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To compare the changes encountered in corneal biomechanics and aberration profile following accelerated corneal collagen cross-linking (CXL) using hypo-osmolar and iso-osmolar riboflavin in corneal thicknesses of <400 and >400 microns, respectively. Methods: This is a prospective, interventional, comparative study involving 100 eyes of 75 patients with progressive keratoconus. Eyes were divided into two groups based on corneal thickness: group 1 included eyes with a corneal thickness of <400 microns who underwent hypo-osmolar CXL, and group 2 included eyes with a corneal thickness of >400 microns who underwent iso-osmolar CXL. Corneal biomechanical and aberration profiles were evaluated and compared between groups. Results: In group 1, all higher-order aberrations (HOA) except secondary astigmatism significantly decreased from baseline; however, in group 2, only coma and trefoil decreased. The corneal resistance factor and corneal hysteresis significantly improved in both groups, which was significantly greater in group 2 than in group 1. The change in inverse radius, deformation amplitude, and tomographic biomechanical index was significantly improved in group 2 as compared to group 1. Conclusion: Improvement in corrected distance visual acuity and decrease in HOA were significantly better in the hypo-osmolar CXL group; however, the improvement in biomechanical strength of the cornea was significantly better in the iso-osmolar group. [ABSTRACT FROM AUTHOR]

The effect of riboflavin-induced collagen photocrosslinking on the scaffold printing accuracy, namely on the area of formed niches and filament thickness, as well as on the degradation time and biocompatibility of scaffolds, has been studied. The thickness of the filaments in the riboflavin-modified scaffolds was 13–29% less and the niche area was 23–40% larger than in the control group without riboflavin. The riboflavin addition reduced weight loss and increased the scaffold degradation time in the buffer solution and collagenase solution by 1.2–1.3, and 1.4–2.0 times, respectively, depending on the time of exposure to UV light. The existence of an optimal illumination interval was established. Cultivation of mesenchymal stem cells on a riboflavin-crosslinked scaffold for a week did not lead to a decrease in their viability and proliferation rate. [ABSTRACT FROM AUTHOR]

Periodontitis, a chronic inflammatory disease caused by microbial communities carrying pathogens, leads to the loss of tooth-supporting tissues and is a significant contributor to tooth loss. This study aims to develop a novel injectable cell-laden hydrogel consisted of collagen (COL), riboflavin, and a dental light-emitting diode (LED) photo-cross-linking process for periodontal regeneration. Utilizing α-SMA and ALP immunofluorescence markers, we confirmed the differentiation of human periodontal ligament fibroblasts (HPLFs) into myofibroblasts and preosteoblasts within collagen scaffolds in vitro. Twenty-four rats with three-wall artificial periodontal defects were divided into four groups, Blank, COL_LED, COL_HPLF, and COL_HPLF_LED, and histomorphometrically assessed after 6 weeks. Notably, the COL_HPLF_LED group showed less relative epithelial downgrowth (p < 0.01 for Blank, p < 0.05 for COL_LED and COL_HPLF), and the relative residual bone defect was significantly reduced in the COL_HPLF_LED group compared to the Blank and the COL_LED group (p < 0.05). The results indicated that LED photo-cross-linking collagen scaffolds possess sufficient strength to withstand the forces of surgical process and biting, providing support for HPLF cells embedded within them. The secretion of cells is suggested to promote the repair of adjacent tissues, including well-oriented periodontal ligament and alveolar bone regeneration. The approach developed in this study demonstrates clinical feasibility and holds promise for achieving both functional and structural regeneration of periodontal defects. [ABSTRACT FROM AUTHOR]

This study was conducted to evaluate scleral remolding-related gene expression after scleral collagen cross-linking (SCXL) using ultraviolet A (UVA) and riboflavin in lens-induced myopia (LIM) guinea pigs. A total of 100 4-week-old pigmented guinea pigs were randomly divided into five groups (n = 20): SCXL + LIM, LIM, SCXL, Sham, and Control. Refraction, anterior chamber depth (ACD), lens thickness (LT), vitreous chamber depth (VCD), and axial length (AL) were measured using streak retinoscope and A-scan ultrasonography. SCXL was performed using 0.1% riboflavin solution and 365 nm UVA irradiation. Lens-induced myopia was achieved by wearing −10 D concave lenses. Quantitative real-time PCR (qPCR) and western blot were used to measure mRNA and protein levels, respectively. Myopia was successfully induced in the LIM group, while myopic refraction was higher and ACD and AL were shorter in SCXL + LIM compared with LIM, suppressing myopia progression. The scleral COL1A1 mRNA levels were significantly decreased and MMP2 and ACTA2 mRNA levels were significantly increased in LIM compared with other groups, while COL1A1 mRNA levels were increased and MMP2 and ACTA2 mRNA levels were decreased in SCXL + LIM compared with LIM. The scleral COL1A1 protein levels were significantly increased at 1 week and 4 weeks and MMP2 protein levels were significantly decreased at 1 week in SCXL compared with SCXL + LIM, LIM and Control. MMP2 protein levels were significantly decreased in SCXL + LIM and SCXL compared with LIM at 4 weeks. The differences in TGFB1, BMP2, CCN2, ITGA2, and ITGB1 mRNA levels and ACTA2 protein levels between the five groups were not significantly different. SCXL using UVA and riboflavin could influence the expression of scleral remolding-related genes, including COL1A1, MMP2, TIMP2, and ACTA2, and thus contribute to improving collagen synthesis and reducing collagen degradation and might have an effect on slowing myopia progression. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate published data regarding riboflavin (RF) as a cross-linker for improved adhesive bond strength to dentin and to analyze previous studies for optimal concentration of riboflavin range suitable for dentin bond. Saliva and distilled water were used as storage media and aging time was 24 h and 6 months. Results of meta-analysis were synthesized using a statistical method of inverse variance in random effects with a 95% Confidence Interval (CI). Cochrane review manager 5.4.1 was used to determine results of the meta-analysis. In total, 3172 articles were found from search databases "PubMed", "Scopus", and "Google Scholar". Six of the fifteen studies were eligible for meta-analysis. Micro tensile strength shows significant improvement with the addition of riboflavin (p < 0.05) compared to without the addition of riboflavin from with 95% CI. A significant difference has been found in micro tensile bond strength between use of the riboflavin cross-linker and without use of the riboflavin crosslinker in the dentin adhesive system. With a 95% confidence interval (CI), the I2 for micro tensile strength was 89% with strong heterogeneity, Chi2 = 44.76, df = 5 (p < 0.00001), and overall effect size is Z = 2.22 (p = 0.03) after immediate aging. Chiang et al. 2013 shows maximum mean differences which is 38.50 [17.93–59.07]. After 6 months of aging in distilled water or artificial saliva micro tensile bond strength has been increased with the addition of riboflavin (p < 0.05). It can be clearly seen that pooled effect and 95% CI did not cross the line of no effect. With a 95% confidence interval (CI), the I2 for micro tensile strength was 96% with strong heterogeneity, Chi2 = 117.56, df = 5 (p < 0.00001), and overall effect size is Z = 2.30 (p = 0.02). Subgroup analysis proved a similar effect of distilled water and artificial saliva as storage media on micro tensile bond strength after incorporating riboflavin as a collagen crosslinker. An artificial saliva aged forest plot also showed considerable heterogeneity with I2 = 96%; Tau2 = 257.32; Chi2 = 94.37; df = 2 (p < 0.00001); test for overall effect, Z = 1.06 (p = 0.29). Riboflavin prior to or with bonding is recommended to improve the bonding of different adhesive systems. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effects of scleral collagen cross-linking (SXL) using riboflavin and ultraviolet A (UVA) light on the scleral thickness of different regions and expression of matrix metalloproteinase 2 (MMP-2) and membrane-type MMP-1 (MT1-MMP) in guinea pigs with lens-induced myopia. Methods: Forty-eight 4-week-old guinea pigs were assigned to three groups (n = 16 per group): SXL group, lens-induced myopia (LIM) group, and control group. The sclera of the right eye of the guinea pig in the SXL group was surgically exposed, riboflavin was dropped on the treatment area for 10 minutes before the 30-minute UVA irradiation. The same surgical procedure was performed in the LIM group without UVA irradiation. The -10.00 D lenses were then placed on the right eyes of guinea pigs in the SXL and LIM groups for six weeks. The control group received no treatment. The left eyes were untreated in all groups. The ocular axial length (AXL) and refraction were measured at 4 weeks and 10 weeks of age. 10-week-old guinea pigs were sacrificed, and the right eyes were enucleated and evenly divided for preparation of hematoxylin and eosin (HE) stained sections, quantitative real-time polymerase chain reaction (qPCR) and western blotting. The scleral thickness of different regions was measured on HE stained sections. The temporal half of the sclera was harvested to measure the expression of MMP-2 and MT1-MMP by qPCR and western blotting. Results: The AXL was significantly shorter, and the degree of myopic refraction was significantly lower in the SXL group than those in the LIM group at 10 weeks of age. The scleral thickness of the cross-linked area was significantly greater in the SXL group than that of the corresponding area in the LIM group, while the scleral thickness of the untreated nasal side was not significantly different between the SXL group and the LIM group. The expression of MMP-2 and MT1-MMP of the cross-linked sclera was significantly downregulated compared with that of the corresponding area in the LIM group. Conclusion: Riboflavin/UVA SXL could slow myopia progression and thicken the cross-linked sclera in guinea pigs, which might be related to the downregulation of MMP-2 and MT1-MMP expression during the scleral remodeling process. [ABSTRACT FROM AUTHOR]

