M.F. de la Paz

Sterility control and long-term eye-bank storage of cultured human limbal epithelial cells for transplantation

Background/aims: To assess sterility of cultured human limbal epithelial cells (HLEC) and to investigate the viability, morphology and phenotype of cultured HLEC following 2 and 3 weeks of organ culture storage. Methods: HLEC cultured on amniotic membranes were stored in organ culture medium in a closed container at 23°C. Sterility of storage media was tested using a Bactec 9240 blood culture instrument (Becton Dickinson, Maryland) for incubation and periodic reading. Viability was analysed by calcein-acetoxymethyl ester/ethidium homodimer-1 assay, morphology by light microscopy and cellular phenotype by immunohistochemistry. Results: No microbial contamination was observed after 1 week’s storage. Viability of cultured HLEC was 87.9 (SD 6.4)% and 52.7 (13.1)% after 2 and 3 weeks of storage, respectively, compared with 98.8 (2.6)% before storage (p<0.001). The multilayered structure was preserved in 70% of cultures following 2 weeks of storage but lost after 3 weeks. A less differentiated phenotype was maintained. Conclusion: This study is the first to verify the sterility of HLEC cultures prior to transplantation. Although a slight decrease in viability was observed following 2 weeks of storage, the HLEC sheets remain acceptable, whereas 3 week’s storage was unsatisfactory.

Pronóstico visual y complicaciones posquirúrgicas en queratoprótesis de Boston tipo 1

OBJECTIVE To describe the visual outcome of patients who underwent Boston type 1 keratoprosthesis (KPro1) implantation, and describe serious sight-threatening post-operative complications. METHODS We performed an analysis of the clinical records of all patients who underwent Boston keratoprosthesis implantation (BKI)in our institution from May 2006 to February 2011. RESULTS A total of 41 eyes of 37 patients were included in the final analysis, of whom 22 (59.45%) were male and 15 were (40.54%) female. The mean age was 56.44 years (range 2-90). The most frequent diagnoses were bullous keratopathy, autoimmune diseases, such as Stevens-Johnson syndrome (SJS)/Lyell syndrome (LS), and aniridic keratopathy. The mean number of previous keratoplasties (PK) was 2.36 (range 0-8), the mean number of previous non-PK surgeries was 1.58 (range 0-9). The mean follow-up time was 22.17 months (range 3-46). The mean best corrected visual acuity (BCVA) logMAR before surgery was 2.05 (range 1.10-2.52), and the mean best corrected visual acuity achieved after surgery was 1.16 (range 0.08- 2.70). The most frequent complication was the formation of retroprosthetic membrane (RPM) which appeared in 22 (53.65%) eyes. Of these, 6 (27.27%) appeared after another surgery. Fourteen (63.63%) RPM required treatment, an average of 1.71 (range 1-4) laser YAG applications were performed, and surgical membranectomy was performed in 3 patients. Eleven (26.82%) eyes showed chorioretinal adhesion problems, 6 (14.63%) of which occurred after follow-up of BKI surgery. Infectious complications occurred in 7 (17.07%) cases; 2 (4.87%) patients had infectious keratitis and 5 (12.19%) endophthalmitis. CONCLUSIONS Visual function improved in most patients. Those with prior multiple ocular surgeries and alterations of systemic immunity such as SJS, LS, and diabetes mellitus are at increased risk for serious sight-threatening complications, such as RPM, chorioretinal detachment and infection. Nevertheless, we consider KPro as an effective alternative in patients with multiple ocular pathology and imminent risk of rejection of a new KP.

Insuficiencia limbar secundaria a múltiples inyecciones intravítreas

CASE REPORT A 68 year-old male with idiopathic choroidal polypoidal vasculopathy received seven intravitreal injections of bevacizumab in the supero-nasal quadrant. He later developed a localized limbal stem cell deficiency which required limbal transplantation from the same eye. Two months after surgery, a slight improvement of visual acuity was noted, along with an intact ocular surface, clear cornea and decreased inflammation. DISCUSSION Repeated surgical trauma on the limbus and surrounding areas may damage the limbal stem cells, giving rise to a corneal epitheliopathy due to iatrogenic limbal deficiency. Repeated intravitreal injections may be considered as one of the possible causes.

