Distribution of TGFBI variants in patients with early onset glaucoma.

Purpose: To describe a novel association of TGFBI variants with congenital glaucoma in a family with GAPO (growth retardation, alopecia, pseudoanodontia, and progressive optic atrophy) syndrome, as well as among other unrelated cases of juvenile onset open-angle glaucoma (JOAG) and primary congenital glaucoma (PCG). Methods: This study of one family of GAPO with congenital glaucoma and three unrelated patients with JOAG analyzed a common link to glaucoma pathogenesis. Three girls with GAPO syndrome born to consanguineous parents in a multi-generation consanguineous family were identified. Two of the girls had congenital glaucoma in both eyes, while the elder sibling (a 10-year-old female) had features of GAPO syndrome without glaucoma. Results: A genetic evaluation using whole exome sequencing revealed a novel homozygous ANTXR1 mutation in all three affected siblings with GAPO. No other mutations were detected in the genes associated with glaucoma. A rare missense variant in the TGFBI gene was shared in the two siblings with congenital glaucoma and GAPO syndrome. We found three other unrelated patients with JOAG and one patient with primary congenital glaucoma with no known glaucoma causing gene mutations, but having four different missense variants in the TGFBI gene. One of these patients with JOAG had familial granular corneal dystrophy. Molecular dynamic simulations of TGFBI and 3-D structural models of three of its variants showed significant alterations that could influence TGFBI protein function. Conclusions: The possibility that variations in the TGFBI gene could have a possible role in the pathogenesis of congenital and juvenile onset open-angle glaucomas needs further evaluation.

Treatment Decision-making of Cracked Teeth: Survey of American Endodontists.

AIM: The aim of this study is to survey the opinion of endodontists in the United States regarding their approach to treatment of cracked teeth. MATERIALS AND METHODS: A survey assessing the opinions of 3,500 endodontists in the United States was administered over a 4-week period. It consisted of a hypothetical clinical case and eight different treatment scenarios. Participants were asked to select one of the two treatment options: (A) informing the patient of the presence of a crack, reinforcing the tooth, and continuing with endodontic treatment; and (B) extraction of the tooth followed by replacement with an implant supported crown or fixed partial denture. RESULTS: When the crack extended across the distal marginal ridge with no associated probing depths, 92.65% endodontists preferred to continue with endodontic treatment and reenforcing the tooth. When the crack extended across the distal marginal ridge and was associated with a probing depth of 6 mm, 80% preferred extraction of the tooth. When the crack extended across the mesial marginal ridge up to the orifice of the mesiolingual canal with no associated probing depth, 82.78% preferred to continue with the treatment. When the crack was associated with a 6-mm probing depth, 83.79% preferred tooth extraction. When the crack extended across the mesial marginal ridge and down into the mesiolingual canal with/without associated probing depth, 91.13% and 63.54% preferred tooth extraction, respectively. When the crack involved both mesial and distal marginal ridges and extended across the pulp chamber, 79.74% preferred tooth extraction. When a split tooth was presented, 98.48% preferred tooth extraction. CONCLUSION: It appears that the presence of a 6-mm periodontal pocket is considered an important factor by most American endodontists when deciding whether to preserve the cracked tooth or extract it. CLINICAL SIGNIFICANCE: There is no consensus among dentists regarding the best approach to treat cracked teeth. Conclusive studies evaluating clinical approaches of endodontists regarding treatment of cracked teeth are lacking. Therefore, surveyed opinion of endodontists in the United States regarding their approach to treatment of cracked teeth was done to try to reach the best clinical decision regarding this dilemma.

Mucopolysaccharidosis.

Mucopolysaccharidosis are group of inherited metabolic diseases caused by the absence or malfunctioning of lysosomal enzymes resulting in accumulation of glycosaminoglycans. Over time this accumulation damages cells, tissues, and organs. There are seven types of MPS and 13 subtypes that are associated with multiple organ systems, such as the respiratory, liver, spleen, central nervous systems, arteries, skeletons, eyes, joints, ears, skin, and/or teeth. The various types share some common ocular features that differ in terms of the severity of the affection. Visual loss in MPS patients is varied and can be due to corneal clouding, glaucoma, retinopathy, and optic neuropathy. The primary focus of this review is on changes in the cornea and anterior segment in MPS patients, including clinical and novel investigative modalities, current surgical management, effects of systemic therapy like hematopoietic stem cell transplants (HSCT)and enzyme replacement therapy (ERT), as well as significant research developments.