The purpose of this article is to evaluate the effectiveness of collagen crosslinking of filtering blebs with 0.1% riboflavin stimulated with ultraviolet A radiation in a case of a thin-walled filtering bleb with leakage following an episode of late blebitis. The time to cessation of the bleb leakage was studied, as well as the intraocular pressure, the visual acuity, and the presence of adverse effects during 4 years of follow-up after treatment. A single session of crosslinking was effective in resolving the leakage of the filtering bleb. There were no adverse effects. The intraocular pressure and the visual acuity remained stable after 4 years of follow-up. Collagen crosslinking with 0.1% riboflavin stimulated with ultraviolet A radiation may prove to be a simple, painless, non-invasive, and repeatable treatment of the leaking bleb with the aim of strengthening the collagen fibers of the conjunctiva and avoiding more invasive surgical treatments. [ABSTRACT FROM AUTHOR]

To evaluate long-term outcomes of corneal collagen crosslinking (CXL) using riboflavin and UV-A irradiation and to determine when to repeat CXL. In this retrospective consecutive interventional case series 131 eyes of 131 patients (95 male, 36 female, mean age 29.7 ± 11.4 years) between 2006 and 2016 received standard CXL (Dresden protocol, epithelium-off) for progressive keratoconus. Corrected distance visual acuity (CDVA) and corneal tomography (K1, K2, Kmax) were repeatedly recorded 1 year (n = 103 eyes) to 10 years (n = 44) postoperatively. Only one eye per patient was included. Paired t-test or Wilcoxon matched-pairs signed rank test was used for parametric and nonparametric data, respectively. 1–3 years preoperatively, median K2 significantly increased by 1.1 D (p < 0.001). Postoperatively, median K2 increased by 0.1 D after 1 year, then decreased over the remaining postoperative period by 0.85 D (p = 0.021). Kmax fluctuated without significant change. Median apical corneal thickness decreased by 16 µm (p = 0.012) after 5 years and then returned to preoperative values. Mean CDVA showed a significant improvement (decrease in logMAR 0.08 after 10 years, p = 0.010). CXL non-responders, defined by a postoperative increase in Kmax>2 D, increased from 16% after 5 to 33% after 10 years. Risk factors for non-response were young age, high astigmatism (>4.3 D), thin cornea (<480 µm), poor initial visual acuity (CDVA ≥0.3 D), and atopic dermatitis. 4 eyes were re-treated 3–4 years after first CXL without complications and keratoconus stabilized thereafter. CXL can slow or stop keratoconus progression. However, as the number of responders declines after 5 years, especially patients with risk factors may require re-treatment. [ABSTRACT FROM AUTHOR]

Purpose: To study the demographics and clinical profile of keratoconus (KC) presenting in pre-teen children in India.Methods: This was a retrospective case series conducted as a single-institutional study at a tertiary eye center in India. A total of 586 eyes from 294 KC patients (aged 12 years or less) without any active comorbid conditions of the eye were included in the study. Slit-lamp biomicroscopy was used to document the clinical signs of KC. Information on age; gender; reason for consultation; family history; history of allergy, atopy, and eye rubbing; manifest refraction; uncorrected and best-corrected distance visual acuity (UCVA and BCVA, respectively); clinical presentation; and contact lens usage were also analyzed, along with data on types of medical and surgical treatments for KC and their outcomes.Results: The mean age of this pediatric KC patient cohort was 9.3 ± 1.8 years, and there was a male (70%) preponderance. Baseline mean UCVA, BCVA, steep keratometry, and flat keratometry were 0.86 ± 0.58 logMAR, 0.44 ± 0.38 logMAR, 54.82 ± 8.4 D, and 48.21 ± 9.5 D, respectively. Progression, necessitating collagen crosslinking (CXL), was noted in 12.7% eyes. Post-CXL, visual and topographic parameters remained stable without any complications till 6 months posttreatment. However, in eyes that did not undergo CXL, significant progression over time (P < 0.001) was observed. A keratoplasty was required in 2.3% eyes.Conclusion: KC was present at an advanced stage in 25% of the pre-teens in our series, and therefore, it is an important diagnostic entity when a refractive error is diagnosed, even in very young children. [ABSTRACT FROM AUTHOR]

Purpose: To analyze the 5-year results of accelerated corneal collagen crosslinking (CXL) for progressive keratoconus and identify preoperative characteristics predictive of visual and topographic outcomes.Methods: A prospective interventional case series. Nineteen eyes of 19 patients receiving accelerated CXL with settings of 18 mW/cm2 for 5 min were included. Clinical and topographic parameters were assessed. Linear regression and logistic regression were used to compare the R2 and odds ratio (OR), respectively, between baseline characteristics and postoperative outcomes.Results: Corrected distance visual acuity (CDVA) remained stable from 0.28 ± 0.21 to 0.25 ± 0.18 logMAR (P = 0.486). The mean cylindrical refraction was stable (P = 0.119). The maximal keratometry (Kmax) decreased from 61.99 ± 10.37 to 59.25 ± 7.75 D (P < 0.001), flattening in the flattest and steepest meridians and mean keratometry were also observed (P ≤ 0.040). The mean anterior elevation at the apex reduced from 21.42 ± 16.69 to 18.53 ± 12.74 μm (P = 0.013) and changes in posterior elevation were non-significant (P = 0.629). Preoperative Kmax best predicted the postoperative change in Kmax (R2 = 0.55, P < 0.001) compared to the other baseline characteristics (P ≤ 0.028), whereas preoperative CDVA was the only significant predictor of postoperative change in CDVA (R2 = 0.41, P = 0.003). Accelerated CXL is less likely to fail in eyes with a steeper preoperative Kmax (OR = 0.74, P = 0.040) or greater posterior elevation at the apex (OR = 0.91, P = 0.042).Conclusion: Kmax significantly decreased following accelerated CXL. Eyes with worse preoperative CDVA and higher Kmax were more likely to have an improvement in visual acuity and corneal flattening. [ABSTRACT FROM AUTHOR]

Corneal collagen crosslinking (CXL) is a commonly used minimally invasive surgical technique to prevent the progression of corneal ectasias, such as keratoconus. Unfortunately, riboflavin/UV-A light-based CXL procedures have not been successfully applied to all patients, and result in frequent complications, such as corneal haze and endothelial damage. We propose a new method for corneal crosslinking by using a Ruthenium (Ru) based water-soluble photoinitiator and visible light (430 nm). Tris(bipyridine)ruthenium(II) ([R u (bpy) 3 ]2+) and sodium persulfate (SPS) mixture covalently crosslinks free tyrosine, histidine, and lysine groups under visible light (400–450 nm), which prevents UV-A light-induced cytotoxicity in an efficient and time saving collagen crosslinking procedure. In this study, we investigated the effects of the Ru/visible blue light procedure on the viability and toxicity of human corneal epithelium, limbal, and stromal cells. Then bovine corneas crosslinked with ruthenium mixture and visible light were characterized, and their biomechanical properties were compared with the customized riboflavin/UV-A crosslinking approach in the clinics. Crosslinked corneas with a ruthenium-based CXL approach showed significantly higher young's modulus compared to riboflavin/UV-A light-based method applied to corneas. In addition, crosslinked corneas with both methods were characterized to evaluate the hydrodynamic behavior, optical transparency, and enzymatic resistance. In all biomechanical, biochemical, and optical tests used here, corneas that were crosslinked with ruthenium-based approach demonstrated better results than that of corneas crosslinked with riboflavin/ UV-A. This study is promising to be translated into a non-surgical therapy for all ectatic corneal pathologies as a result of mild conditions introduced here with visible light exposure and a nontoxic ruthenium-based photoinitiator to the cornea. Keratoconus, one of the most frequent corneal diseases, could be treated with riboflavin and ultraviolet light-based photo-crosslinking application to the cornea of the patients. Unfortunately, this method has irreversible side effects and cannot be applied to all keratoconus patients. In this study, we exploited the photoactivation behavior of an organoruthenium compound to achieve corneal crosslinking. Ruthenium-based organic complex under visible light demonstrated significantly better biocompatibility and superior biomechanical results than riboflavin and ultraviolet light application. This study promises to translate into a new fast, efficient non-surgical therapy option for all ectatic corneal pathologies. [Display omitted] [ABSTRACT FROM AUTHOR]

