Primary rabbit conjunctival fibroblasts (RCF) were cultured and treated with THP or mitomycin C (MMC) for 5 min, followed by assaying for cell viability, cell cycle distribution, apoptotic and autophagic pathways. The production of reactive oxygen species (ROS) and chemotaxis of macrophages by RCF were evaluated using 2',7'-dichlorofluorescein diacetate (DCFH-DA) labeling and transwell migration assay, respectively. Limbal stem cell excision in combination with alkali burn was performed on the rabbits to establish a model of limbal deficiency and conjunctival fibro-vascular invasion. After three months, the modeled fibro-vascular tissue was excised combined with topical subconjunctival 5-min exposure to THP compared with MMC intraoperatively. The recurrence of postoperative fibrosis and the expression of apoptosis, autophagy, and inflammation markers were evaluated by immunohistochemistry. All modeled rabbits developed conjunctival fibro-vascular lesions, which were similar to human recurrent pterygium (HRP). Both THP and MMC inhibited RCF proliferation and arrested cell cycle at the G0/G1 phase. In particular, 7.5 µmol/L THP remarkably promoted RCF autophagy by upregulating the levels of Beclin 1, Atg 5/12 conjugate, and LC3B, whereas, 15 µmol/L THP significantly triggered a cascade of mitochondrial-associated RCF apoptosis. THP induced the production of ROS and enhanced the chemoattraction of macrophages by RCF. Similar to 600 µmol/L MMC, both 7.5 µmol/L and 15 µmol/L THP attenuated postoperative conjunctival fibrosis in the models; 7.5 µmol/L THP preferentially enhanced autophagy while causing fewer side effects. THP exerted its antifibrotic action by modulating autophagy in RCF, inducing cell cycle arrest, and mitochondrial-mediated apoptosis. THP at the dose of 7.5 µmol/L prevented postoperative conjunctival fibrosis in an animal model.

New histopathologic and ultrastructural findings in Reis-Bücklers corneal dystrophy caused by the Arg124Leu mutation of TGFBI gene.

BACKGROUND: Reis-Bücklers corneal dystrophy (RBCD) was consistently reported as a corneal dystrophy only affected Bowman's layer and superficial corneal stroma, and superficial keratectomy was a recommendation surgery for treatment in literatures. The study reported new histopathological and ultrastructural findings in RBCD caused by the Arg124Leu mutation of transforming growth factor induced (TGFBI) gene in a four-generation Chinese pedigree. METHODS: Subjects including eight patients and seven unaffected family members received slit-lamp biomicroscopy and photography. DNA was obtained from all subjects, and exons 4 and 11 to 14 of TGFBI gene were analyzed by polymerase chain reaction and the products were sequenced. Anterior segment optical coherence tomography (AS OCT) and in vivo confocal microscopy were conducted for ten eyes of five patients. Based on the results of AS OCT and in vivo confocal microscopy, deep anterior lamellar keratoplasty (DLKP) using cryopreserved donor cornea was applied for four eyes of four patients. Four lamellar dystrophic corneal buttons were studied by light and transmission electron microscopy, and TGFBI immunohistochemistry. RESULTS: Eight patients had typical clinical manifestations of RBCD presenting recurrent painful corneal erosion starting in their early first decades, along with age-dependent progressive geographic corneal opacities. TGFBI sequencing revealed a heterozygous mutation, Arg124Leu in all eight patients. Anterior segment optical coherence tomography and in vivo confocal microscopy showed the dystrophic deposits involved not only in subepithelial and superficial stroma, but also in mid- or posterior stroma in four examined advanced eyes. Light microscopy showed Bowman's layer was absent, replaced by abnormal deposits stain bright red with Masson's trichrome. In superficial cornea, the deposits stacked and produced three to five continuous bands parallel to the corneal collagen lamellae. In mid- to posterior stroma, numerous granular or dot- like aggregates were heavily scattered, and most of them presented around the nuclei of stromal keratocytes. Transmission electron microscopy revealed the multiple electron-dense rod-shaped deposits aggregated and formed a characteristic pattern of three to five continuous bands in superficial cornea, which were similar to those seen under light microscopy. In mid- to posterior stroma, clusters of rod-shaped bodies were scattered extracellular or intracellular of the stromal keratocytes between the stromal lamellae suggesting the close relationship between mutated proteins and keratocyte. CONCLUSIONS: The study offer evidences indicating DLKP is a viable treatment option for advanced RBCD to avoid recurrence, and the mutated TGFBIp in dystrophic corneas are of keratocytes origin.

