Characterization of the Molecular Weight of Hyaluronan in Eye Products Using a Novel Method of Size Exclusion High-Pressure Liquid Chromatography

Nguyen, Lawrence; Lin, Xiao-Biao; Verma, Sudhir Kumar; Puri, Sudan; Hascall, Vincent C.; Gesteira, Tarsis F.; Coulson‐Thomas, Vivien Jane

doi:10.1167/tvst.12.4.13

Cited by 7 publications

(2 citation statements)

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“…Size exclusion chromatography (SEC) was used to characterize the molecular weight of HA, as previously shown [ 64 ]. For such, two Advanced Bio SEC 300 Å 4.6 × 300 mm columns (Agilent) and one Zorbax GF-250 4.6 × 250 mm column (Agilent) were used in tandem with the UltiMate™ 3000 Dionex system (Thermo Fischer Scientific, Waltham, MA, USA).…”

Section: Methodsmentioning

confidence: 99%

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ROS-Mediated Fragmentation Alters the Effects of Hyaluronan on Corneal Epithelial Wound Healing

Lin,

Moreno,

Nguyen

et al. 2023

Biomolecules

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A buildup of reactive oxygen species (ROS) occurs in virtually all pathological conditions. Hyaluronan (HA) is a major extracellular matrix component and is susceptible to oxidation by reactive oxygen species (ROS), yet the precise chemical structures of oxidized HA products (oxHA) and their physiological properties remain largely unknown. This study characterized the molecular weight (MW), structures, and physiological properties of oxHA. For this, high-molecular-weight HA (HMWHA) was oxidized using increasing molar ratios of hydrogen peroxide (H2O2) or hypochlorous acid (HOCl). ROS lead to the fragmentation of HA, with the oxHA products produced by HOCl exhibiting an altered chemical structure while those produced by H2O2 do not. HMWHA promotes the viability of human corneal epithelial cells (hTCEpi), while low MWHA (LMWHA), ultra-LMWHA (ULMWHA), and most forms of oxHA do not. HMWHA and LMWHA promote hTCEpi proliferation, while ULMWHA and all forms of oxHA do not. LMWHA and some forms of oxHA promote hTCEpi migration, while HMWHA does not. Finally, all native forms of HA and oxHA produced by HOCl promote in vivo corneal wound healing, while oxHA produced by H2O2 does not. Taken together, our results show that HA fragmentation by ROS can alter the physiological activity of HA by altering its MW and structure.

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Section: Methodsmentioning

confidence: 99%

“…* represents p ≤ 0.05. Images for PBS and HMWHA in panel B, D, and F are adapted from [ 64 ], and copyright has been gained from the authors and the journal TVST.…”

Section: Figurementioning

confidence: 99%