Nanotopography by chromatic confocal microscopy of the endothelium in Fuchs endothelial corneal dystrophy, pseudophakic bullous keratopathy and healthy corneas.

AIM: To investigate the interest of chromatic confocal microscopy (CCM) to characterise guttae in Fuchs endothelial corneal dystrophy (FECD). METHODS: Descemet's membranes (DM) were obtained during endothelial keratoplasty in patients with FECD and pseudophakic bullous keratopathy (PBK). They were compared with healthy samples obtained from body donation to science. Samples were fixed in 0.5% paraformaldehyde and flat mounted. Surface roughness of DMs was quantified using CCM and the AltiMap software that provided the maximum peak (Sp) and valley (Sv) heights, the mean square roughness (Rq) and the asymmetry coefficient (Ssk). RESULTS: The physiological roughness of healthy samples was characterised by an Rq of 0.12±0.05 µm, which was two times rougher than in PBK (Rq=0.06±0.03 µm), but both were still flat with a symmetrical distribution between peaks and valleys (Ssk close to 0, npeaks=nvalleys), smaller than 1 µm. In FECD, the maximum peak height was 5.10±2.40 µm, up to 5.8 and 8.3 times higher than the control and PBK, respectively. The maximum valley depth was half than the peak (2.28±0.89 µm). The surface with guttae was very rough (Rq=0.45±0.14 µm) and the Ssk=1.84± 0.43 µm, greater than 0, confirms an asymmetric surface with high peaks and low valleys (npeaks>nvalleys). Moreover, the CCM provided quantitative parameters allowing to distinguish different types of guttae from different patients. CONCLUSIONS: CCM is an innovative approach to describe and quantify different morphologies of guttae. It could be useful to analyse the different stages of FECD and define subgroups of patients.

Ultrafast Laser Patterning of Metals Commonly Used in Medical Industry: Surface Roughness Control with Energy Gradient Pulse Sequences.

Ultrafast laser ablation is widely used as a versatile method for accurate micro-machining of polymers, glasses and metals for a variety of industrial and biomedical applications. We report on the use of a novel process parameter, the modulation of the laser pulse energy during the multi-scan texturing of surfaces. We show that this new and straightforward control method allows us to attain higher and lower roughness (Ra) values than the conventional constant pulse energy irradiation sequence. This new multi-scanning laser ablation strategy was conducted on metals that are commonly used in the biomedical industry, such as stainless steel, titanium, brass and silver samples, using a linear (increasing or decreasing) gradient of pulse energy, i.e., varying the pulse energy across successive laser scans. The effects of ablation were studied in terms of roughness, developed interfacial area ratio, skewness and ablation efficiency of the processed surfaces. Significantly, the investigation has shown a global trend for all samples that the roughness is minimum when a decreasing energy pulse sequence is employed, i.e., the irradiation sequence ends up with the applied laser fluences close to threshold laser fluences and is maximum with increasing energy distribution. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) analysis on single craters with the three different energy deposition conditions revealed a chaotic and random material redistribution in the cases of uniform and increasing energy distributions and the presence of regular laser-induced periodic surface structures (LIPSS) at the bottom of the ablation region in the case of decreasing energy distribution. It is also shown that the ablation efficiency of the ablated surfaces does not significantly change between the three cases. Therefore, this novel energy control strategy permits the control of the roughness of the processed surfaces without losing the ablation efficiency.

Effect of Texturing Environment on Wetting of Biomimetic Superhydrophobic Surfaces Designed by Femtosecond Laser Texturing.

Inspired by Euphorbia leaves, micrometric pillars are designed on 316L stainless steel surfaces using a femtosecond laser to achieve superhydrophobicity. In this study, we focus on wetting behavior evolution as a function of time and chemical environment. Two types of texturing designs are performed: the laser texturing of micrometric square pillars, and the laser texturing of micrometric square pillars whose tops were irradiated using various fluences to obtain a different topography on the nanometric scale. Two laser texturing environments are considered in both cases: a CO2 flow and ambient air. The main result is that 250 days after laser texturing, steady-state contact angles (SSCA) were above 130° no matter what the environment was. We also study the effect of regular wetting over time. Comparing the results of surfaces for which wetting over time was conducted and that of the undisturbed surfaces for 250 days demonstrates that performing wetting measurements when the surface is not stable led to major changes in droplet behavior. Our surfaces have a unique wettability in which droplets are in an intermediate state. Finally, using a CO2 flow did not help reach higher SSCA, but it limited the effect of regular wetting measurements.

Additive and Substractive Surface Structuring by Femtosecond Laser Induced Material Ejection and Redistribution.

A novel additive surface structuring process is devised, which involves localized, intense femtosecond laser irradiation. The irradiation induces a phase explosion of the material being irradiated, and a subsequent ejection of the ablative species that are used as additive building blocks. The ejected species are deposited and accumulated in the vicinity of the ablation site. This redistribution of the material can be repeated and controlled by raster scanning and multiple pulse irradiation. The deposition and accumulation cause the formation of µm-scale three-dimensional structures that surpass the initial surface level. The above-mentioned ablation, deposition, and accumulation all together constitute the proposed additive surface structuring process. In addition, the geometry of the three-dimensional structures can be further modified, if desirable, by a subsequent substractive ablation process. Microstructural analysis reveals a quasi-seamless conjugation between the surface where the structures grow and the structures additively grown by this method, and hence indicates the mechanic robustness of these structures. As a proof of concept, a sub-mm sized re-entrant structure and pillars are fabricated on aluminum substrate by this method. Single units as well as arrayed structures with arbitrary pattern lattice geometry are easily implemented in this additive surface structuring scheme. Engineered surface with desired functionalities can be realized by using this means, i.e., a surface with arrayed pillars being rendered with superhydrophobicity.