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Evaluation of Multiphoton Imaging for Analyzing the Nature and Distribution of Advanced Glycation Endproducts (AGE’s) in Diabetic Skin
Suzanne Schwartz, Assistant Professor, Department of Surgery, Weill Medical College of Cornell.
Funding: NIH/NIGMS 1K23GM073728 (4/05-3/10).

The United States is facing an epidemic in type 2 diabetes; of the estimated 21 million people with the disease, approximately one third remain undiagnosed. Early detection and intervention can delay or even prevent the disease; however, current methods for diagnosis are somewhat inaccurate and inconvenient. New methods for accurate diagnosis have been established based on the intrinsic fluorescence characteristics of the abnormally cross-linked and highly insoluble glycation/Maillard reacted proteins termed advanced glycation endproducts (AGE’s) that are characteristic of long-term elevated blood glucose levels (Hull et al., 2004). The objective of this project is to assess the efficacy of multiphoton microscopy for evaluating the distribution of diabetes-associated AGE’s in human skin.

This is a new collaboration. IRB approval for human subject research (1201-027) is now in place.  We propose here to assess MPM for imaging skin AGE’s using biopsies donated from normal and diabetic patients undergoing burn therapy (McCampbell et al., 2002). AGE distribution will be assessed using autofluorescence within the normal skin structure. AGE’s show a peak UV excitation with a broad blue emission, and collagen is thought to be the primary protein that undergoes glycation and glycooxidation (Verzijl et al., 2000). AGE content is known to affect its physical properties, such as its charge, hydrophobicity, turnover and elasticity. Because collagen fibrils are such stable structures, their glycation levels represent long-term glycemia integrators (on the year timescale in skin) with relative insensitivity to short-term fluctuations in glucose levels. Using unstained tissue specimens, we plan to examine whether AGE’s are associated with dermal collagen fibrils and/or capillary-scale blood vessels. Several types of AGE’s have also shown positive correlations with the severity of diabetic neuropathy (Verbeke et al., 1997). To show association with innervation or lack thereof, we will assess AGE autofluorescence as it correlates with Nissl staining in these specimens. MPM identification of AGE’s will also be assessed for possible diagnostic potential. This collaboration is intimately connected to a better understanding the origins of collagen-associated autofluorescence.


Hull, E.L., M.N. Ediger, A.H.T. Unione, E.K. Deemer, M.L. Stroman, and J.W. Baynes (2004) Noninvasive, optical detection of diabetes: model studies with porcine skin. Opt Express 12(19):4496-4510.

McCampbell, B., N. Wasif, A. Rabbitts, L. Staiano-Coico, R.W. Yurt, and S. Schwartz (2002) Diabetes and burns: retrospective cohort study. J Burn Care Rehabil 23(3):157-166.

Verzijl, N., J. DeGroot, S.R. Thorpe, R.A. Bank, J.N. Shaw, T.J. Lyons, J.W. Bijlsma, F.P. Lafeber, J.W. Baynes, and J.M. TeKoppele (2000) Effect of collagen turnover on the accumulation of advanced glycation end products. J Biol Chem 275(50):39027-39031.

Verbeke, P., M. Perichon, C. Borot-Laloi, J. Schaeverbeke, and H. Bakala (1997) Accumulation of advanced glycation endproducts in the rat nephron: link with circulating AGEs during aging. J Histochem Cytochem 45(8):1059-1068.




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