Evaluation of 0.3% gatifloxacin hydrochloride in decontamination of donor corneas.

PURPOSE: To evaluate demographic, clinical, and microbiological profile of eye donors and efficacy of 0.3% gatifloxacin hydrochloride in microbial decontamination of donor corneas. METHODS: About 513 donors and 1,026 corneas received at National Eye Bank of a tertiary care hospital during 1-year period were analyzed prospectively in this randomized clinical trial. The donor eyes were graded and treated with 5% povidone-iodine, 0.4% amikacin sulphate, and 0.3% gatifloxacin hydrochloride. The parameters evaluated were death enucleation time (DET), grading of donor corneas, microbiological profile of culture organisms, and their sensitivity to various antibiotics. RESULTS: Mean DET was 6.29±5.7 hours. Forty one percent eyes were optical grade corneas and the majority of donors (38.5%) had accidental deaths. Good grade eyes were maximum with DET of <1 hour and were comparable between 0-6 hours and 6-12 hours. About 57.6% (591/1026) eyes were culture positive; most common organisms were Pseudomonas spp (53%) and Coagulase-negative Staphylococci (24%). Culture positivity reduced significantly after treatment with povidone iodine and amikacin (P=0.002, right eye; P=0.004; left eye) and decreased further with use of gatifloxacin (P=0.001). Pseudomonas (93%), Coagulase-negative Staphylococci (96.3%), Staphylococcus aureus (90.5%), enterococci and gram-negative bacilli were sensitive to gatifloxacin. Pseudomonas spp which were multidrug-resistant were sensitive to polymyxin-B. CONCLUSIONS: Gatifloxacin hydrochloride in addition to amikacin sulphate is beneficial for donor eye decontamination. Polymyxin-B may be used for multidrug-resistant Pseudomonas spp.

Noninvasive ocular surface analyzer as an adjunct in diagnosis and estimating prevalence of meibomian gland dysfunction: Hospital-based comparative study.

Purpose: To assess the role of noninvasive ocular surface analyzer (OSA) in workup of meibomian gland dysfunction (MGD) and to estimate hospital-based prevalence of MGD using this objective device. Methods: The study recruited 113 consecutive participants attending the ophthalmology outpatient department of a tertiary care hospital. All participants were administered a symptom questionnaire. Participants underwent a comprehensive ocular examination, including slit-lamp biomicroscopy and meibomian gland expression. Lipid layer thickness (LLT), noninvasive tear breakup time (NIBUT), tear meniscus height (TMH), and meibomian gland loss (MGL) were assessed using OSA. The presence of either or both reduced/absent meibum secretion and cloudy to toothpaste-like secretion was diagnosed as MGD. Results: Prevalence of total MGD was 57.52% (95% confidence interval [95% CI]: 48.3%-66.8%) and that of symptomatic MGD was 42.5% (95% CI: 33.2%-51.7%). Prevalence of total and symptomatic MGD was highest in those aged ≥50 years (P < 0.001 and P = 0.004, respectively). Computer vision syndrome increased the odds of symptomatic MGD (odds ratio [OR]: 4.3). NIBUT and MGL significantly differed in MGD and non-MGD groups (P = 0.023 and P < 0.001, respectively). LLT significantly differed between asymptomatic and symptomatic cases (P = 0.033). MGL >25% increased the odds of having MGD (OR: 19.1). Significant negative correlations were observed between MGL and NIBUT (P = 0.04) and between MGL and LLT (P = 0.02). MGL demonstrated the highest diagnostic accuracy for MGD (AUC = 0.827, sensitivity = 75.4%, specificity = 85.4%, cut-off value: ≥26%). Conclusion: MGD is a common disorder in adults attending the ophthalmology outpatient services of a tertiary eye care hospital. Incorporating noninvasive OSA in clinical practice can aid in rapid and reliable measurements of MGD-related parameters.

Cellular response of limbal epithelial cells on electrospun poly-ε-caprolactone nanofibrous scaffolds for ocular surface bioengineering: a preliminary in vitro study.