In connective tissues there is a clear link between increasing age and degeneration. Advanced glycation end-products (AGEs) are believed to play a central role. AGEs are sugar-induced non-enzymatic crosslinks which accumulate in collagen with age and diabetes, altering tissue mechanics and cellular function. Despite ample correlative evidence linking collagen glycation to tissue degeneration, little is known how AGEs impact cell-matrix interactions, limiting therapeutic options. One reason for this limited understanding is that AGEs are typically induced using high concentrations of ribose which decrease cell viability, making it impossible to investigate cell-matrix interactions. The objective of this study was to develop a system to trigger AGE accumulation while maintaining cell viability. Using cell-seeded high density collagen gels, we investigated the effect of two systems for AGE induction, ribose at low concentrations (30, 100, and 200 mM) over 15 days of culture and riboflavin (0.25 and 0.75 mM) induced with blue light for 40 seconds (riboflavin-465 nm). We found ribose and riboflavin-465 nm treatment produces fluorescent AGE quantities which match and/or exceed human fluorescent AGE levels for various tissues, ages, and diseases, without affecting cell viability or metabolism. Interestingly, a 40 second treatment of riboflavin-465 nm produced similar levels of fluorescent AGEs as 3 days of 100 mM ribose treatment. Riboflavin-465 nm is a promising method to trigger AGEs on demand in vivo or in vitro without impacting cell viability and offers potential for unraveling the mechanism of AGEs in age and diabetes related tissue damage. [ABSTRACT FROM AUTHOR]

Scleral photocrosslinking is increasingly investigated for treatment of myopia and glaucoma. In this study a computational model was developed to predict crosslinking efficiency of visible/near infrared photosensitizers in the sclera. Photocrosslinking was validated against riboflavin corneal crosslinking experimental studies and subsequently modeled for the sensitizer, methylene blue, administered by retrobulbar injection to the posterior sclera and irradiated with a transpupillary light beam. Optimal ranges were determined for treatment parameters including light intensity, methylene blue concentration, injection volume, and inspired oxygen concentration. Additionally, sensitivity of crosslinking to various parameters was quantified. The most sensitive parameters were oxygen concentration in the injection solution, scleral thickness, and injection reservoir thickness (i.e., injection volume). [Display omitted] • Scleral stiffening by photocrosslinking was modeled for glaucoma/myopia treatments. • The model was validated against published riboflavin corneal photocrosslinking data. • Scleral crosslinks and treatment parameters were modeled over space and time. • Optimal ranges for treatment parameters for photocrosslinking were predicted. • Sensitivity analysis suggests tissue parameters were most impactful to crosslinking. [ABSTRACT FROM AUTHOR]

Background: This study aimed to analyze if the outcome after corneal crosslinking (CXL) in progressive keratoconus patients depends on the stage at which the procedure is performed. This knowledge would help to improve success of CXL and to define surgery indications in those patients.Methods: In this retrospective study, 124 consecutive eyes of 100 patients with progressive keratoconus undergoing corneal CXL at the University Eye Hospital Tübingen were included. The eyes were graded according to modified Krumeich stages depending on induced myopia or astigmatism, corneal radii, minimum corneal thickness, and morphological changes. The observation period covered November 2008 to September 2018. Preoperatively, 12 and 24 months after CXL, the best corrected visual acuity (BCVA) was determined and astigmatism as well as tomographic parameters (Kmax, Kmin, central corneal thickness (CCT), minimum corneal thickness (MCT)) were measured by means of a Scheimpflug camera system.Results: BCVA results showed significant differences between the modified Krumeich stages at 12 months (p = 0.014) and at 24 months postoperatively (p = 0.032). Also, astigmatism differed significantly among the stages at 24 months after CXL (p = 0.023). However, no significant differences regarding astigmatism were detectable after 12 months. In terms of Kmax, Kmin, CCT, and MCT, no significant differences between the Krumeich stages were observed.Conclusions: BCVA showed a significantly higher improvement after CXL in the early stage of keratoconus compared to a higher stage. However, the postinterventional tomographic values did not differ significantly between the different modified Krumeich stages. The significantly higher improvement in BCVA after CXL in the early stage might indicate that earlier intervention provides a higher subjective benefit to the individual. Further studies with larger sample sizes are needed to confirm these findings. [ABSTRACT FROM AUTHOR]

Purpose: To evaluate the long-term visual, refractive, tomographic, and aberrometric outcomes of corneal collagen crosslinking (CXL) with or without hypoosmolar riboflavin solution in the treatment of progressive keratoconus patients with thin corneas. Methods: Charts of consecutive progressive keratoconus patients with thinnest corneal thickness less than 470 µm who underwent corneal collagen CXL with or without hypoosmolar riboflavin solution and using a standardized protocol for treatment and examinations were analyzed retrospectively. The indication for hypoosmolar riboflavin use was a central corneal thickness less than 400 µm as measured by ultrasound pachymetry after epithelial debridement and before exposure to ultraviolet A (UVA) light. Uncorrected distance visual acuity (UDVA), best spectacle-corrected distance visual acuity (CDVA), manifest refraction, slit lamp biomicroscopy, corneal tomography, corneal aberrometry, and endothelial cell counts were evaluated at baseline and yearly at all postoperative follow-up examinations until month 36. The outcomes of corneal CXL procedure performed using hypoosmolar riboflavin were compared to those performed using the standard procedure. Results: Twenty-three eyes (19 patients) were treated using hypoosmolar riboflavin application, and 30 eyes (28 patients) were treated using the standard procedure. Compared to baseline, the mean UDVA, CDVA, and keratometric readings improved statistically significantly in both groups at postoperative year 3, without any statistically significant between-group differences. Progression was not observed in any patient eye in either group. No significant endothelial cell loss and no sight threating complication were observed in any patient eye. Conclusion: At 3 years follow-up, the safety and efficacy of CXL using hypoosmolar riboflavin solution seems to be similar to that of standard CXL in progressive keratoconic eyes with thin corneas. The visual, refractive, keratometric, tomographic and aberrometric outcomes of the two procedures were comparable, as well. [ABSTRACT FROM AUTHOR]

Background: This study aimed to evaluate the clinical outcomes and assess preoperative characteristics that may predict outcomes in keratoconus 1 year after accelerated transepithelial corneal cross-linking (ATE-CXL).Methods: This prospective study included 93 eyes of 84 consecutive keratoconus patients with 1-year follow-up after ATE-CXL. Preoperative characteristics included corneal astigmatism, anterior chamber depth, anterior chamber volume, radius of curvature, posterior elevation, central corneal thickness (CCT), thinnest corneal thickness, steepest meridian keratometry, flattest meridian keratometry, and the maximum keratometry (Kmax). Data were obtained preoperatively and at 1, 3, 6, and 12 months postoperatively. The patient eyes were grouped into 3 subgroups according to CCT and Kmax values to observe the changes of keratoconus progression.Results: All patients were successfully operated without complications at any follow-up time point. Mean changes of Kmax from baseline at 6 and 12 months were - 0.60 ± 2.21 D (P = 0.011) and - 0.36 ± 1.58 D (P = 0.030), respectively. Eyes with a thinner CCT and higher Kmax values exhibited a tendency for topographic flattening of ≥1.0 D (P = 0.003; P = 0.003). In the subgroup comparison, the Kmax values decreased significantly at 6 and 12 months after ATE-CXL in the group with CCT ≤ 450 μm (P = 0.018 and P = 0.045); the Kmax values of the group with Kmax > 65.0 D decreased significantly at 6 months postoperatively (P = 0.025).Conclusion: ATE-CXL is a safe and effective treatment for keratoconus patients. Patients with thinner CCT and higher Kmax values are more likely to benefit from ATE-CXL. [ABSTRACT FROM AUTHOR]

Purpose: To monitor the changes in the ABCD grading system during a one-year follow-up after a corneal cross-linking (CXL) procedure.Methods: This prospective study included 30 eyes of 25 patients with keratoconus, who received the CXL treatment. The patients with a history of ocular trauma or surgery and other corneal pathology were excluded from the study. The patients were examined at the baseline visit and followed up at 3, 6, and 12 months after the CXL. All the patients underwent a standard CXL procedure with visual acuity and Scheimpflug tomography testing at each visit. The corneal parameters and ABCD grading were monitored throughout the follow-up period.Results: There were no significant changes of parameter A and anterior radius curvature (ARC) in the ABCD grading system. Parameters B and D showed progression postoperatively, with an improvement of parameter D on the final visit. Parameter C showed a statistically significant increase at all three post-CXL visits, but a constant gradual decrease in the value over time.Conclusion: The ABCD grading system can be very useful in monitoring the progression of keratoconus (KC), but it can also help in monitoring the efficacy of corneal cross-linking. The anterior surface parameters in the ABCD grading system did not show progression in the post-CXL period, and parameters C and D showed improvement and stability a year after the procedure. [ABSTRACT FROM AUTHOR]

