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Healthcare associated infections (HCAI) represent a significant burden worldwide contributing to morbidity and mortality and result in substantial economic consequences equating to billions annually. Although the impacts of HCAI have been felt for many years, the coronavirus pandemic has had a profound effect, escalating rates of HCAI, even with extensive preventative measures such as vaccination, personal protective equipment, and deep cleaning regimes. Therefore, there is an urgent need for new solutions to mitigate this serious health emergency. In this paper, the fabrication of nitric oxide (NO) releasing dual action polymer coatings for use in healthcare applications is described. The coatings are doped with the NO donor S -nitroso- N -acetylpenicillamine (SNAP) and release high payloads of NO in a sustained manner for in excess of 50 hours. These coatings are extensively characterized in multiple biologically relevant solutions and the antibacterial/antiviral efficacy is studied. For the first time, we assess antibacterial activity in a time course study (1, 2, 4 and 24 h) in both nutrient rich and nutrient poor conditions. Coatings exhibit excellent activity against Pseudomonas aeruginosa and methicillin resistant Staphylococcus aureus (MRSA), with up to complete reduction observed over 24 hours. Additionally, when tested against SARS-CoV-2, the coatings significantly reduced active virus in as little as 10 minutes. These promising results suggest that these coatings could be a valuable addition to existing preventative measures in the fight against HCAIs.

Purpose: To evaluate and compare the biomechanical properties of the eye bank-prepared and surgeon prepared Descemet stripping automated endothelial keratoplasty (DSAEK) tissues., Methods: In this laboratory study, corneal tissues for research were randomly allocated in the following groups: a) surgeon-cut DSAEK and b) eye bank-prepared (pre-cut and pre-loaded) DSAEK. Endothelial cell loss (ECL), immunostaining for tight junction protein ZO-1, elastic modulus, and adhesion force were investigated., Results: ECL was not found to be significantly different between surgeon-cut DSAEK (7.8% ±6.5%), pre-cut DSAEK (8.6% ±2.3%), and pre-loaded DSAEK (11.1% ±4.8%) (P = 0.5910). ZO-1 was expressed equally across all groups. Surgeon-cut DSAEK grafts showed a significantly higher elastic modulus compared to pre-cut and pre-loaded DSAEK groups (P = 0.0047 and P < 0.0001, respectively). Adhesion force was significantly greater in the surgeon-cut DSAEK compared to pre-cut (P < 0.0001) or pre-loaded DSAEK groups (P = 0.0101)., Conclusion: The laboratory data on the biomechanics of DSAEK grafts suggests that surgeon-cut DSAEK grafts present higher elastic modulus and adhesion force compared to eye bank-prepared DSAEK grafts., Competing Interests: None

Aim: To investigate the difference in adhesion and rebubbling rate between eye bank and surgeon prepared Descemet membrane endothelial keratoplasty (DMEK) tissues., Methods: Laboratory and clinical retrospective comparative interventional case series. Research corneal tissues were obtained for laboratory investigation. The clinical study involved patients with endothelial dysfunction who underwent DMEK surgery and tamponade with air. Tissues were stripped using a standard DMEK stripping technique (SCUBA) and shipped as prestripped or loaded in a 2.2 intra-ocular lens cartridge with endothelium facing inwards (preloaded) before transporting from the eye bank to the surgeon. For surgeon prepared tissues, all the grafts were stripped in the theatre and transplanted or stripped in the laboratory and tested immediately. Adhesion force and elastic modulus were measured in the centre and mid-periphery in a laboratory ex vivo investigation using atomic force microscopy, while rebubbling rates were recorded in the clinical study., Results: There was no difference in endothelial cell viability between surgeon or eye bank prepared tissue. Surgeon-stripped DMEK grafts in the laboratory investigation showed significantly higher elastic modulus and adhesion force compared to prestripped and preloaded tissues (p<0.0001). In the clinical data, rebubbling rates of 48%, 40% and 15% were observed in preloaded, prestripped and surgeon-stripped DMEK grafts, respectively. Rebubbling rates were significantly associated with combined cataract surgery (p=0.009) and with time from harvesting the graft to the surgery (p=0.02)., Conclusions: Decreased adhesion forces and elastic modulus in eye bank prepared tissues may contribute to increased rebubbling rates., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2022. Re-use permitted under CC BY. Published by BMJ.)

