Fluorescence lifetime imaging ophthalmoscopy in type 2 diabetic patients who have no signs of diabetic retinopathy

Abstract. The time-resolved autofluorescence of the eye is used for the detection of metabolic alteration in diabetic patients who have no signs of diabetic retinopathy. One eye from 37 phakic and 11 pseudophakic patients with type 2 diabetes, and one eye from 25 phakic and 23 pseudophakic healthy subjects were included in the study. After a three-exponential fit of the decay of autofluorescence, histograms of lifetimes τi, amplitudes αi, and relative contributions Qi were statistically compared between corresponding groups in two spectral channels (490<ch1<560  nm, 560<ch2<700  nm). The change in single fluorophores was estimated by applying the Holm–Bonferroni method and by calculating differences in the sum histograms of lifetimes. Median and mean of the histograms of τ2, τ3, and α3 in ch1 show the greatest differences between phakic diabetic patients and age-matched controls (p<0.000004). The lack of pixels with a τ2 of ∼360  ps, the increased number of pixels with τ2>450  ps, and the shift of τ3 from ∼3000 to 3700 ps in ch1 of diabetic patients when compared with healthy subjects indicate an increased production of free flavin adenine dinucleotide, accumulation of advanced glycation end products (AGE), and, probably, a change from free to protein-bound reduced nicotinamide adenine dinucleotide at the fundus. AGE also accumulated in the crystalline lens.

[1]  M. Sonka,et al.  Early neurodegeneration in the retina of type 2 diabetic patients. , 2012, Investigative ophthalmology & visual science.

[2]  Larry D Hubbard,et al.  Evaluation of the age-related eye disease study clinical lens grading system AREDS report No. 31. , 2010, Ophthalmology.

[3]  R. Holman,et al.  Intensive blood-glucose control with sulphonylureas or insulin compared with conventional treatment and risk of complications in patients with type 2 diabetes (UKPDS 33). UK Prospective Diabetes Study (UKPDS) Group. , 1998 .

[4]  Y. Khalifa,et al.  Neuroprotective and blood-retinal barrier-preserving effects of cannabidiol in experimental diabetes. , 2005, The American journal of pathology.

[5]  S. Genuth,et al.  The effect of intensive treatment of diabetes on the development and progression of long-term complications in insulin-dependent diabetes mellitus. , 1993, The New England journal of medicine.

[6]  Martin Hammer,et al.  Spectral and time-resolved studies on ocular structures , 2007, European Conference on Biomedical Optics.

[7]  D. Schweitzer,et al.  Repeatability of Autofluorescence Lifetime Imaging at the Human Fundus in Healthy Volunteers , 2013, Current eye research.

[8]  S. Holm A Simple Sequentially Rejective Multiple Test Procedure , 1979 .

[9]  Qian Rong-l,et al.  A better comprehension of “Management of Hyperglycemia in Type 2 Diabetes:A Patient-Centered Approach—Position Statement of the American Diabetes Association (ADA) and the European Association for the Study of Diabetes (EASD)” , 2013 .

[10]  Norman Fleischer,et al.  The effect of intensive treatment of diabetes on the development and progression of long-term complications in insulin-dependent diabetes mellitus. The Diabetes Control and Complications Trial Research Group. , 1993 .

[11]  D. Schweitzer,et al.  [Limits of the confocal laser-scanning technique in measurements of time-resolved autofluorescence of the ocular fundus]. , 2005, Biomedizinische Technik. Biomedical engineering.

[12]  R. Kowluru,et al.  Diabetes-induced mitochondrial dysfunction in the retina. , 2003, Investigative ophthalmology & visual science.

[13]  C K Dorey,et al.  In vivo fluorescence of the ocular fundus exhibits retinal pigment epithelium lipofuscin characteristics. , 1995, Investigative ophthalmology & visual science.