PURPOSE: The aim of this study was to develop a synthetic stromal substrate for limbal epithelial cell (LEC) expansion that can serve as a potential alternative substrate to replace human amniotic membrane (HAM). METHODS: Nanofibers were fabricated using 10% poly-ε-caprolactone (PCL) solution dissolved in trifluoroethanol (TFE) via an electrospinning process. Nanofibers were characterized for surface morphology, wetting ability, pore size, mechanical strength, and optical transparency using scanning electron microscopy (SEM), contact angle measurement, microtensile tester, and UV-Vis spectrophotometer, respectively. The human corneal epithelial (HCE-T) cell line was used to evaluate the biocompatibility of nanofibers based on their phenotypic profile, viability, proliferation, and attachment ability. Subsequently, human LECs were cultivated on biocompatible nanofibers for two weeks and their proliferation capability analyzed using MTT ((3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, a yellow tetrazole)) proliferation assay. Immunofluorescent (IF) staining and reverse transcriptase polymerase chain reaction (RT-PCR) were performed to check the molecular marker expression; SEM was used to study the morphology. RESULTS: The average fiber diameter of PCL was 132±42 nm. Pore size varied from 0.2 to 4 microns with a porosity of 85%. The tensile strength of the PCL membrane was 1.74±0.18 MPa (Mega Pascal); strain was 30.08±2.66%. The water contact angle was 90°. Biocompatibility results indicated that the polymer surface was highly biocompatible, as HCE-T cells could favorably attach and proliferate on the polymer surface. SEM figures showed that the corneal epithelium was firmly anchored to the polymer surface via a continuous cell sheet and was able to retain a normal corneal phenotype. MTT assay confirmed that cells were metabolically active on nanofibers (p<0.05) and gradually increased in their number for up to two weeks. IF and RT-PCR results revealed no change in the expression profile of LECs grown on nanofibers when compared to those grown on glass coverslips and human amniotic membrane (HAM). Confocal microscopy illustrated that cells infiltrated the nanofibers and successfully formed a three-dimensional (3D) corneal epithelium, which was viable for two weeks. CONCLUSIONS: Electrospun nanofibers provide not only a milieu supporting LEC expansion, but also serve as a useful alternative carrier for ocular surface tissue engineering and could be used as an alternative substrate to HAM.

Microbiological profile of donor corneas: a retrospective study from an eye bank in north India.

PURPOSE: To study the microbiological profile and the antibiotic sensitivity of the microorganisms cultured from donor corneas and evaluate with respect to the decontamination protocols during the study period. METHODS: Retrospective review of microbiological culture reports of 2,291 donor eyes sent from January 2000 to December 2003. Donor corneas were treated with 0.4% gentamicin (phase I), 1% povidone-iodine and 0.4% gentamicin (phase II), and 5% povidone-iodine with 0.4% amikacin (phase III). Limbal swabs taken by the eye bank technician after antimicrobial decontamination of donor eyes and sent to the microbiology laboratory for culture were tracked. The positive culture reports with antibiotic sensitivity pattern were studied and results correlated with decontamination protocols. RESULTS: Staphylococcus epidermidis, Pseudomonas aeruginosa, and Alkaligenes faecalis were the 3 most common isolates overall. The percentage of positive bacterial cultures reduced when we used 5% povidone-iodine and 0.4% amikacin (31.3%) as compared to 1% povidone-iodine and 0.4% gentamicin (61.8%) (P < 0.001). Addition of povidone-iodine decreased positive fungal cultures from 6% in phase I to 2.2% in phase II (P < 0.01) and 1.9% in phase III (P < 0.01). Amongst the total bacterial cultures sent after antimicrobial treatment, Gram-positive bacteria decreased from 38.6% (64/166) in phase I to 27.6% (402/1457) and 10.8% (72/668) in phases II and III, respectively (P < 0.001), and Gram-negative bacteria increased from 10.2% (17/166) to 18.8% (274/1457) and 19.8% (132/668), respectively (P = 0.02). Amongst the positive bacterial cultures, the proportion of Gram-positive bacteria decreased from 79.0% (64/81) in phase I to 59.5% (402/676) and 35.3% (72/204) in phases II and III, respectively (P < 0.001), and that of Gram-negative bacteria increased from 21.0% (17/81) to 40.5% (274/676) and 64.7% (132/204), respectively (P < 0.001). CONCLUSIONS: Different combinations of antimicrobials failed to eliminate all organisms, especially Gram-negative bacteria from the donor corneal surface. The increase in proportion of Gram-negative bacteria despite the use of amikacin combined with povidone-iodine suggests a need to find a better combination for decontamination of donor corneal tissue.