The role of ultraviolet radiation in the pathogenesis of pterygia (Review).

Pterygium is a common ophthalmic disease affecting humans only. Extensive epidemiological data have demonstrated a causative effect of chronic ultraviolet (UV) radiation on pterygia. Progress has been made in determining the origin of pterygia, their nasal predilection and wing­shaped appearance, and the roles of UV radiation in the initiation and the development of pterygia. In the present review, the current understanding of the involvement of UV radiation in the pathogenesis of pterygia is summarized. This involvement includes the alteration of limbal stem cells and fibroblasts that contribute to the initiation of pterygia and the induction of various pro­inflammatory cytokines, growth factors and matrix metalloproteinases that promote the progression of pterygia. Further elucidation of the roles of UV radiation in the pathogenesis of pterygia may help to encourage individuals at risk of developing pterygia to take preventive measures and aid researchers in the development of novel targeted therapeutic agents to treat pterygia.

Peripheral deep anterior lamellar keratoplasty using a cryopreserved donor cornea for Terrien's marginal degeneration.

OBJECTIVE: To evaluate the clinical efficacy of peripheral deep anterior lamellar keratoplasty (DALK) using a cryopreserved donor cornea for Terrien's marginal degeneration (TMD). METHODS: Thirty-one eyes of 27 patients with TMD underwent peripheral DALK using cryopreserved donor corneas. According to the distance between the inner edge of the lesion and the limbus, a ring-shaped or D-shaped DALK was performed. All grafts were stored at -20 °C. Cryopreserved corneoscleral rims were prepared for ring-shaped grafts and cryopreserved whole eyeballs were prepared for D-shaped grafts. The general conditions, intraoperative performance, postoperative corneal reconstruction, astigmatism, best corrected visual acuity (BCVA), and various complications were analyzed. RESULTS: Ring-shaped DALK was performed in 28 eyes and D-shaped DALK was performed in 3 eyes. Postoperative follow-up time was (28.4±24.8) months. There was evidence of inflammation before surgery in 12 eyes (38.7%) and intraoperative perforation occurred in 13 eyes (41.9%). The corneal structures of all eyes were reconstructed. Postoperative astigmatism and BCVA showed improvement (both P=0.00) except for cases that underwent D-shaped DALK. Ten eyes (32.2%) developed transient ocular hypertension and one eye (3.2%) developed secondary glaucoma. No primary disease recurrence or corneal allograft rejection was observed. CONCLUSIONS: Peripheral DALK for TMD using cryopreserved donor tissue is an effective technique that eliminates rejection and extends the use of donor eyes. Inflammatory history or intraoperative perforation has no adverse effect on graft recovery. However, D-shaped DALK did not achieve good visual outcomes.

Mycotic keratitis caused by concurrent infections of Exserohilum mcginnisii and Candida parapsilosis.

