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An Inducible Model for Unraveling the Effects of Advanced Glycation End-Products in Collagen with Age and Diabetes

Austin G. Gouldin, View ORCID ProfileJennifer L. Puetzer
doi: https://doi.org/10.1101/2020.09.04.283473
Austin G. Gouldin
1Departments of Biomedical Engineering, Virginia Commonwealth University, Richmond, VA, United States
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Jennifer L. Puetzer
1Departments of Biomedical Engineering, Virginia Commonwealth University, Richmond, VA, United States
2Orthopaedic Surgery, Virginia Commonwealth University, Richmond, VA, United States
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  • ORCID record for Jennifer L. Puetzer
  • For correspondence: jlpuetzer@vcu.edu
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Abstract

In connective tissues there is a clear link between increasing age and degeneration. It is believed advanced glycation end-products (AGEs) play a central role in this degeneration. AGEs are sugar induced non-enzymatic crosslinks which accumulate in collagen with age and diabetes, altering tissue mechanics and cellular function. Despite ample correlative evidence linking collagen glycation to degeneration, little is known how AGEs impact cell-matrix interactions, limiting therapeutic options. One reason for this limited understanding is AGEs are typically induced in vitro using high concentrations of ribose which decrease cell viability and make it impossible to investigate cell-matrix interactions. The objective of this study was to develop a system to trigger AGE accumulation while maintaining cell viability. Using cell-seeded high density collagen gels, we investigated the effect of two different systems for AGE induction, ribose at low concentrations (30, 100, and 200 mM) over 15 days of culture and riboflavin (0.25 mM and 0.75mM) induced with blue light for 40 seconds. We found ribose and riboflavin with blue light are capable of producing a wide range of AGE crosslinks which match and/or exceed reported human AGE levels for various tissues, ages, and diseases, without affecting cell viability and metabolism. Interestingly, a single 40 second treatment of riboflavin and blue light produced similar levels of AGEs as 3 days of 100 mM ribose treatment and matched aged mouse tendon AGE levels. This riboflavin treatment option is an exciting means to trigger AGE crosslinks on demand in vivo or in vitro without impacting cell metabolism or viability and holds great promise for further unraveling the mechanism of AGEs in age and diabetes related tissue degeneration.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted September 04, 2020.
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An Inducible Model for Unraveling the Effects of Advanced Glycation End-Products in Collagen with Age and Diabetes
Austin G. Gouldin, Jennifer L. Puetzer
bioRxiv 2020.09.04.283473; doi: https://doi.org/10.1101/2020.09.04.283473
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An Inducible Model for Unraveling the Effects of Advanced Glycation End-Products in Collagen with Age and Diabetes
Austin G. Gouldin, Jennifer L. Puetzer
bioRxiv 2020.09.04.283473; doi: https://doi.org/10.1101/2020.09.04.283473

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