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    Selective two-photon collagen crosslinking: Why more is better

    Technique mechanically strengthens corneal tissue in situ without damage to surrounding untreated tissue


    In this animal study, they tested the ability to control the technology’s spatial selectivity in tissue and pinpoint its clinical effects more precisely. They used femtosecond laser pulses and riboflavin for 2P-CXL in the corneas of the animals.

    Following treatment, confocal Brillouin microscopy was performed to validate their results. Brillouin microscopy is a technique developed recently by Dr. Yun and colleagues that uses light to probe mechanical properties non-invasively in cells and tissues.

    Related: Predicting, treating keratoconus in 2016

    Currently, Brillouin microscopy is used to study the cornea in normal subjects and patients with keratoconus.

    “While conventional CXL crosslinks the entire stromal depth of the cornea (over 500 μm), 2P-CXL enables crosslinking of any arbitrary three-dimensional region within the cornea,” explained Sheldon Kwok, BA, lead author on the paper. Kwok is an MD/PhD candidate, Harvard-Massachusetts Institute of Technology Health Sciences and Technology, Harvard Medical School, and a member of the Yun Bio-Optics Lab, Wellman Center for Photomedicine, Massachusetts General Hospital, Boston.

    Recent: When is crosslinking appropriate?

    The investigators used their technique to demonstrate crosslinking of a rectangular region 50 μm below the cornea surface. Using Brillouin microscopy, they found the crosslinked region was significantly stiffer than surrounding regions.

    Importantly, the tissue stiffening induced by 2P-CXL was found to be comparable to that achieved with conventional CXL.

    More: Addressing challenges of Acanthamoeba keratitis

    The investigators concluded, “…we have demonstrated three-dimensional selective CXL in corneal tissue. Our technique involves in situ 2P-CXL of an arbitrary three-dimensional structure, and verification and characterization with confocal Brillouin microscopy.”

    Compare with conventional procedures

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