BACKGROUND: Mycotic keratitis in human cornea has been rarely reported to be associated with a co-infection of filamentous fungi and yeast. This paper aims to report a case of mycotic keratitis concurrently infected by Exserohilum mcginnisii and Candida parapsilosis. CASE PRESENTATION: A Chinese female presented two superposed corneal infiltrates with different size and texture on her left eye. In vivo confocal microscopy showed hyper-reflective multiple linear with highly branching structures distributing in the anterior corneal stroma. Inoculations of the corneal lesion scrape concurrently grew two similar superposed colonies on Sabouraud dextrose and chocolate agar plate. The larger colony exhibited mould, cottony and floccose at the edge, while the smaller one showed creamy and shiny surface. Modified slide culture for mould revealed hyphae were septate, and conidia were brown, smooth-walled, cylindrical to slight clavate with 6 to 13 pseudosepta. Based on the morphology of microscopic and macroscopic characteristics, the mould was identified as Exserohilum mcginnisii. Smear of the non-mould colony showed ellipse or ovoid budding yeast-like cells with abundant pseudomycelium. Vitek Yeast Biochemical Card test identified the yeast as Candida parapsilosis. With treatment of combined oral itraconazole with topical amphotericin B, a complete resolution of the corneal infiltrate was achieved within 1.5 months. CONCLUSION: This is the first documented case of human corneal infection by Exserohilum mcginnisii, and also the first report providing evidence of mycotic keratitis in human cornea concurrently infected by filamentous fungi and yeast.

Long-term comparison of full-bed deep lamellar keratoplasty with penetrating keratoplasty in treating corneal leucoma caused by herpes simplex keratitis.

PURPOSE: To compare long-term outcomes of full-bed deep lamellar keratoplasty (DLK) with penetrating keratoplasty (PK) for treating corneal leucoma caused by herpes simplex keratitis (HSK). DESIGN: Retrospective, comparative, interventional case series. METHODS: setting: Institutional. patients: Inclusion criteria were patients with corneal scarring induced exclusively by HSK who underwent primary graft of full-bed DLK or PK and completed a minimum of 12 months postoperative follow-up. There was no significant difference of corneal scarring and vascularization between the 2 groups before surgery. Choosing PK or full-bed DLK depended on the patient's own willingness, except those patients with a preoperative endothelial cell count of less than 700 cells/mm(2) or whose endothelial cell count was undetectable were encouraged to undergo only PK. Exclusion criteria were patients with a past history of corneal perforation, nonprimary graft, non-HSK-related corneal scars, and failure to complete a minimum of 12 months of postoperative follow-up. Fifty-eight eyes of 58 patients in the full-bed DLK group and 63 eyes of 63 patients in the PK group met the inclusion criteria. main outcome measures: Postoperative managements, recurrence of HSK, graft rejection, graft survival rate, visual acuity, and corneal endothelial density. RESULTS: The mean postoperative follow-up duration was 45.8 ± 30.9 months in the full-bed DLK group and 47.9 ± 27.2 months in the PK group (P = .70). As compared with the PK group, the full-bed DLK group experienced earlier suture removal (P = .01), needed fewer postoperative visits (P < .001), and had a higher proportion of eyes with full withdrawal of oral acyclovir (P < .001) and topical corticosteroid (P < .001). There were a total of 21 episodes of recurrent HSK in the PK group, more frequent than the 7 episodes in the full-bed DLK group, among which recurrent epithelial keratitis amounted to 13 episodes in the PK group, remarkably more frequent than the 1 episode in the full-bed DLK group. Twenty-six eyes (41.3%) encountered rejection episodes in the PK group, but no rejection episode was found in the full-bed DLK group (P < .001). In 14 eyes in the PK group, graft failure developed because of graft rejection, recurrence of HSK, or both, whereas only in 1 eye in the full-bed DLK group did graft failure develop because of recurrence of HSK (P = .001). The clear graft survival rate in the full-bed DLK group was significantly higher than that in the PK group (P = .01). Corneal endothelial cell density was stable from 1 month through 5 years in the full-bed DLK group, but 51.3% cell loss was found in the PK group at 5 years after surgery. At the last visit, 66.1% of eyes with full-bed DLK grafts and 50.9% of eyes with PK grafts achieved a best-correct visual acuity of 0.5 or better (P = .10). CONCLUSIONS: Advantages of full-bed DLK over PK are no allograft rejection, longer graft survival, earlier drug withdrawal of topical steroid and oral acyclovir, less recurrence of HSK, and fewer follow-up visits. Full-bed DLK is preferable for treating HSK-induced corneal scarring with relatively healthy endothelium and with no history of perforation.

