Incisional surface quality of electron-beam irradiated cornea-extracted lenticule for stromal keratophakia: high nJ-energy vs. low nJ-energy femtosecond laser.

Introduction: Corneal lenticules can be utilized as an additive material for stromal keratophakia. However, following extraction, they must be reimplanted almost immediately or cryopreserved in lenticule banks. Electron-beam (E-beam) irradiated corneas permit room-temperature storage for up to 2 years, enabling keratophakia to be performed on demand. This study aims to compare the performance of high nano Joule (nJ)-energy (VisuMax) and low nJ-energy (FEMTO LDV) femtosecond laser systems on the thickness consistency and surface quality and collagen morphology of lenticules produced from fresh and E-beamed corneas. Methods: A total of 24 lenticules with -6.00 dioptre power were cut in fresh human donor corneas and E-beamed corneas with VisuMax and FEMTO LDV. Before extraction, the thickness of the lenticules was measured with anterior segment-optical coherence tomography (AS-OCT). The incisional surface roughness of extracted lenticules was analyzed using atomic force microscopy (AFM) and scanning electron microscopy (SEM). Multiphoton microscopy was then used to assess the surface collagen morphometry. Results: The E-beamed lenticules that were cut using FEMTO LDV were significantly thicker than the fresh specimens as opposed to those created with VisuMax, which had a similar thickness as the fresh lenticules. On the vertex, they were ∼11% thicker than the fresh lenticules. The surface roughness (Rq) of E-beamed lenticules incised with FEMTO LDV did not differ significantly from the fresh lenticules. This contrasted with the VisuMax-fashioned lenticules, which showed notably smoother surfaces (∼36 and ∼20% lower Rq on anterior and posterior surfaces, respectively) on the E-beamed than the fresh lenticules. The FEMTO LDV induced less cumulative changes to the collagen morphology on the surfaces of both fresh and E-beamed lenticules than the VisuMax. Conclusion: It has been previously demonstrated that the low nJ-energy FEMTO LDV produced a smoother cutting surface compared to high nJ-energy VisuMax in fresh lenticules. Here, we showed that this effect was also seen in the E-beamed lenticules. In addition, lower laser energy conferred fewer changes to the lenticular surface collagen morphology. The smaller disparity in surface cutting quality and collagen disturbances on the E-beamed lenticules could be beneficial for the early visual recovery of patients who undergo stromal keratophakia.

Electron beam-irradiated donor cornea for on-demand lenticule implantation to treat corneal diseases and refractive error.

The cornea is the major contributor to the refractive power of the eye, and corneal diseases are a leading cause of reversible blindness. The main treatment for advanced corneal disease is keratoplasty: allograft transplantation of the cornea. Examples include lenticule implantation to treat corneal disorders (e.g. keratoconus) or correct refractive errors. These procedures are limited by the shelf-life of the corneal tissue, which must be discarded within 2-4 weeks. Electron-beam irradiation is an emerging sterilisation technique, which extends this shelf life to 2 years. Here, we produced lenticules from fresh and electron-beam (E-beam) irradiated corneas to establish a new source of tissue for lenticule implantation. In vitro, in vivo, and ex vivo experiments were conducted to compare fresh and E-beam-irradiated lenticules. Results were similar in terms of cutting accuracy, ultrastructure, optical transparency, ease of extraction and transplantation, resilience to mechanical handling, biocompatibility, and post-transplant wound healing process. Two main differences were noted. First, ∼59% reduction of glycosaminoglycans resulted in greater compression of E-beam-irradiated lenticules post-transplant, likely due to reduced corneal hydration-this appeared to affect keratometry after implantation. Cutting a thicker lenticule would be required to ameliorate the difference in refraction. Second, E-beam-sterilised lenticules exhibited lower Young's modulus which may indicate greater care with handling, although no damage or perforation was caused in our procedures. In summary, E-beam-irradiated corneas are a viable source of tissue for stromal lenticules, and may facilitate on-demand lenticule implantation to treat a wide range of corneal diseases. Our study suggested that its applications in human patients are warranted. STATEMENT OF SIGNIFICANCE: Corneal blindness affects over six million patients worldwide. For patients requiring corneal transplantation, current cadaver-based procedures are limited by the short shelf-life of donor tissue. Electron-beam (E-beam) sterilisation extends this shelf-life from weeks to years but there are few published studies of its use. We demonstrated that E-beam-irradiated corneas are a viable source of lenticules for implantation. We conducted in vitro, in vivo, and ex vivo comparisons of E-beam and fresh corneal lenticules. The only differences exhibited by E-beam-treated lenticules were reduced expression of glycosaminoglycans, resulting in greater tissue compression and lower refraction suggesting that a thicker cut is required to achieve the same optical and refractive outcome; and lower Young's modulus indicating extra care with handling.

