June 2023
Volume 64, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2023
Characterization of the molecular weight of hyaluronan in eye products using a novel method of size exclusion high-pressure liquid chromatography
Author Affiliations & Notes
  • Sudhir Verma
    College of Optometry, University of Houston, Houston, Texas, United States
    Deen Dayal Upadhyaya College, University of Delhi, New Delhi, Delhi, India
  • Lawrence Nguyen
    College of Optometry, University of Houston, Houston, Texas, United States
  • Xiao Lin
    College of Optometry, University of Houston, Houston, Texas, United States
  • Sudan Puri
    College of Optometry, University of Houston, Houston, Texas, United States
  • Vincent Hascall
    Cleveland Clinic, Cleveland, Ohio, United States
  • Tarsis G Ferreira
    College of Optometry, University of Houston, Houston, Texas, United States
  • Vivien Jane Coulson-Thomas
    College of Optometry, University of Houston, Houston, Texas, United States
  • Footnotes
    Commercial Relationships   Sudhir Verma None; Lawrence Nguyen None; Xiao Lin None; Sudan Puri None; Vincent Hascall None; Tarsis Ferreira None; Vivien Coulson-Thomas None
  • Footnotes
    Support  NIH/ NEI grant R01 EY029289 and R01 EY033024; Core grant P30 EY07551
Investigative Ophthalmology & Visual Science June 2023, Vol.64, 4487. doi:
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      Sudhir Verma, Lawrence Nguyen, Xiao Lin, Sudan Puri, Vincent Hascall, Tarsis G Ferreira, Vivien Jane Coulson-Thomas; Characterization of the molecular weight of hyaluronan in eye products using a novel method of size exclusion high-pressure liquid chromatography. Invest. Ophthalmol. Vis. Sci. 2023;64(8):4487.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Hyaluronan (HA) exists in two forms, high molecular weight HA (HMWHA) and low molecular weight HA (LMWHA), which have distinct physiological functions. Therefore, it is imperative to know the form of HA within pharmaceutical products, including eye products. This study developed an accurate, sensitive, and quantitative method to characterize the form of HA in eye products. Thereafter, the effects of the HA-containing eye products on corneal wound healing were investigated.

Methods : The MW distributions and concentrations of HA in over-the-counter eye products were determined by size exclusion chromatography (SEC) high pressure liquid chromatography (HPLC). The effects of the eye products containing HA on corneal wound healing were characterized both in vitro and in vivo using the scratch assay and the debridement wound model, respectively.

Results : The concentrations and MWs of HA were successfully determined within a range of 0.014 to 0.25 mg/mL using SEC HPLC. The concentrations of HA in the ophthalmic products varied from 0.14 to 4.0 mg/mL and the MWs varied from ~100 kDa to 2500 kDa. All but one HA-containing eye product had an inhibitory effect on corneal wound healing, while pure HA promoted corneal wound healing.

Conclusions : A novel SEC-HPLC method was developed for quantifying and characterizing the MW of HA in eye products. Although HA promoted corneal wound healing, HA-containing eye products inhibited corneal wound healing, likely caused by preservatives.

This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.

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