Piezoelectric harvest of osteo-odonto-lamina in modified osteo-odonto keratoprosthesis: A maxillofacial perspective.

PURPOSE: We describe the piezo-osteotomy feasibility in rehabilitation and harvesting of osteo-odonto lamina in modified osteo-odonto keratoprosthesis (MOOKP) surgery. Surgery was evaluated regarding operative technique and success of the keratoprosthesis (KPros) in terms of perception to light (+ve) to finger counting (+ve). MATERIALS AND METHODS: This retrospective cohort study included 12 patients undergone MOOKP surgery procedures. Harvesting of osteo-odonto-lamina was performed using piezosurgical osteotomy during 2007-2012. RESULTS: The mean follow up was 34 months (range 24-48 months). Of the 12 patients six patients had vision ≥6/12, four patients had vision <6/12, but >6/60 and one patients had vision ≤6/60. KPros was retained and functional in all the eyes after a minimum follow up of 24 months. There was mucosal overgrowth over the optical cylinder occurred in two cases over 4 years follow up which was corrected with trimming. Postoperative complication at donor site was seen in three cases, two cases with exposure of root of adjacent teeth and oro antral fistula in one. CONCLUSIONS: This study suggests that piezoelectric harvest of osteo-odonto-lamina is a valuable surgical option in patients undergoing MOOKP surgery, resulting in high success rate with less complication.

The osteo-odonto-keratoprosthesis (OOKP).

The osteo-odonto-keratoprosthesis (OOKP), although described over 40 years ago, remains the keratoprosthesis of choice for end-stage corneal blindness not amenable to penetrating keratoplasty. It is particularly resilient to a hostile environment such as the dry keratinized eye resulting from severe Stevens-Johnson syndrome, ocular cicatricial pemphigoid, trachoma, and chemical injury. Its rigid optical cylinder gives excellent image resolution and quality. The desirable properties of the theoretical ideal keratoprosthesis is described. The indications, contraindications, and patient assessment (eye, tooth, buccal mucosa, psychology) for OOKP surgery are described. The surgical and anaesthetic techniques are described. Follow-up is life-long in order to detect and treat complications, which include oral, oculoplastic, glaucoma, vitreo-retinal complications and extrusion of the device. Resorption of the osteo-odonto-lamina is responsible for extrusion, and this is more pronounced in tooth allografts. Regular imaging with spiral-CT or electron beam tomography can help detect bone and dentine loss. The optical cylinder design is discussed. Preliminary work towards the development of a synthetic OOKP analogue is described. Finally, we describe how to set up an OOKP national referral center.

Surface-modified electrospun poly(epsilon-caprolactone) scaffold with improved optical transparency and bioactivity for damaged ocular surface reconstruction.

PURPOSE: The purpose of this study was to modify and functionalize the surface of synthetic poly-ε-caprolactone (PCL) nanofibrous scaffolds to improve their biocompatibility in order to provide better "cell-substrate" interaction. METHODS: Poly-ε-caprolactone solution was electrospun and its surface functionality was modified by helium-oxygen (He/O2) plasma discharge. Scaffolds were characterized for their morphology, wetting ability, mechanical strength, and optical properties by using scanning electron microscopy (SEM), water contact angle measurement, tensile strength, and ultraviolet-visible (UV-Vis) spectrophotometer, respectively. The biocompatibility of nanofibers was explored by culturing human corneal epithelial (HCE-T) cell line. Subsequently, human limbal epithelial cells (LECs) were cultured to evaluate the bioactivity. Cell proliferation was checked by MTT (3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Immunofluorescent staining and reverse transcription-polymerase chain reaction were done to check the gene expression; SEM was used to study the morphology. RESULTS: Plasma-treated and untreated scaffolds showed almost similar morphology and tensile strength. Water contact angle measurement and optical transparency data showed that the plasma-treated PCL (pPCL) exhibited significantly improved wettability and transparency as compared to the untreated PCL scaffolds. Biocompatibility results indicated that both scaffolds are biocompatible in terms of cell survival and proliferation. However, pPCL showed better cell adhesion and proliferation. Results supported that LEC cultured on pPCL scaffolds had enhanced cell adhesion and proliferation, in comparison to untreated PCL. Gene expression study showed cultures were able to retain their normal phenotype on both scaffolds. CONCLUSIONS: The hydrophilicity of the surface achieved by plasma treatment effectively enhanced the transparency and promoted the biocompatibility of scaffolds. These nanofibers may act as biological cues for endorsing ocular surface engineering.