Effects of temperature and fluid media on the scroll width size of the Descemet's membrane endothelial keratoplasty (DMEK) donor graft.

AIM: Our study was conducted to evaluate whether higher temperature leads to increased - or wider - scroll widths of the Descemet's membrane endothelial keratoplasty (DMEK) donor graft. PURPOSE: To investigate the effects of temperature and fluid media on the DMEK donor graft scroll widths. MATERIALS AND METHODS: This research work was a laboratory investigation. Baseline cell count was taken via specular microscopy for the donor corneas at room temperature (20°C-25°C). The endothelium sides of the donor corneas were stained with Trypan Blue Solution 0.4% for 30 s, and the Descemet's membranes were stripped. The DMEK donor grafts were placed into three different fluid media - Optisol®, Balanced Salt Solution (BSS), and BSS PLUS® (BSS Plus). The DMEK donor grafts were then transferred into cold temperature (4°C) for 60 min, after which the donor grafts' scroll widths were examined and measured. The donor grafts were then warmed in the incubator and brought to physiological temperature (35°C-37°C), and their scroll widths were examined and measured again. RESULTS: In 30 measurements of ten tissues across three temperature and fluid conditions, the average scroll width measured 1.73 mm, ranging from 1.1 to 2.9 mm. In a mixed linear model, the scroll widths increased with temperature (P=0.02). There was no significant difference in scroll widths among the three solutions (P=0.84, mixed linear model). CONCLUSION: We observed an increase in DMEK donor graft scroll widths with higher temperatures. The usage of BSS Plus as media solution could also lead to smaller DMEK donor graft scroll widths, compared with BSS, but our study does not establish this.

Enhanced bioactivity and osseointegration of PEEK with accelerated neutral atom beam technology.

Polyetheretherketone (PEEK) is growing in popularity for orthopedic, spinal, and trauma applications but has potential significant limitations in use. PEEK is biocompatible, similar in elasticity to bone, and radiolucent, but is inert and therefore does not integrate well with bone. Current efforts are focusing on increasing the bioactivity of PEEK with surface modifications to improve the bone-implant interface. We used a novel Accelerated Neutral Atom Beam (ANAB) technology to enhance the bioactivity of PEEK. Human osteoblast-like cells seeded on ANAB-treated PEEK result in significantly enhanced proliferation compared with control PEEK. Cells grown on ANAB-treated PEEK increase osteogenic expression of ALPL (1.98-fold, p < 0.002), RUNX2 (3.20-fold, p < 0.002), COL1A (1.94-fold, p < 0.015), IBSP (2.78-fold, p < 0.003), and BMP2 (1.89-fold, p < 0.004). Cells grown on these treated surfaces also lead to an increased mineralization (6.4-fold at 21 days, p < 0.0005). In an ovine study, ANAB-treated PEEK implants resulted in enhanced bone-in-contact by 3.09-fold (p < 0.014), increased push-out strength (control 1959 ± 1445 kPa; ANAB 4068 ± 1197 kPa, p < 0.05), and evidence of bone ingrowth at both the early (4 weeks) and later (12 weeks) time points. Taken together, these data suggest that ANAB treatment of PEEK has the potential to enhance its bioactivity, leading to bone formation and significantly decreasing osseointegration time of orthopedic and spinal implants. ANAB treatment, therefore, may significantly enhance the performance of PEEK medical implants and lead to improved clinical outcomes. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 531-543, 2017.