Purpose: To find the intraocular lens (IOL) power calculation changes before and after isotonic collagen cross-linking (CXL) in keratoconus patients.Methods: : Thirty-five eyes of 25 patients who underwent isotonic CXL were included. The cases included conventional CXL (n = 16), accelerated CXL (n = 7), contact lens-assisted CXL (CACXL) (n = 9), accelerated CACXL (n = 3). All underwent ocular biometry (IOL master), corneal topography (Orbscan II), and simulated keratometry (Orbscan II) preoperatively and 1-year post CXL. Change in best-corrected visual acuity (BCVA), axial length (AL), simulated keratometry (Sim K), anterior chamber depth (ACD), and IOL power were analyzed in the overall data and then grouped based on flattening (Group A) and no flattening (Group B) of Sim K value post CXL procedure.Results: For the overall data, there was no significant change in IOL power (P = 0.05) at the end of 1 year, BCVA showed a significant increase (P < 0.01), and Sim K reading showed a statistically significant flattening (P = 0.001); ACD and AL showed insignificant change. In intergroup comparison, there was no statistically significant change in IOL power. However, in Group A, a significant change in BCVA and Sim K values was observed. In both groups (Group A and Group B), IOL power was found to be negatively correlated with AL and Sim K values.Conclusion: Isotonic CXL did not affect IOL power calculation at the end of 1 year. However, significant change in BCVA and sim K reading was noted. [ABSTRACT FROM AUTHOR]

Purpose: To evaluate the efficacy of CXL in treating fungal keratitis as an adjuvant therapy.Methods: Detailed clinical examination microbiological investigation was performed. Twenty fungal keratitis patients were recruited and randomized into two groups: group 1 (n= 11, standard antifungal), group 2 (n=9, corneal collagen crosslinking with standard antifungal). Corneal scraping and tear samples collected were subjected to real-time PCR targeting ITS, TLR analysis and cytokine analysis.Results: The mean time for complete resolution of ulcer for group 2 was significantly shorter compared to group 1 and the final mean BCVA was better for group 2. Expression of IL-1β, IL-8, IFN-γ significantly decreased immediately post CXL in group 2 patients. Significant downregulation of TLR 6, TLR-3, TLR-4 was observed 3-days post CXL compared to group 1 patients.Conclusion: Adjuvant effect of CXL was significant in treating fungal keratitis compared to standalone antifungal treatment. [ABSTRACT FROM AUTHOR]

Background: Keratoconus is a condition characterized by biomechanical instability of cornea presenting in a progressive asymmetry and bilateral way. Corneal collagen crosslinking with riboflavin and UVA is a technique that has influenced the treatment of Keratoconus by arresting the progression. Aims and Objectives: The aim of the study was to assess the short-term efficacy and safety of collagen cross-linking in Keratoconus. Materials and Methods: This prospective, nonrandomized, interventional case study was conducted at Aravind Eye hospital and PGI, Puducherry from June 2012 to April 2013. Statistical analysis was done using repeated measures ANOVA and Bonferonn's using IBM SPSS Inc. 20.Version. Results: A total of 30 patients were enrolled in this study and standard cross linking procedure was done. It was found that UCVA, BSCVA both improved about 40%. Spherical Equivalent reduced by 0.7 D, cylinder reduced by 0.8 D and total astigmatism decreased by about 0.4 D. Steepest K value reduced by 2 D, Irregularity and astigmatism at 3 and 5 mm also changed significantly (P<0.05). Conclusion: Collagen cross-linking arrests the progression of Keratoconus. It seems to improve the UCVA and BCVA and it is a safe and effective procedure. [ABSTRACT FROM AUTHOR]

Introduction: Keratoconus (KC) is the most common corneal ectatic disorder with a prevalence of about 1 in 2,000 people. KC is a progressive, non-inflammatory, almost bilateral and asymmetric of corneal. Corneal collagen cross-linking (CXL) with riboflavin and ultraviolet- A (UVA) is a new technique of mechanical stability of the cornea and prevention of keratoconus progression. Purpose: Determining the relationship between changes in corneal density before and after collagen cross-linking surgery in patients with keratoconus Materials and Methods This prospective study was performed on 45 eyes of 23 patients with keratoconus undergoing corneal collagen cross-linking treatment at Imam Khomeini Hospital in Ahvaz in 2019. Corneal densitometry (CD) and pachymetry were measured using Pentacam before CXL and at 3, 6 and 12 months after CXL. Also, all of the patients were evaluated in terms of best corrected visual acuity (BCVA), during follow-up. Results: In this study, 23 people including 12 men (54.2%) and 11 women (45.8%) with a mean age of 25.2 ± 4.64 years in the age range of 18-34 years participated. The mean value of corneal density in total layer over the annular diameters 0-2 mm was 19.10±5.58, 22.31±5.64, 24.28±6.37 and 20.89±5.73 for before, 3 months, 6 months and 12 months after CXL, respectively (P = 0.001). The mean value of corneal density in ant layer over the annular diameters 0-2 mm was 22.38±8.38, 27.46±9.63, 28.07±11.28 and 24.53±9.21for before, 3 months, 6 months and 12 months after CXL, respectively (P = 0.001). The mean value of corneal pachymetry in thinnest location was 426.2±34.7 µm, 412.5±42.6 µm, 419.2±37.3 µm and 428.3±36.9 µm for before, 3 months, 6 months and 12 months after CXL, respectively (P = 0.392). The pre-CXL value of BCVA was significantly lower than the post-CXL values in 12 months (0.10 ± 0.19, 0.15 ± 0.10 and 0.019 ± 0.15, respectively, P<0.0001). In general, Densitometry values increases and corneal thinnest location decreses during the 3-6 months after CXL and then return to baseline preoperative values in 12 months. There was a significant improvement in best corrected visual acuity (BCVA) before and 12 months after surgery (P<0.0001). Conclusions: CXL can increase corneal densitometry and decrease corneal thinnest location during the postoperative 6 months period. Densitometric values and corneal thickness return to baseline preoperative values to a large extent during 6-12 months postoperatively. The patient's BCVA increased over 12 months but no correlation was observed between this increase and densitometric changes. [ABSTRACT FROM AUTHOR]

Background: Collagen cross-links contribute to the mechanical resilience of the intervertebral disc (IVD). UVA-light-activated riboflavin-induced collagen crosslinking (UVA-CXL) is a well-established and effective ophthalmological intervention that increases the mechanical rigidity of the collagen-rich corneal matrix in Keratoconus. This study explores the feasibility, safety and efficacy of translating this intervention in reinforcing the IVD. Methods: Annulus fibrosus (AF) cells were isolated from bovine IVDs and treated with different combinations of riboflavin (RF) concentrations (0.05–8 mM) and UVA light intensities (0.3–4 mW/cm2). Metabolic activity (resazurin assay), cell viability (TUNEL assay), and gene expression of apoptosis regulators C-FOS and PT5 were assessed immediately and 24 hours after treatment. Biomechanical effects of UVA-CXL on IVDs were measured by indentation analysis of changes in the instantaneous modulus and by peel-force delamination strength analysis of the AF prior and after treatment. Results: Different intensities of UVA did not impair the metabolic activity of AF cells. However, RF affected metabolic activity (p < 0.001). PT53 expression was similar in all RF conditions tested while C-FOS expression decreased 24 hours after treatment. Twenty-four hours after treatment, no apoptotic cells were observed in any condition tested. Biomechanical characterizations showed a significant increase in the annular peel strength of the UVA-CXL group, when compared to controls of UVA and RF alone (p < 0.05). UVA-CXL treated IVDs showed up to 152% higher (p < 0.001) instantaneous modulus values compared to the untreated control. Conclusion: This is the first study on UVA-CXL treatment of IVD. It induced significantly increased delamination strength and instantaneous modulus indentation values in intact IVD samples in a structure–function relationship. RF concentrations and UVA intensities utilized in ophthalmological clinical protocols were well tolerated by the AF cells. Our findings suggest that UVA-CXL may be a promising tool to reinforce the IVD matrix. [ABSTRACT FROM AUTHOR]

Purpose: The aim of this study was to test a method for estimating corneal rigidity before and after cross-linking (CXL) using a Schiøtz tonometer.Methods: The study was performed in the Kyiv City Clinical Ophthalmological Hospital "Eye Microsurgical Center", Ukraine. This was a prospective, consecutive, randomized, masked, case-by-case, clinical study. Corneal rigidity, indicated by the gradient (G) between lg applied weight and corresponding lg scale reading during Schiøtz tonometry, were obtained by increasing (A-mode) then reducing (D-mode) weights by two operators [A] in keratoconus, post-CXL and control subjects for estimation of (i) interoperator and (ii) intersessional errors, (iii) intergroup differences; [B] before and after CXL. Central corneal thickness CCT was measured by scanning slit pachymetry. ANOVA, t tests, linear regression were the statistical tools used.Results: Average interoperator difference (ΔG) was -0.120 (SD = ±0.294, 95%CI = -0.175 to -0.066). A significant correlation between ΔG and the mean of each pair of G values was found (r = -0.196, n = 112, P = 0.038). Intersessional differences in mean G values were insignificant (P > 0.05). There was a significant correlation between G at first session (X1) and difference between sessions (ΔG) [Operator 1, ΔG = 0.598x1-0.461, r = 0.601, n = 27, P = 0.009]. Significant intergroup differences in G were found (Operator 1, one-way ANOVA, F = 4.489, P = 0.014). The difference (Δ) between the pre-(X2) and post-CXL treatment G values was significantly associated with the pre-CXL treatment value (Operator 1, Δ = 1.970x2-1.622, r = 0.642, n = 18, P = <.001). G values were correlated with CCT in keratoconus and post-CXL.Conclusion: Corneal rigidity (G) estimated using the Schiøtz tonometer can be useful for detecting changes after CXL. However, G values are linked to CCT, can vary from time-to-time and the procedure is operator dependent. [ABSTRACT FROM AUTHOR]