Roles of the 15-kDa selenoprotein (Sep15) in redox homeostasis and cataract development revealed by the analysis of Sep 15 knockout mice.

The 15-kDa selenoprotein (Sep15) is a thioredoxin-like, endoplasmic reticulum-resident protein involved in the quality control of glycoprotein folding through its interaction with UDP-glucose:glycoprotein glucosyltransferase. Expression of Sep15 is regulated by dietary selenium and the unfolded protein response, but its specific function is not known. In this study, we developed and characterized Sep15 KO mice by targeted removal of exon 2 of the Sep15 gene coding for the cysteine-rich UDP-glucose:glycoprotein glucosyltransferase-binding domain. These KO mice synthesized a mutant mRNA, but the shortened protein product could be detected neither in tissues nor in Sep15 KO embryonic fibroblasts. Sep15 KO mice were viable and fertile, showed normal brain morphology, and did not activate endoplasmic reticulum stress pathways. However, parameters of oxidative stress were elevated in the livers of these mice. We found that Sep15 mRNA was enriched during lens development. Further phenotypic characterization of Sep15 KO mice revealed a prominent nuclear cataract that developed at an early age. These cataracts did not appear to be associated with severe oxidative stress or glucose dysregulation. We suggest that the cataracts resulted from an improper folding status of lens proteins caused by Sep15 deficiency.

Low levels of hydrogen peroxide stimulate corneal epithelial cell adhesion, migration, and wound healing.

PURPOSE: Intracellular reactive oxygen species have been reported to associate with growth factor and integrin signalings in promoting cell adhesion in many cell types. This study is to explore if exogenous H(2)O(2) at low levels can be beneficial to cell adhesion, migration, and wound healing. METHODS: Primary rabbit corneal epithelial cells treated with 0-70 µM H(2)O(2) were tested for viability by MTT assay, adhesion by centrifugation assay, focal contacts of vinculin and F-actin by immunofluorescence, activated Src(pY416), EGF receptor (pY845), vinculin(pY1065), FAK(pY397), and FAK(pY576) by immunoblotting. Cell migration was examined with 0-50 µM H(2)O(2) using the scratch wound technique. Corneal wound healing of ex vivo pig model and in vivo mouse model was examined using H(2)O(2) with and without antioxidant N-acetylcysteine (NAC). RESULTS: Compared with the untreated control, H(2)O(2) at 10-50 µM stimulated cell viability and facilitated adhesion and migration with clear induction of vinculin-rich focal adhesions and F-actin-containing stress fibers by increasing activated Src, FAK(pY576), and vinculin(pY1065). H(2)O(2) also increased phosphorylation of EGFR(Y845) parallel to that of activated Src, but both were eliminated by NAC and PP1 (Src inhibitor). Finally, H(2)O(2) induced faster wound healing in cornea both in vitro and in vivo, but the healing was diminished by NAC. CONCLUSIONS: These findings suggest that H(2)O(2) at low levels promotes cell adhesion, migration, and wound healing in cornea cells or tissue, and the interaction of H(2)O(2) with Src plays a major role.

Reactive oxygen species (ROS) are essential mediators in epidermal growth factor (EGF)-stimulated corneal epithelial cell proliferation, adhesion, migration, and wound healing.