Intrascleral fibrin glue intraocular lens fixation combined with Descemet-stripping automated endothelial keratoplasty or penetrating keratoplasty.

PURPOSE: To evaluate the outcomes of intrascleral haptic fixation of an intraocular lens (IOL) with fibrin glue combined with penetrating keratoplasty (PKP) or Descemet-stripping automated endothelial keratoplasty (DSAEK) for aphakic or pseudophakic bullous keratopathy (BKP). SETTING: Rajendra Prasad Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, New Delhi, India. DESIGN: Case series. METHODS: Eyes with BKP had combined PKP or DSAEK with fibrin glue-assisted intrascleral posterior chamber (PC) IOL fixation; PKP was performed in eyes with a corneal scar and DSAEK in eyes without a scar. The parameters evaluated were corrected distance visual acuity (CDVA), anterior segment biomicroscopy, intraocular pressure, central corneal thickness (CCT), and IOL status. Intraoperative events and postoperative complications were recorded. RESULTS: The study evaluated 11 patients (11 eyes). Intrascleral fixation of a PC IOL with PKP or DSAEK was successfully performed in all eyes; PKP was performed in 6 eyes (54.54%) and DSAEK in 5 eyes. The mean CDVA improved from 1.95 logMAR ± 0.29 (SD) to 0.40 ± 0.16 logMAR (P<.001). The mean CCT was 0.741 ± 0.71 mm preoperatively and 0.579 ± 0.20 mm postoperatively (P<.001). There were no cases of intraoperative or postoperative IOL decentration or other complications. CONCLUSIONS: Fibrin glue-assisted intrascleral fixation of a PC IOL combined with DSAEK or PKP was a safe, effective method to manage BKP with aphakia or malpositioned IOLs. The IOL fixation was strong enough to sustain the manipulation required for corneal procedures. FINANCIAL DISCLOSURE: No author has a financial or proprietary interest in any material or method mentioned.

Sutureless patch graft for sterile corneal melts.

An innovative technique to seal large paracentral corneal melts is being presented. Two patients with sterile corneal melts sized 3-5 mm in diameter were treated with a full-thickness patch graft punched using a dermatological trephine and secured with the fibrin-aprotinin biological tissue adhesive, supplanting the need for sutures. The defect was adequately sealed, and successful tectonic support with postoperative best-corrected visual acuity of 6/24 was achieved in both cases, circumventing the need for an emergency full-thickness penetrating keratoplasty.

Pyruvate inhibits galactosemic changes in cultured cat lens epithelial cells.

An attempt was made to maintain cat lens epithelial cells (CLEC) in culture and study the morphology, growth and survival of these cells in vitro. The influence of incorporation of galactose (30 mM) into the culture medium on the morphology and biochemistry of CLEC in the primary culture was then investigated. To establish the effect of galactose on CLEC, various biochemical parameters associated with galactosemic cataract such as aldose reductase (AR), Na+K+ATPase, glutathione, polyol and soluble/insoluble proteins were estimated after 24 h of incubation. The effect of pyruvate (5 mM), a 'physiological antioxidant', on the changes induced by galactose in CLEC was studied. CLEC in culture showed regular hexagonal cells with prominent nuclei. The CLEC culture attained confluency in 11 days during primary culture and semiconfluency in 14 days in two subsequent passages. Vacuolization and significantly raised AR activity, polyol levels and insoluble protein contents were observed; they had no effect on Na+K+ATPase and soluble protein after 24 h of incubation in the culture medium with galactose. Supplementation of pyruvate (5 mM) resulted in a lesser number of vacuoles together with a positive modulation of these parameters.