Limbal stem cell deficiency following multiple intravitreal injections

Abstract Case report A 68 year-old male with idiopathic choroidal polypoidal vasculopathy received seven intravitreal injections of bevacizumab in the supero-nasal quadrant. He later developed a localized limbal stem cell deficiency which required limbal transplantation from the same eye. Two months after surgery, a slight improvement of visual acuity was noted, along with an intact ocular surface, clear cornea and decreased inflammation. Discussion Repeated surgical trauma on the limbus and surrounding areas may damage the limbal stem cells, giving rise to a corneal epitheliopathy due to iatrogenic limbal deficiency. Repeated intravitreal injections may be considered as one of the possible causes.

Queratitis neurotrófica posciclofotocoagulación transescleral con láser diodo

OBJECTIVE To study the relationship between treatment with diode laser transscleral cyclophotocoagulation and development a neurotrophic keratitis due to the damage of the sensitive corneal innervation. METHODS A study was conducted on 5 eyes of 5 patients who were treated with diode laser transscleral cyclophotocoagulation and soon developed neurotrophic ulcers. Personal characteristics of the patients were collected, as well as refraction and risk factors for corneal hypoesthesia, and the parameters of the laser used in the surgery. RESULTS It was found that the 5 patients had predisposing factors of corneal hypoesthesia prior to surgery (chronic use of topical beta blockers, surgery with corneal incisions, diabetes mellitus, or corneal dystrophies); however none had developed neurotrophic keratitis until the cyclophotocoagulation was performed. It also showed that 4 of them were highly myopic, and they all were treated with high laser parameters (with an average of 2880 mW for 3s at an average surface of 275°), triggering neurotrophic ulcers between 10 and 35 days after surgery. CONCLUSION Neurotrophic keratitis is a rare complication that can occur after diode laser transscleral cyclophotocoagulation, secondary to the damage of the long ciliary nerves. The emergence of this disorder can be triggered by the existence of previous risk factors, including high myopia, thus it is important to respect the recommended treatment parameters to prevent the development of this disorder.

Crecimiento de novo de un hemangioma capilar conjuntival

CLINICAL CASE A 22-year-old woman patient, diagnosed with an inclusion cyst of the conjunctiva in the nasal sector of the left eye, who after 2 shot/needle injections in the lesion came to our clinic with a dense subconjunctival hemorrhage in four quadrants and with severe pain. After excision biopsy, a capillary hemangioma of the conjunctiva was diagnosed. DISCUSSION Conjunctival capillary hemangioma is mainly a benign lesion, asymptomatic and mostly congenital in origin, its progression or de novo growth is rare in adulthood.

Effect of Transportation on Cultured Limbal Epithelial Sheets for Worldwide Treatment of Limbal Stem Cell Deficiency

Limbal stem cell deficiency can be treated with transplantation of cultured human limbal epithelial cells (LEC). It can be advantageous to produce LEC in centralized labs and thereafter ship them to eye clinics. The present study used transport simulations of LEC to determine if vigorous shaking during transport altered the viability, morphology and phenotype during a 4 day-long storage of LEC with a previously described serum-free storage method. Inserts with LEC cultured on amniotic membranes were sutured to caps inside air-tight containers with generous amounts of 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES)-buffered minimal essential medium (MEM). The containers were distributed among the following testing conditions: 6 hours with full containers, 36 hours with full containers, 36 hours with container three quarters full of medium, and 36 hours with container full of medium containing a shear-protecting agent (Pluronic-F68). Compared to stored, but non-transported controls, no statistically significant changes in viability and immunohistochemical staining were observed. The epithelial sheets remained intact. However, an air-liquid interface in the containers reduced the number of desmosomes and hemi-desmosomes compared to the controls. In conclusion, cultured LEC sheets appear to endure vigorous shaking for at least 36 hours if the container is full.

Amiloidosis tarsal unilateral

CASE REPORT A 23-year-old man presented with a unilateral left ptosis of two years onset. After ineffective treatment for vernal conjunctivitis for several months, the biopsy demonstrated tarsal amyloidosis. Surgical resolution of the case was anatomically and functionally satisfactory. DISCUSSION Ocular amyloidosis is a rare disease that must be suspected with infiltrative mass lesions. Biopsy is the diagnostic method. Treatment should be individualised.

Boston Keratoprosthesis (Type I): visual prognosis and complications

Purpose to describe the functional results and sight threatening post‐operative complications of Type I Boston Keratoprosthesis.

Evolution of cataract surgery: Barraquer experience

Purpose To describe the evolution of cataract surgery at the Barraquer Eye Center