Purpose: To compare the safety and efficacy of 0.1% riboflavin in two different solutions which is used in corneal collagen crosslinking (CXL) for the treatment of keratoconus. Methods: This was a prospective, randomized, comparison study which included 100 eyes of 61 patients with progressive keratoconus who underwent CXL with riboflavin 0.1% solution as a photosensitizer, using the standard Dresden protocol of using 3mW/cm2 UV-A irradiation for 30 minutes which corresponds to a total energy of 5.4 J/cm2. The recruited patients were divided into 2 groups ie, Flavin Group and Peschke-D group by computer generated randomization. Postoperative examinations were conducted on 1 day, 1 month, 3 months, 6 months and 12 months after the crosslinking. Results: For both groups, the mean manifest spherical equivalent (SE), astigmatism, best corrected distance visual acuity, keratometry values, thinnest pachymetry values, demarcation line depth and endothelial cell density preoperatively and at postoperative 12 months were comparable with no statistically significant differences. At 12 months postoperatively, 62% of eyes in the Flavin group, and 68% of eyes in the Peschke-D group had postoperative manifest SE of within ± 1.00 D. During UV-A exposure, the cornea in the Flavin group showed intraoperative thinning of 112 microns (27%) as compared with a thinning of 108.12 microns (26.5%) observed in the Peschke group (p=1.67) from the initial pachymetry readings. No eye in either group had any immediate or long-term postop vision threatening complications such as infectious keratitis, corneal melt, non-resolving corneal oedema or endothelial decompensation. Conclusion: Both riboflavin solutions were equally safe and effective in the management of progressive keratoconus, and resulted in similar changes in terms of mean manifest spherical equivalent (SE), astigmatism, best corrected distance visual acuity, keratometry values, thinnest pachymetry values, demarcation line depth and endothelial cell density at the end of 12 months postoperatively. Trial Registration Number: Ctri/2019/11/021841 (Www.ctri.nic.in). [ABSTRACT FROM AUTHOR]

Purpose: To analyze the effect of riboflavin UV-A illumination on mRNA and protein expression of healthy (HCFs) and keratoconus human corneal fibroblasts (KC-HCFs), concerning the inflammatory markers NF-κB, iNOS, IL-6, and collagen 1 and 5 (Col 1/Col 5). Methods: Keratocytes were isolated from healthy (n = 3) and keratoconus (KC) corneas (n = 3) and were cultivated in basal medium with 5% fetal calf serum, which resulted in their transformation into human corneal fibroblasts (HCFs/KC-HCFs). Cells underwent 0.1% riboflavin UV-A illumination for 250 s (CXL). NF-κB, iNOS, IL-6, Col 1, and Col 5 expression was investigated by qPCR and Western blot analysis. IL-6 concentration of the cell culture supernatant and cell lysate was determined by ELISA. Results: In untreated KC-HCFs, NF-κB (p = 0.0002), iNOS (p = 0.0019), Col 1 (p = 0.0286), and Col 5 (p = 0.0054) mRNA expression was higher and IL-6 expression was lower (p = 0.0057), than in healthy controls. In HCFs, CXL led to an increased NF-κB (p = 0.0286) and IL-6 (p = 0.0057) mRNA expression. The IL-6 concentration in the cell culture supernatant was increased in HCFs (p = 0.0485) and KC-HCFs (p = 0.0485) after CXL. CXL increased intracellular IL-6 concentration only in KC-HCFs (p = 0.0357). In the HCF group (p = 0.0286), an increased Col 1 mRNA expression after CXL could be observed. Conclusion: Our study confirmed altered gene expression in untreated KC-HCFs compared to untreated HCFs. Riboflavin UV-A illumination affected gene expression only in HCFs. Increased IL-6 concentration in the cell culture supernatant and cell lysate indicate a secondary inflammatory response of HCFs and KC-HCFs to riboflavin UV-A illumination. [ABSTRACT FROM AUTHOR]

Purpose: To review the scientific literature on the comparison of the efficacy of different corneal collagen crosslinking (CXL) protocols for the treatment of progressive keratoconus. Methods: Systematic review of randomized clinical trials (RCTs) on CXL outcomes. A search was carried out using the Cochrane Library, PubMed, EMBASE, Web of Science, Ovid MEDLINE, and Scopus databases. Internal validity was analyzed by applying the filter CASPe (Critical Appraisal Skills Program Spain). Results: The search yielded 1151 articles, and among these, 14 articles met the inclusion and exclusion criteria defined. Conventional (S) crosslinking (CXL) provided better topographic outcomes than transepithelial (TE) CXL, and S-CXL had a better therapeutic effect of corneal flattening than accelerated (A) CXL. The corneal thinning after CXL was lower with hypotonic riboflavin than with riboflavin-dextran. While one study demonstrated a better therapeutic effect of corneal flattening with S-CXL than with A-CXL, another study showed similar results between both techniques. No correlation was found between the depth of the demarcation line and topographic changes, which was not a direct measure of treatment effectiveness. Quality analysis of the literature reviewed yielded a mean score of 8.64, indicating that the RCTs evaluated had an overall acceptable quality. Conclusions: Good-quality RCTs comparing CXL techniques have been conducted, and most of them suggest that epi-off CXL can be considered the standard treatment for progressive keratoconus. TE-CXL and iontophoresis-assisted CXL are mainly indicated in patients with a risk of corneal scarring and patients with pain intolerance, respectively. [ABSTRACT FROM AUTHOR]

Corneal cross-linking (CXL) is a common surgical procedure used to modify corneal biomechanics and stabilize keratoconus progression which is still under discussion. Its side effects, which are mostly related to anatomical unpredictability and stromal exposure, are the reason for the search for new CXL agents. In this work we have quantitatively evaluated the porcine corneal stroma architecture treated with collagen crosslinking agents such as riboflavin solutions and açai extract, using second harmonic generation microscopy. Aimed at evaluating the morphological changes in the corneal stroma after collagen crosslinking under a CXL chemical agent, a tubeness filter based Hessian matrix to obtain a 3D fiber characterization of the SHG images was applied. The results showed a curling effect and shortening of the collagen fibers treated with açai as compared to the control. They also showed a higher degree of clustering of the collagen fibers with larger empty spaces when compared to the other two groups. We believe that studies such as these presented in this paper are a good direct nondestructive and free labeling evaluation technique that allows the observation of morphologic features of corneas treated with new CXL agents. [ABSTRACT FROM AUTHOR]

Contact lens-assisted corneal cross-linking (CACXL) was introduced by Jacob et al. in 2012 for treating thin keratoconic corneas using riboflavin soaked soft contact lens to artificially increase the functional corneal thickness. It is advantageous over other thin corneal cross-linking techniques as it works independent of swelling properties of the cornea, is an epi-off technique and does not require additional time, additional expensive equipments or special solutions. The only additional requirement as compared to all other techniques is a UV barrier-free soft contact lens (SoflensTM, B&L) which is easily available and inexpensive. Advantages include simplicity, easy adaptability, early visual rehabilitation, good visual outcomes, safety, and efficacy. Progression rates are acceptable and the need for re-treatment has been low. CACXL can help regularize corneal shape and may be used in isolation or synergistically with Intracorneal ring segments (ICRS) or Corneal allogenic intrastromal ring segments (CAIRS). It gives about 70% stiffening as compared to standard Dresden protocol CXL in less ideal porcine eye studies. Murine eye models that closely mimic thin corneas and show greater cross-linking effect as compared to porcine eyes may be a better model for evaluation of CACXL, however further studies are needed. Care should be taken in selecting the right kind of contact lens. Proper technique should be followed, especially by confirming thinnest functional pachymetry to be above 400 microns intra-operatively before application of UV-A. The sub-contact lens riboflavin film should be avoided as also an excessively thick supra-contact lens riboflavin film and too many re-applications. [ABSTRACT FROM AUTHOR]