EGF is an essential growth factor needed for epithelial cell proliferation and wound healing of the cornea, but the molecular mechanism is not understood. Although studies have shown that EGF in some non-phagocytic cells induces ROS generation, little is known about the role of ROS in corneal epithelial cells. Therefore, we examined the potential physiological role of ROS in corneal cell proliferation, adhesion and wound healing using rabbit or human corneal epithelial cells, and pig whole cornea organ culture as models. EGF (5 ng/ml)-induced ROS in serum-starved RCE or HCE cells were captured as DCFH fluorescence and detected by confocal microscopy. The elevation of ROS was eradicated when the cells were pretreated with an antioxidant N-acetylcysteine (NAC) or mannitol, or with inhibitor to NADPH oxidase (DPI), or to lipoxygenase (NDGA). EGF-induced ROS generation correlated with cell growth and activation of Akt and MAPK signaling pathways, while NAC eliminated all these effects. EGF-stimulated cell adhesion or migration in cell culture was greatly suppressed in the presence of NAC while EGF-facilitated epithelial cell wound healing in corneal organ culture was also blocked by NAC. This is the first demonstration of a novel ROS physiological function in corneal wound healing.

Recurrence of lattice corneal dystrophy caused by incomplete removal of stroma after deep lamellar keratoplasty.

PURPOSE: To report clinical and histopathological characteristics of postoperative amyloidosis recurrence in a patient with lattice corneal dystrophy (LCD) type I. METHODS: The clinical manifestation of recurrent amyloidosis in the residual stroma was delineated in a patient with LCD type I after deep lamellar keratoplasty (DLKP) for 6.5 years. Complete removal of the residual recipient stroma and regrafting of a new cryopreserved donor button were accomplished by a secondary DLKP. The primary DLKP donor graft and the underlying residual stroma of the recipient obtained by the secondary DLKP were examined for analysis of histopathologic and ultrastructural changes. RESULTS: A tongue-shaped retained stroma with linear opacity was observed underneath the primary DLKP donor graft. The retained stromal layer was thoroughly detached from Descemet membrane, removed, and followed by grafting a new cryopreserved button. The primary donor button exhibited a normal epithelium, fewer keratocytes, an intact Descemet membrane, and mild positive Congo red staining in the middle layer of the stroma. The total retained recipient stroma removed by the secondary DLKP measured approximately 20 mum in thickness, showing thick and massive amyloid accumulation. The surface of the removed residual stroma toward Descemet membrane showed collagen fibers in an interwoven fashion without bundle structure under a scanning electron microscope. CONCLUSION: Incomplete removal of the recipient stroma by DLKP can lead to the recurrence of amyloidosis in the residual stroma in patients with LCD. Clinical and histologic findings in the primary graft and in the residual recipient stroma implicate stromal genesis of recurrence of LCD after DLKP.

Mitomycin C, amniotic membrane transplantation and limbal conjunctival autograft for treating multirecurrent pterygia with symblepharon and motility restriction.

BACKGROUND: Treatment of eyes with multirecurrent pterygia associated with severe symblepharon and motility restriction is challenging. A combined surgical procedure of intraoperative mitomycin C, amniotic membrane transplantation and conjunctival limbal autograft was applied for treating such eyes. METHODS: Seven eyes of seven patients who had previously undergone an average of four operations for pterygial removal and who manifested recurrent pterygia associated with severe symblepharon and motility restriction were involved in this retrospective study. The surgical procedures involved clearing fibrovascular membrane over the cornea, extensive excision of epibulbar fibrovascular tissue to the bare sclera, application of 0.02% mytomycin C onto the bare sclera for 5 min and transplantation of preserved human amniotic membrane and conjunctival limbal autograft. RESULTS: Postoperatively, all seven eyes showed rapid epithelialization on the corneal surface in 3-5 days and, on the conjunctival surface, in 10-18 days. For a mean follow-up period of 22.4+/-6.1 months, six eyes recovered deep fornices, smooth and stable ocular surface and full ocular motility without recurrence. One eye showed regrowth of fibrovascular tissue and motility restriction but less severe than before surgery. No complication was noted due to mitomycin C. CONCLUSIONS: Combined intraoperative mitomycin C, amniotic membrane graft and limbal conjunctival autograft are successful approaches for treating multirecurrent pterygia with severe symblepharon to restore the ocular surface integrity and prevent recurrence.