[14]  Ji Ho Yang,et al.  Retinal Neurodegeneration in Type II Diabetic Otsuka Long-Evans Tokushima Fatty Rats. , 2013, Investigative ophthalmology & visual science.

[15]  Ammasi Periasamy,et al.  Investigation of tryptophan–NADH interactions in live human cells using three-photon fluorescence lifetime imaging and Förster resonance energy transfer microscopy , 2013, Journal of biomedical optics.

[16]  T. Gardner,et al.  Retinal neurodegeneration: early pathology in diabetes , 2000, Clinical & experimental ophthalmology.

[17]  B. Berkowitz,et al.  Reexamining the hyperglycemic pseudohypoxia hypothesis of diabetic oculopathy. , 2006, Investigative ophthalmology & visual science.

[18]  D. Puro,et al.  Rapid, noninvasive detection of diabetes-induced retinal metabolic stress. , 2008, Archives of ophthalmology.

[19]  D. Schweitzer,et al.  Detection of early metabolic alterations in the ocular fundus of diabetic patients by time-resolved autofluorescence of endogenous fluorophores , 2011, European Conference on Biomedical Optics.

[20]  Grading diabetic retinopathy from stereoscopic color fundus photographs--an extension of the modified Airlie House classification. ETDRS report number 10. Early Treatment Diabetic Retinopathy Study Research Group. , 1991, Ophthalmology.

[21]  D. Schweitzer,et al.  Time-resolved autofluorescence imaging of human donor retina tissue from donors with significant extramacular drusen. , 2012, Investigative ophthalmology & visual science.

[22]  J. Nyengaard,et al.  Early neural and vascular dysfunctions in diabetic rats are largely sequelae of increased sorbitol oxidation. , 2010, Antioxidants & redox signaling.

[23]  J. S. McCasland,et al.  Metabolic Mapping , 2000, Current protocols in neuroscience.

[24]  W. Webb,et al.  Conformational Dependence of Intracellular NADH on Metabolic State Revealed by Associated Fluorescence Anisotropy*♦ , 2005, Journal of Biological Chemistry.

[25]  E. Araki,et al.  Intensive insulin therapy prevents the progression of diabetic microvascular complications in Japanese patients with non-insulin-dependent diabetes mellitus: a randomized prospective 6-year study. , 1995, Diabetes research and clinical practice.

[26]  N. Ramanujam,et al.  In vivo multiphoton microscopy of NADH and FAD redox states, fluorescence lifetimes, and cellular morphology in precancerous epithelia , 2007, Proceedings of the National Academy of Sciences.

[27]  D. Schweitzer,et al.  Towards metabolic mapping of the human retina , 2007, Microscopy research and technique.

[28]  Gwénolé Quellec,et al.  Quantitative analysis of fluorescence lifetime measurements of the macula using the fluorescence lifetime imaging ophthalmoscope in healthy subjects. , 2014, Investigative ophthalmology & visual science.

[29]  T. Gardner,et al.  Neural apoptosis in the retina during experimental and human diabetes. Early onset and effect of insulin. , 1998, The Journal of clinical investigation.

[30]  Uk-Prospective-Diabetes-Study-Group Intensive blood-glucose control with sulphonylureas or insulin compared with conventional treatment and risk of complications in patients with type 2 diabetes (UKPDS 33) , 1998, The Lancet.

[31]  Dietrich Schweitzer,et al.  Two-photon excited fluorescence microscopy application for ex vivo investigation of ocular fundus samples , 2011, European Conference on Biomedical Optics.

[32]  T. Gardner,et al.  Analysis of glucose metabolism in diabetic rat retinas. , 2006, American journal of physiology. Endocrinology and metabolism.

[33]  D. Schweitzer Ophthalmic applications of FLIM , 2014 .

[34]  T. Gardner,et al.  The significance of vascular and neural apoptosis to the pathology of diabetic retinopathy. , 2011, Investigative ophthalmology & visual science.