The past two decades have witnessed an unprecedented evolution in the management of keratoconus that demands a holistic approach comprising of inhibiting the ectatic progression as well as visual rehabilitation. The advent of corneal cross-linking (CXL) in the late 1990s resulted in long-term stabilization of the ectatic cornea along with limited reduction in corneal steepening and regularization of corneal curvature. However, CXL as a standalone procedure does not suffice in rehabilitating the functional vision especially in patients who are unwilling or intolerant towards contact lenses. The concept of "CXL plus" was proposed which incorporates adjunctive use of refractive procedures with CXL in order to overcome the optical inefficiency due to corneal irregularity, decrease the irregular astigmatism, correct the residual refractive error and improve functional visual outcome in keratoconus. Several refractive procedures such as conductive keratoplasty (CK), photorefractive keratectomy (PRK), transepithelial phototherapeutic keratectomy (t-PTK), intrastromal corneal ring segments (ICRS) implantation, phakic intraocular lens (PIOL) implantation and multiple other techniques have been combined with CXL to optimize and enhance the CXL outcome. This review aimed to summarize the different protocols of CXL plus, provide guidelines for selection of the optimum CXL plus technique and aid in decision-making for the comprehensive management of cases with primary keratoconus in addition to discussing the future and scope for innovations in the existing treatment protocols. [ABSTRACT FROM AUTHOR]

Background: Keratoconus is a frequent, familiar, bilateral, non inflammatory, and gradually progressive corneal ectasia, which leads to corneal protrusion, irregular astigmatism, and decrease vision. Currently, the corneal cross-linking (CXL) treatment could attain vision improvement. Aim: To investigate the topographic, refractive, and clinical-out-comes after six months CXL in gradually progressing keratoconic eyes. Patients & Methods: A follow-up, randomized, controlled-trial study was used. In this study, 30eyes with progressive-keratoconus (PK) were recruited in the study. Cross-linking was done and achieved by a solution of 0.1% riboflavin mixture with 20% dextran for 15minutes before and during 30minutes time-period of ultraviolet A (UV-A) irradiation (3Mw\cm2). The obtained data were analysed using IBM SPSS software. Results: Statistically significant improvements were noticed in Sim k (at 6months), Kmin (at 6months), spherical power (at 6momths), cylinder power (at 6momths), spherical equivalent [SE] (at 6months), uncorrected visual acuity (UCVA) (at 6months) and best corrected visual acuity [BCVA] (at 6months) in trial group (15patients) when compared with the controls. Conclusions: At 6months, hopeful, promising improvements were reported in Kmax, UCVA, and BCVA. So, long term studies are requested to survey the efficiency of CXL as well as the safety of the repeated procedures. [ABSTRACT FROM AUTHOR]

Purpose: To compare the corneal densitometry changes after transepithelial pulsed (t-ACXL), epithelium-off continuous (c-ACXL) and epithelium-off pulsed (p-ACXL), accelerated corneal crosslinking for patients with progressive keratoconus.Methods: Prospective, interventional case series at a university hospital tertiary referral center. Forty eyes received t-ACXL, 40 eyes received c-ACXL, and 40 eyes received p-ACXL. Corneal tomography, densitometry and visual acuity were evaluated preoperatively and at 1, 3, 6, 12 and 24 months postoperatively.Results: Total densitometry values of the total layer over the annular diameters 0-12 mm expressed in grayscale units (GSU) showed no significant change after t-ACXL and p-ACXL but increased significantly after c-ACXL at 1 month (16.76 ± 2.38, P value < 0.01) and 3 months (16.12 ± 2.22, P value 0.04), returning to baseline levels at month 6 (14.97 ± 2.13, P value 0.99) and remained stable until final follow-up. At 1 month, the mean change in total densitometry value of the t-ACXL group (-0.53 ± 2.08) was significantly lower than the p-ACXL (0.70 ± 1.96 P value 0.04) and c-ACXL (1.97 ± 2.65 P value < 0.01). There was no statistical difference in the mean change in total densitometry between the three groups from 6 to 24 months. In the c-ACXL group, a higher degree of CXL-induced corneal haze at 1 month was observed in patients with steeper preoperative keratometry readings and in individuals where a greater flattening effect was achieved at 24 months.Conclusions: Continuous epithelium-off accelerated cornea crosslinking appears to induce more corneal haze than pulsed epithelium-off and transepithelial pulsed cornea crosslinking in the early postoperative period. Differences resolve by six-months. [ABSTRACT FROM AUTHOR]

Purpose: Determine optimal iontophoresis times for riboflavin delivery to the corneal stroma across different species and compare these to corneal injection. Methods: Ex vivo horse, dog, rabbit, and pig globes were treated with riboflavin administered with either iontophoresis for 2.5‐20 minutes with or without corneal epithelium; or with purpose‐designed precise corneal injection (PCI) application with intact epithelium. Immediately following riboflavin administration, samples were harvested, frozen, and sectioned. Riboflavin penetration was imaged using fluorescence microscopy. Results: Horse samples processed with iontophoresis without epithelium for 2.5, 5, and 7.5 minutes, and processed with intact epithelium for 20 minutes, had mean percent stromal penetration (%SPmean) of 63.4%, 93.8%, 100.0%, and 0.0% (respectively). Dog samples processed with iontophoresis without epithelium for 2.5 and 5 minutes, had %SPmean of 60.7% and 82.1% (respectively). Pig samples processed with iontophoresis for 5 minutes without and with epithelium had %SPmean of 63.3% and 35.1% (respectively). Rabbit samples processed with iontophoresis without epithelium for 2.5 and 5 minutes, had %SPmean of 81.8% and 100.0% (respectively). For all injected volumes, riboflavin was observed spanning throughout the corneal stroma, and lamellar separation was noted surrounding all sites of injection. Conclusions: Both iontophoresis and injection via PCI needles provide efficient and effective means of riboflavin administration in ex vivo horse, dog, rabbit, and pig corneas. Epithelial debridement is required for stromal delivery of riboflavin using iontophoresis in horses. Following epithelial removal, riboflavin penetrated through the horse corneal stroma faster than all other species tested. [ABSTRACT FROM AUTHOR]

Aims: This study aimed to report refractive, topographic, and tomographic outcomes of 2-years corneal collagen cross-linking in patients with progressive keratoconus. Materials and Method: This prospective study was conducted on76 eyes of 40 patients with progressive keratoconus and a corneal thickness of at least 400μm. After the baseline examination, cross-linking was performed using riboflavin 0.1% and ultraviolet irradiation (370 nm, 3 mW/cm2) for 30 min. Uncorrected visual acuity (UCVA), best spectacle-corrected visual acuity (BSCVA), manifest refraction spherical equivalent (MRSE), cylinder, keratometric readings, and corneal thickness were evaluated at baseline, 6, 12, and 24 months after corneal collagen cross-linking. Results: Results showed that at a 24-month follow-up, mean BSCVA, and UCVA had significant improvements. Changes in MRSE and cylinder were not significant during the two years of follow-up. At a 2-years follow-up, the mean values of minimum keratometry, maximum keratometry, and average keratometry decreased significantly to0.75, 0.97, and 0.52 D, respectively, compared to their baseline values. Anterior best fit sphere (BFS) also had significant difference at 24-month follow-up compared to base line evaluation. The central corneal thickness decreased significantly up to38 μ after 2 years. Conclusion: Corneal collagen cross-linking is an effective treatment to stop the progression of keratoconusas shown by reduced keratometry and improved vision. [ABSTRACT FROM AUTHOR]

Purpose: To investigate the corneal epithelial thickness profiles in patients with a confirmed diagnosis of stable and progressive keratoconus.Setting: Studio Italiano di Oftalmologia, Rome, Italy.Design: Observational study.Methods: 86 patients with either stable (n = 52) or progressive (n = 34) keratoconus and 182 healthy controls were enrolled in the study. Disease progression was confirmed by repeated corneal topographies over 1 year follow-up before inclusion in the study. All subjects had full corneal and epithelial thickness mapping taken by spectral domain optical coherence tomography (SD-OCT). The full corneal mapping was investigated by evaluating the central corneal thickness, the thinnest point, the superonasal-inferotemporal thickness difference and the minimum-median thickness difference. The epithelial mapping was investigated by assessing the 2 mm central thickness, the inferior paracentral (2-5 mm) thickness, and the minimum-maximum thickness difference.Results: No significant differences in full corneal mapping were found between stable and progressive keratoconic eyes. Of note, the inferior paracentral region of the corneal epithelium was significantly thinner in progressive (50 ± 3 μm) than stable (53 ± 4 μm) keratoconus (P < 0.001).Conclusions: The SD-OCT corneal epithelial mapping was valuable for detecting local thickness changes in eyes with keratoconus. Monitoring the corneal epithelial changes across the inferior area in patients with keratoconus could be worthy for assessing disease progression. [ABSTRACT FROM AUTHOR]