Purpose: To explore the effect of age on corneal biomechanical properties following corneal cross-linking (CXL)., Methods: A total of 12 pairs of human eye-banked corneas (24 corneas, from 14 females and 10 males) were used in the study. The mean donor age was 48.5 years (ranging from 26 to 71 years). Corneas were divided into three age groups: A (26-41 years), B (42-57 years) and C (58-71 years), with four pairs in each group. For each pair, the right corneas were cross-linked using accelerated CXL with UVA (10 mW/cm 2 ) and riboflavin, while the left corneas served as controls and were not exposed to either UVA irradiation or riboflavin. The corneal elastic modulus of the anterior, mid and posterior corneal stroma was measured using nanoindentation., Results: The difference in the corneal elastic modulus following CXL was significant in the anterior (p = 0.00002) and mid stroma (p = 0.001); however, the difference was not significant in the posterior stroma (p = 0.27) when compared to control corneas. The corneal elastic modulus of the anterior stroma increased by 178.44% in Group A, 119.7% in Group B and 50.73% in Group C compared to control corneas. For the mid stroma, the elastic modulus increased by 47.35% in Group A, 25% in Group B and 24.56% in Group C. No differences were observed in the posterior stroma between age groups., Conclusions: Corneal elasticity showed a greater response to CXL in the younger group compared to older groups. CXL treatment showed effectiveness in enhancing stromal strength, and the effect was concentrated in the anterior and mid stroma with minimal impact on the posterior stroma in all age groups., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Under physiological conditions, the cornea is exposed to various enzymes, some of them have digestive actions, such as amylase and collagenase that may change the ultrastructure (collagen morphology) and sequentially change the mechanical response of the cornea and distort vision, such as in keratoconus. This study investigates the ultrastructure and nanomechanical properties of porcine cornea following incubation with α-amylase and collagenase. Atomic force microscopy (AFM) was used to capture nanoscale topographical details of stromal collagen fibrils (diameter and D-periodicity) and calculate their elastic modulus. Samples were incubated with varying concentrations of α-amylase and collagenase (crude and purified). Dimethylmethylene blue (DMMB) assay was utilised to detect depleted glycosaminoglycans (GAGs) following incubation with amylase. Collagen fibril diameters were decreased following incubation with amylase, but not D-periodicity. Elastic modulus was gradually decreased with enzyme concentration in amylase-treated samples. Elastic modulus, diameter, and D-periodicity were greatly reduced in collagenase-treated samples. The effect of crude collagenase on corneal samples was more pronounced than purified collagenase. Amylase was found to deplete GAGs from the samples. This enzymatic treatment may help in answering some questions related to keratoconus, and possibly be used to build an empirical animal model of keratoconic corneas with different progression levels.

The micromechanical behaviour of the cornea is important for understanding and modelling of many ocular disorders. Whereas inflation testing has been utilised to determine the bulk mechanical properties of the cornea under physiological pressures, micromechanical testing has been limited to unpressurised corneal samples. In this study the micromechanical properties of pressurised porcine corneas were determined using oscillatory nanoindentation coupled with a custom inflation method. Inflation was conducted in phosphate buffered saline (PBS) and tissue culture (TC) solutions. The shear storage modulus (G') and shear loss modulus (G") was determined for corneas inflated corneas with PBS and TC. Central corneal thickness (CCT) was monitored during the inflation (0-60 mmHg). Elastic modulus (E) was also calculated and quantitatively mapped for corneas. The results showed that G' at 15 mmHg was 86.18 ± 16 kPa and 88.86 + 13.54 kPa inflated by PBS and TC respectively. These values increased 3.2 times in an approximate linear relationship to 60 mmHg. G″ at 15 mmHg was 12.5 ± 2.5 kPa and 13.54 + 1.9 kPa inflated by PBS and TC respectively. G″ increased 1.9 times in an approximate linear relationship to 60 mmHg. No significant change was noticed in viscoelastic properties of corneas inflated by TC for 4 h whereas 4 h of hydration on PBS affected the mechanical properties. The central corneal region was found to be stiffer than in peripheral region. Mapping of elasticity revealed a symmetrical distribution of properties that varied with inflation. Our method has potential for measurement of viscoelastic properties of corneas in conditions where there have been localised changes in mechanical properties such as keratoconus., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

There has been a lot of interest in accurately characterising corneal biomechanical properties under intraocular pressure (IOP) to help better understand ocular pathologies that are associated with elevated IOP. This study investigates the novel use of Line-Field Optical Coherence Tomography (LF-OCT) as an elastographic tool for accurately measuring mechanical properties of porcine corneas based on volumetric deformation following varying IOPs. A custom-built LF-OCT was used to measure geometrical and corneal surface displacement changes in porcine corneas under a range of IOPs, from 0-60 mmHg. Corneal thickness, elastic properties and hysteresis were calculated as a function of pressure. In addition, the effects of hydration were explored. We found that the elastic modulus increased in a linear fashion with IOP. Corneal thickness was found to reduce with IOP, decreasing 14% from 0 to 60 mmHg. Prolonged hydration in phosphate buffered saline (PBS) was found to significantly increase the elastic modulus and corneal hysteresis. Our study demonstrates that LF-OCT can be used to accurately measure the elastic properties based on volumetric deformation following physiological pressures. Furthermore, we show that prolonged hydration in PBS has a significant effect on the measured corneal properties.