July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
Development of Hyaluronic Acid Based Scaffolds to Maintain Stemness in Human Dental Pulp Stem Cell Toward Ocular Regeneration
Author Affiliations & Notes
  • Tao L Lowe
    Pharmaceutical Sciences, Univ of Tennessee Health Science Ctr, Memphis, Tennessee, United States
  • Kumar K Niloy
    Pharmaceutical Sciences, Univ of Tennessee Health Science Ctr, Memphis, Tennessee, United States
  • Muhammad Gulfam
    Pharmaceutical Sciences, Univ of Tennessee Health Science Ctr, Memphis, Tennessee, United States
  • Dong Li
    Department of Bioscience Research, University of Tennessee Health Science Center, Memphis, Tennessee, United States
  • George T._J. Huang
    Department of Bioscience Research, University of Tennessee Health Science Center, Memphis, Tennessee, United States
  • Footnotes
    Commercial Relationships   Tao Lowe, None; Kumar Niloy, None; Muhammad Gulfam, None; Dong Li, None; George Huang, None
  • Footnotes
    Support  NIH R01EY023853
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 3932. doi:
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      Tao L Lowe, Kumar K Niloy, Muhammad Gulfam, Dong Li, George T._J. Huang; Development of Hyaluronic Acid Based Scaffolds to Maintain Stemness in Human Dental Pulp Stem Cell Toward Ocular Regeneration. Invest. Ophthalmol. Vis. Sci. 2019;60(9):3932.

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

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Abstract

Purpose : The self-renewal and multilineage differentiation capability of human dental pulp stem cells (DPSCs) has spurred interest in the application of DPSCs in ocular regeneration. DPSCs need 3D extracellular microenvironment that can provide appropriate physical, chemical, biological and mechanical cues to maintain and promote their pluripotency and differentiation, respectively. The purpose of this work was to evaluate the effect of molecular weight (MW), mechanical property and degradation of 3-D hydrogel scaffolds made of hyaluronic acid (HA) and 2-aminoethyl methacrylate (AEMA) on maintaining the stemness of DPSCs toward regeneration of ocular tissues using DPSCs in the future.

Methods : HA with MW 18 kD or 270 kD was methacrylated by using AEMA. The obtained HA-AEMA macromers were polymerized into hydrogels via UV synthesis. The chemical structures of the macromers were characterized by 1H-NMR. The rheological properties of swollen hydrogels as a function of time were evaluated by using TA ARG2 rheometer. The cytotoxicity of macromers to DPSCs and the survival of DPSCs inside the hydrogels were evaluated by MTT and cell death assays, respectively. The pluripotency of DPSCs grown inside the hydrogels were investigated by assessing the expression of stemness genes SOX2, OCT4 and NANOG using qRT-PCR.

Results : The storage moduli of the HA hydrogels decreased significantly with decreasing the MW of HA and along degradation. HA-AEMA macromers were not toxic to DPSCs and the cells survived in the hydrogels without any cell death for at least 7 days. DPSCs maintained natural spherical shape when the storage moduli of the hydrogels were above 186±44 Pa. The hydrogels increased the expression of stemness genes NANOG and SOX2 at least 4 and 25 folds, respectively, when their storage moduli were 485±59 and 597±67 Pa, respectively.

Conclusions : Hydrogels made of methacrylated HA with 18 kD or 270 kD MW were successfully synthesized and characterized. DPSCs maintained their viability, natural shape and pluripotency in the hydrogels for 7 days. The designed hydrogels with tunable rheological, mechanical and degradation properties have great potential to maintain the native morphology and stemness of DPSCs for ocular regenerative medicine in the future.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

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