Background: Corneal collagen cross-linking (CXL) has been used as a treatment option in patients with either keratoconus or iatrogenic corneal ectasia after LASIK. The purpose of corneal cross-linking is to stabilize the corneal stroma and to delay the progression of these two pathologic entities. Objective: The aim of this study is to compare the corneal hysteresis before and after corneal collagen cross-linking using riboflavin and ultraviolet-A light by the epithelial-off technique for treatment of keratoconus. Patients and methods: This study is a single-centre prospective interventional study. It was conducted between April 2018 to December 2018 and included 30 eyes of 18 patients with a mean age 27.08 years (range from 17- 30). They were 12 female eyes and 18 male eyes; all underwent corneal collagen cross-linking using riboflavin and ultraviolet-A light at Kobry El Kobba Military Specialized Eye Hospital. Results: All patients were evaluated preoperatively for their uncorrected corrected visual acuity (UCVA), best corrected visual acuity (BCVA), k-reading, spherical equivalent and pachymetry (using Pentacam), and corneal biomechanical properties (using ocular response analyzer (ORA). The mean age was 27.08±7.86 years, and postoperative follow up was at 1 and 4 months. The mean CH was 7.93±1.79 mmHg preoperatively, 8.27±1.22 mmHg at 1 month, 8.99±1.24 mmHg at 4 months, which show a statistically significant increase in the mean CH pre- and postoperatively, while CRF the other parameter, show a highly statistically significant decrease pre- and postoperatively. Conclusion: CXL is a promising new treatment modality for keratoconus patients and may affect corneal biomechanics. [ABSTRACT FROM AUTHOR]

Importance: Management of cystic bleb leak is difficult. It would be essential to look for a method to strengthen the original pathological conjunctiva and reverse bleb leak. Background: To evaluate the long‐term efficacy and safety of collagen crosslinking in patients with leaking cystic bleb. Design Prospective interventional case series at a university‐based hospital. Participants: Twelve eyes in 12 subjects with late‐onset bleb leak from cystic bleb, without indications for prompt surgical interventions were included. Methods: The subjects underwent crosslinking with 0.1% riboflavin application to bleb surface, followed by ultraviolet irradiation for 30 minutes. The subjects were followed up at baseline and at 1 week, 1 month, 3 months, 6 months post‐treatment and then every 6 months afterwards. Main Outcome Measures: Interval from treatment to cessation of bleb leak, recurrence rate of bleb leak and side effects of treatment. Results: The mean follow‐up after crosslinking was 29.33 ± 12.45 months. Bleb leak subsided in 11 (92%) of 12 patients after a single session of crosslinking, after 1 to 8 weeks (median 3 weeks). Time to leak cessation was significantly correlated with the number of prior glaucoma interventions (R = .71, P = .014). Bleb wall at 3 months was significantly thicker than at baseline (0.70 ± 0.67 vs 0.81 ± 0.62 mm, P = .008). None of the patients experienced any complications. Conclusions and Relevance: Crosslinking achieves resolution of cystic bleb leak which lasts for at least 12 months, without the need of subsequent surgical interventions. Crosslinking is a simple, non‐invasive treatment for bleb leak. It aims to restore the integrity of conjunctiva. [ABSTRACT FROM AUTHOR]

Purpose: To compare the effect of corneal collagen cross‐linking (CXL) on progressive keratoconus using 0.1% riboflavin with either dextran or methylcellulose as the main supplement. Methods: In a comparative case series, CXL was performed in 40 patients (40 eyes) using a riboflavin solution containing either dextran (dextran–riboflavin; n = 20) or methylcellulose (methylcellulose–riboflavin; n = 20). Changes in central corneal thickness (CCT), Scheimpflug tomography, maximal keratometry reading (Kmax), visual acuity (VA) and endothelial cell density (ECD) were recorded. Stromal changes one month after surgery were analysed using optical coherence tomography (OCT) and in vivo confocal microscopy (IVCM). Results: The CCT was significantly higher in the methylcellulose–riboflavin group during the CXL procedure. The IVCM demarcation line depth was 274 ± 80 (SD) μm in the dextran–riboflavin group and 442 ± 80 μm in the methylcellulose–riboflavin group (p < 0.001). Complete absence of keratocytes in the pre‐endothelial stroma was found in none of the corneas treated with dextran–riboflavin and in 42% of the corneas treated with methylcellulose–riboflavin. Visibility of the OCT demarcation line was significantly lower in the methylcellulose–riboflavin group. Kmax and corrected distance visual acuity were improved in the methylcellulose–riboflavin group and stable in the dextran–riboflavin group after 2 years. Endothelial cell density (ECD) was stable in both groups. Conclusion: We found deeper structural changes in the methylcellulose–riboflavin group than in the dextran–riboflavin group. This may be explained by different riboflavin solution properties and raises safety concerns. The study also indicates improved effect using methylcellulose–riboflavin than dextran–riboflavin, possibly explained by deeper stromal CXL effect. [ABSTRACT FROM AUTHOR]

INTRODUCTION: Riboflavin- and ultraviolet (UV)-A-mediated collagen cross-linking of the cornea is a frequently used therapeutic measure for the treatment of progressive keratoconus (PK). First, riboflavin increases cross-linking and second, it serves as a protective shield to other deep ocular structures. However, pharmacogenomic variation in riboflavin efficacy is reported. As the Northeast Indian population represents a genetically diverse group of population compared to mainstream India, we have assessed the efficacy of the procedure in a northeastern population with PK. METHODS: In this study, 78 eyes with PK were included (n = 39 in the treatment arm and n = 39 in the control arm). The primary objective was to evaluate the effect of corneal collagen cross-linking using riboflavin (C3R) (epithelium off) on maximum keratometry. The secondary objectives were evaluation of change in corneal topography parameters, i.e., minimum keratometry (Kmin), simulated keratometry (Sim K), subjective refraction (cylinder power, spherical power, and spherical equivalent [SE]), uncorrected visual acuity (UCVA), best-corrected visual acuity (BCVA), and contrast sensitivity (CS) and safety (intraocular pressure, endothelial cell density, and percentage hexagonality) at 1, 3, and 6 months following C3R procedure. RESULTS: Statistically significant improvement was noted in Kmin(6 months), Sim K (3 and 6 months), cylinder power (3 and 6 months), spherical power (3 and 6 months), SE (3 and 6 months), BCVA (6 months), and UCVA (1, 6 months) in the C3R group (n = 39) when compared to the control group (n = 39). The mean CS decreased at 1 month and gradually improved to achieve statistically significant value at 6 months in the C3R group (P < 0.05). CONCLUSION: Riboflavin-assisted C3R treatment showed promising efficacy in the treatment of PK patients in our population. As the collagen turnover rate of cornea is 2–3 years and the progression of PK is highly variable, we need long-term studies to evaluate the efficacy of C3R over time, requirement of repeat therapy, and safety of repeat cross-linking. [ABSTRACT FROM AUTHOR]

Introduction: The demarcation line (DL) observed since the pioneering crosslink (CLX) protocol at the posterior edge of the cross-linked stroma has been universally accepted as a therapeutic milestone of treatment. Numerous laboratory and clinical CXL studies demonstrate that a deeper DL is associated with a higher amount and saturation level of crosslinks, a more pronounced stiffening effect, and a more durable ectasia stability. Areas covered: A critical revision of laboratory, clinical, and analytical studies on the DL depth supports the significance of the DL as an evaluator of the performance of CLX procedures in terms of biomechanical efficacy and safety avoiding extensive experiments. A mechanical approach based on experimental data shows that the DL depth obtained with different CXL protocols relates with an asymptotic non-linear increasing function to the modified biomechanical corneal stiffness (elastic modulus). Expert opinion: The strong connection between the depth of the DL and the increase of the biomechanical efficacy can be explained by means of UV cross-linking chemical investigations demonstrating that only a limited amount of free reactive collagen residues is involved in the short-wave UV-mediated CXL. Thus, the CXL density can rise only up to an upper boundary value, i.e. the saturation value. [ABSTRACT FROM AUTHOR]

Purpose: To compare the subclinical inflammatory response (as measured by anterior chamber flare) induced after standard (3 mW/cm2, 30 min) and accelerated (18 mW/cm2, 5 min) corneal cross-linking (CXL). Methods: In this comparative, non-randomized study, patients with progressive keratoconus who underwent standard or accelerated CXL were studied. Laser flare photometery (FM-600; Kowa, Tokyo, Japan) was used to measure anterior chamber flare preoperatively and at 1 week, 1 month, 3 months, and 6 months after the procedure. Results: Sixty eyes of 60 patients were studied; 30 eyes in each group. Mean baseline flare values were 4.15 ± 1.19 and 4.57 ± 2.17 ph/ms in standard and accelerated groups, respectively (p = 0.228).and after surgery increased in all follow-up measurements in the both groups similarly (P > 0.05). Conclusion: Both standard and accelerated CXL results in induction of a subclinical inflammatory response that persists up to 6 month. The response was similar between the two groups. [ABSTRACT FROM AUTHOR]