Fungal spectrum identified by a new slide culture and in vitro drug susceptibility using Etest in fungal keratitis.

PURPOSE: To investigate the spectrum of fungal species causing keratitis and to test antifungal drug susceptibility to each isolate using Etest. METHODS: Microbial cultures were performed for patients who were clinically diagnosed with fungal keratitis between September 2002 and July 2004. Modified slide culture was established to identify the fungal species of the isolates. Etest (AB BIODISK, Solna, Sweden) was applied to determine the antifungal agent susceptibility of each isolate to itraconazole, fluconazole, and amphotericin B in vitro, respectively. RESULTS: Among 73 eyes of 73 patients with clinical diagnosis of fungal keratitis, 61 strains of fungi were isolated from 61 eyes. The rate of positive culture was 81.3% of all cases. The spectrum of fungal species involved: 58 (95.1%) isolates of filamentous fungi, including the two most common genera-Fusarium (n = 33, 54.1%) and Aspergillus (n = 9, 14.8%),-followed by 16 (26.2%) isolates of other genera of filamentous fungi such as Alternaria (n = 3, 4.9%), Trichophyton (n = 3, 4.9%), Curvularia (n = 2, 3.3%), Chrysosporium (n = 2, 3.3%), Acremonium (n = 2, 3.3%), and Scedosporium (n = 1, 1.6%), 1 (1.6%) yeast of Candida, as well as two (3.3%) dimorphic fungi of Blastomyces and Sporothrix isolate each. Three filamentous fungi of the isolates failed to be identified according to the information provided by slide culture. The results of Etest showed that 20 (60.6%) isolates of Fusarium were susceptible to amphotericin B, whereas all of them were resistant to itraconazole and fluconazole. All nine (100%) isolates of Aspergillus were sensitive to itraconazole, whereas four (44.4%) of them were sensitive to amphotericin B, and only two (22.2%) of them were sensitive to fluconazole. Seventeen (89.5%), 13 (68.4%), and 10 (52.6%) isolates of the remaining 19 organisms were sensitive to amphotericin B, itraconazole, and fluconazole, respectively. CONCLUSIONS: Fusarium and Aspergillus are the most frequent pathogenic organisms in causing fungal keratitis, whereas other species of fungi can also cause corneal infection. In vitro Etest for assessing antifungal drug susceptibility is a simple and practical method and may provide referential information for clinical consideration of choosing antifungal agents to treat fungal keratitis.

[Optical penetrating keratoplasty in eyes with severe keratomycosis after therapeutic keratoplasty].

OBJECTIVE: To observe and evaluate secondary optical penetrating keratoplasty (PKP) in the eyes with severe keratomycosis after therapeutic keratoplasty using cryopreserved donor tissue. METHOD: There were 35 eyes in 35 cases for whom therapeutic PKP using cryopreserved donor tissue had been carried out to treat severe fungal keratitis. At least 6 months after the PKP, when the fungal infection had been eradicated and ocular inflammation had resolved, a secondary optical penetrating keratoplasty was further performed for visual rehabilitation. If the crystal lens was found to be opaque, it was also simultaneously removed through extracapsular cataract removal with an IOL implantation. Surgical or postoperative complications, visual outcome, graft rejection episode, and graft survival were analyzed. RESULTS: Among 35 eyes, 18 eyes received optical PKP only, and 17 received combined optical PKP with ECCE and IOL implantation. During the follow-up period of 6.8 - 36.8 (15.7 +/- 7.6) months after optical PKP, 3 eyes experienced graft failure due to immunologic rejection and 32 retained clear grafts. Postoperative visual acuity was 0.4 or better in 24 eyes, and 0.1 or better in 32 eyes, and less than 0.1 in 3 eyes. CONCLUSIONS: Optical PKP performed in the eyes after therapeutic PKP due to severe fungal keratitis can recover satisfactory visual acuity, with low frequency of graft rejection, long-term graft survival and few complications.