Background. To evaluate the efficacy of phototherapeutic keratectomy (PTK) combined with photorefractive keratectomy (PRK) and riboflavin with ultraviolet-A collagen cross-linking (CXL), performed sequentially on the same day, in the management of corneal ectasia after LASIK. Methods. This retrospective review comprised consecutive patients with corneal ectasia after LASIK. The patients were administered PTK and PRK on the previous corneal flap, and CXL was given on the same day by the same surgeon. The main outcome measures included age, sex, uncorrected distance visual acuity (UDVA), corrected distance visual acuity (CDVA), spherical equivalent refraction, cylinder equivalent refraction, steep and flat keratometries (K), central corneal thickness (CCT), endothelial cell count, corneal haze, and ectasia stability. Mean follow-up period was 6, 12, and 24 months. Results. Sixteen eyes of twelve patients were included in the study. Twenty-four months after administration of PTK combined with PRK and CXL, a significant improvement in UDVA was observed. Mean cylinder equivalent refraction was significantly reduced at 6, 12, and 24 months postoperatively. However, no significant reduction was observed in spherical equivalent refraction. A significant reduction in the flat K and steep K values was observed. No significant change in mean CCT value was observed. Mean endothelial cell count and morphology were unchanged between preoperative and postoperative patients. In addition, no obvious corneal haze was observed. Conclusions. PTK combined with PRK and CXL on the same day is a safe and effective treatment in improving visual acuity in selected patients with corneal ectasia after LASIK. [ABSTRACT FROM AUTHOR]

Abstract Objective Effect of d -alpha-tocopheryl poly(ethyleneglycol)-1000-succinate (VE-TPGS) with riboflavin-5′-phosphate solution on crosslinking of dentine collagen was investigated to analyze collagen's structural integrity. Methods VE-TPGS was added to RF-solution, at RF/VE-TPGS (w/w) ratios of 0.125/0.250 and 0.125/0.500. Demineralized dentine beams were used (10 wt.% phosphoric acid), rinsed using deionized-water and analysed using ELISA (Human MMP2 ELISA; Human CTSK/Cathepsin-K for MMP2 and Cathepsin K analysis). AFM of dentine collagen-fibrils structure was done before and after dentine specimens' placement in mineralization solution and tested after 14 days in artificial saliva/collagenase (AS/Co) solution. The specimens were tested after 24 h in mineralization solution for surface/bulk elastic modulus. Nano-indentation was carried out for each specimen on intertubular-dentine with lateral spacing of 400 nm. Reduced elastic-modulus and nano-hardness were calculated and collagen content was determined using hydroxyproline-assay. Micro-Raman were performed. TEM was carried out to study structural variations of dentine-collagen in artificial-saliva (collagenase). Data were presented as mean ± standard deviation and analyzed by SPSS v.15, by analysis of variance. Results Synergetic effect of VE-TPGS was observed with RF through higher structural integrity of dentine collagen-fibrils shown by TEM/AFM. Superior surface/bulk mechanical stability was shown by nano-indentation/mechanical testing. Improvement in collagenase degradation resistance for hydroxyproline release was observed and lower endogenous-protease release of MMP-2/Cathepsin-K. Raman-analysis analysed chemical interactions between RF and collagen confirming structural-integrity of collagen fibrils after crosslinking. After 24 h mineralization, AFM showed mineral depositions in close association with dentine-collagen fibrils with RF/VE-TPGS formulations. Significance Potential synergetic effect of RF/VE-TPGS was observed by reflection of higher structural integrity and conformational-stability of dentine-collagen fibrils. [ABSTRACT FROM AUTHOR]

Purpose: Contact lens‐assisted corneal cross‐linking (CACXL) has been proposed for the cross‐linking treatment in thin corneas. The aim of this study was to assess the biomechanical efficacy of this treatment. Methods: Post‐mortem porcine eyes were treated with standard cross‐linking and with cross‐linking placing a contact lens soaked with isoosmolar riboflavin solution on the debrided cornea with or without an adherent precorneal riboflavin film of up to 100 μm thickness. Three soft contact lenses (Air Optix Aqua, SofLens and Galifa) with different degrees of hydrophilic properties were tested. After cross‐linking with a surface UVA irradiance of 3 mW/cm² for 30 min (fluence 5.4 J/cm²), a 400 μm deep anterior corneal flap was created using a lamellar rotating microkeratome. Biomechanical stress–strain measurements and thermal shrinkage tests were performed. Results: In the Air Optix Aqua group (30% hydration) without riboflavin film, Young′s modulus and stress at 8% strain were increased significantly versus untreated controls and the effect was 92.4% respectively 86.35% of the standard CXL value. In the SofLens group (59% hydration) without riboflavin film, Young′s modulus and stress at 8% strain were increased significantly versus untreated controls and the effect was 67.04% respectively 65.28% of the standard CXL value. In the Galifa group (72% hydration) without riboflavin film, Young′s modulus and stress at 8% strain were increased significantly versus untreated controls and the effect was about 68.48% respectively 75.52% of the standard CXL value. In all samples with a precorneal riboflavin film under the contact lens, there was no significant biomechanical effect compared to the untreated controls. Similarly, in the hydrothermal experiments at 70°C, there was a typical mushroom pattern with increased resistance to thermal shrinkage in the anterior stroma after standard CXL, a markedly reduced mushroom effect using a riboflavin‐soaked contact lens only and no effect with the use of a riboflavin‐soaked contact lens plus a precorneal riboflavin film. Conclusion: The biomechanical effect of CACXL in porcine corneas is about one‐third less than after standard CXL. The efficacy of CACXL might be improved by reducing or omitting the riboflavin film on the contact lens. Further risk assessment studies are necessary. [ABSTRACT FROM AUTHOR]

Purpose: To compare the antifungal efficacy of corneal collagen cross‐linking with photoactivated riboflavin (PACK‐CXL) and voriconazole in experimental Fusarium solani and Candida albicans keratitis models. Methods: Sixty‐four corneas of 32 New Zealand rabbits were included and divided into two main groups. Intrastromal injection of Fusarium and Candida suspensions was performed, and it was observed that keratitis was formed on the third day. Both groups were randomly separated into the following four groups: control, PACK‐CXL, voriconazole and PACK‐CXL combined with voriconazole. PACK‐CXL was applied using 0.25% riboflavin in an accelerated Dresden protocol (total ultraviolet A dose 5.4 J/cm²). Voriconazole was applied topically as 7x1/day with a dose of 1% (10 mg/ml). Corneal buttons were excised on the tenth day, and microbiological and pathological examinations were performed. Results: The PACK‐CXL and PACK‐CXL combined with voriconazole groups each had 100 colony‐forming unit (CFU/ml) of reproduced micro‐organisms compared with 500 CFU/ml in the voriconazole group and 1500 CFU/ml in the control group (p < 0.001) in the Fusarium keratitis model. The PACK‐CXL combined with voriconazole group had 100 CFU/ml, the PACK‐CXL group had 150 CFU/ml, and the voriconazole group had 200 CFU/ml of reproduced micro‐organisms compared with 4000 CFU/ml in the control group (p < 0.002) in the Candida keratitis model. (p < 0.001). Fewer hyphae and non‐specific stromal changes were observed in the pathological cross sections examined in subgroups that used CXL. Conclusion: There was less fungus reproduction and a lower keratitis score for Fusarium solani and Candida albicans in the treatment groups compared to the control groups, especially in groups that used PACK‐CXL. These results suggest that it is useful to combine PACK‐CXL treatment with medical treatment in the fungal keratitis algorithm at the early stage of the disease. [ABSTRACT FROM AUTHOR]

Abstract The purpose of this study was to measure collagen fiber crimping (CFC) using nonlinear optical imaging of second harmonic generated (SHG) signals to determine the effects of UVA-riboflavin induced corneal collagen crosslinking (UVA CXL) on collagen structure. Two groups, four rabbits each, were treated in the right eye with standard UVA CXL. In vivo confocal microscopy was performed at 1, 2, and 4 weeks after treatment for the first group and up to three months for the second group to measure epithelial/stromal thickness and corneal haze during recovery. Rabbits were sacrificed at one and three months, respectively, and their corneas fixed under pressure. Regions of crosslinking were identified by the presence of collagen autofluorescence (CAF) and then collagen structure was imaged using SHG microscopy. The degree of CFC was determined by measuring the percentage difference between the length of the collagen fiber and the linear distance traveled. CFC was measured in the central anterior and posterior CXL region, the peripheral non-crosslinked region in the same cornea, and the central cornea of the non-crosslinked contralateral eye. No change in corneal thickness was detected after one month, however the stromal thickness surpassed its original baseline thickness at three months by 25.9 μm. Corneal haze peaked at one month and then began to clear. Increased CAF was detected in all CXL corneas, localized to the anterior stroma and extending to 42.4 ± 3.4% and 47.7 ± 7.6% of the corneal thickness at one and three months. There was a significant (P < 0.05) reduction in CFC in the CAF region in all eyes averaging 1.007 ± 0.006 and 1.009 ± 0.005 in one and three month samples compared to 1.017 ± 0.04 and 1.016 ± 0.06 for controls. These results indicate that there is a significant reduction in collagen crimping following UVA CXL of approximately 1%. One possible explanation for this loss of crimping could be shortening of the collagen fibers over the CXL region. Highlights • Corneal thickness remained consistent over time, and corneal haze was shown to peak at one month and then decrease. • CAF showed no decrease between one and three months, indicating persistent mechanical stiffening over time. • Collagen fiber crimping measurements revealed a straighter area of fibers within the crosslinked region, found using CAF. • Cellular staining showed migration of keratocytes into the crosslinked region from the periphery. [ABSTRACT FROM AUTHOR]