We present a technique for estimating the density of the human macular pigment noninvasively that takes advantage of the autofluorescence of lipofuscin, which is normally present in the human retinal pigment epithelium. By measuring the intensity of fluorescence at 710 nm, where macular pigment has essentially zero absorption, and stimulating the fluorescence with two wavelengths, one well absorbed by macular pigment and the other minimally absorbed by macular pigment, we can make accurate single-pass measurements of the macular pigment density. We used the technique to measure macular pigment density in a group of 159 subjects with normal retinal status ranging in age between 15 and 80 years. Average macular pigment density was 0.48 +/- 0.16 density unit (D.U.) for a 2 degrees -diameter test field. We show that these estimates are highly correlated with reflectometric (mean: 0.23 +/- 0.07 D.U.) and psychophysical (mean: 0.37 +/- 0.26 D.U.; obtained by heterochromatic flicker photometry) estimates of macular pigment in the same subjects, despite the fact that systematic differences in the estimated density exist between techniques. Repeat measurements over both short- and long-time intervals indicate that the autofluorescence technique is reproducible: The mean absolute difference between estimates was less than 0.05 D.U., superior to the reproducibility obtained by reflectometry and flicker photometry. To understand the systematic differences between density estimates obtained from the different methods, we analyzed the underlying assumptions of each technique. Specifically, we looked at the effect of self-screening by visual pigment, the effect of changes in optical property of the deeper retinal layers, including the role of retinal pigmented epithelium melanin, and the role of secondary fluorophores and reflectors in the anterior layers of the retina.

Individual variations in the spatial profile of macular pigment (MP) density were measured for 32 subjects. Peak density of MP measured with a 460-nm, 12-arcmin stimulus averaged 0.58, standard deviation (SD) = 0.26, with a range of 0.175 to 1.39. To assess the symmetry of the MP distribution, MP density was measured on the horizontal and vertical meridians at +/-1 deg eccentricity. The density varied by no more than 16% at these four locations, indicating a basically symmetric distribution. Based on a linear interpolation between measured locations, the width of the spatial distribution of MP at half the maximal density averaged 1.03 deg, SD = 0.38, with a range of 0.25 to 1.9 deg. The average spatial profile of MP density across subjects was fitted with both an exponential and a Gaussian function. An exponential decay with eccentricity explained more variance in the data than did a Gaussian function. Assuming an exponential decay with eccentricity, once MP density has been measured in the center of the retina (denoted A), MP density at more eccentric locations (X, deg) can be predicted with a standardized equation (MP = A x 10(-0.42x)). For individual cases, small deviations from an exponential function suggest the existence of minor flanking peaks or shoulders for 40% of the subjects. We also examined the temporal stability of the MP profile of four subjects over a time span of 4-14 months and for a single spatial location for ten subjects over a time span of 1-16 years. These longitudinal data show that differences in MP density among subjects are maintained over time, if dietary patterns are stable.

Background: Lutein and zeaxanthin are the only carotenoids in the macular region of the retina (referred to as macular pigment [MP]). Foods that are rich in lutein and zeaxanthin can increase MP density. Response to dietary lutein and zeaxanthin in other tissues has not been studied. Objective: The objective of this study was to examine tissue responses to dietary lutein and zeaxanthin and relations among tissues in lutein and zeaxanthin concentrations. Design: Seven subjects consumed spinach and corn, which contain lutein and zeaxanthin, with their daily diets for 15 wk. At 0, 4, 8, and 15 wk and 2 mo after the study, serum, buccal mucosa cells, and adipose tissue were analyzed for carotenoids, and MP density was measured. Results: Serum and buccal cell concentrations of lutein increased significantly from baseline during dietary modification. Serum zeaxanthin concentrations were greater than at baseline only at 4 wk, whereas buccal cell and adipose tissue concentrations of zeaxanthin did not change. Adipose tissue lutein concentrations peaked at 8 wk. Changes in adipose tissue lutein concentration were inversely related to the changes in MP density, suggesting an interaction between adipose tissue and retina in lutein metabolism. To investigate the possibility of tissue interactions, we examined cross-sectional relations among serum, tissue, and dietary lutein concentrations, anthropometric measures, and MP density in healthy adults. Significant negative correlations were found between adipose tissue lutein concentrations and MP for women, but a significant positive relation was found for men. Conclusion: Sex differences in lutein metabolism may be an important factor in tissue interactions and in determining MP density.Am J Clin Nutr 2000;71:1555-62.

MP is strongly related to improvements in glare disability and photostress recovery in a manner strongly consistent with its spectral absorption and spatial profile. Four to 6 months of 12 mg daily L + Z supplementation significantly increases MPOD and improves visual performance in glare for most subjects.

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Background: High levels of xanthophyll carotenoids lutein (L) and zeaxanthin (Z) in the central nervous system have been previously correlated with improved cognitive function in community-dwelling older adults. In this study, we tested the effects of supplementing L and Z on older men and women with a range of baseline cognitive abilities.Objective: The purpose of this study was to determine whether or not supplementation with L+Z could improve cognitive function in community-dwelling, older adults.Design: Double-masked, randomized, placebo-controlled trial. A total of 62 older adults were randomized into groups receiving either 12 mg L+Z or a visually identical placebo. Data from 51 participants (M = 73.7 years) were available for analysis. Retinal L+Z levels (macular pigment optical density, MPOD) were measured psychophysically using heterochromatic flicker photometry as a biomarker of cortical L+Z levels. Cognitive function was measured using the CNS Vital Signs computerized test platform.Results: Participants receiving the active L+Z supplement had statistically significant increases in MPOD (p < 0.03) and improvements in complex attention (p < 0.02) and cognitive flexibility domains (p < 0.04), relative to participants taking the placebo. A trend was also seen for the executive function domain (p = 0.073). In male participants only, supplementation yielded improved composite memory (p = 0.04).Conclusions: Supplementation with L+Z improved cognitive function in community-dwelling, older men and women.

MP is strongly related to improvements in glare disability and photostress recovery in a manner consistent with its spectral absorption and spatial profile.

Retinal carotenoids can be measured in epidemiologic studies. In this study, MPOD was associated with lutein + zeaxanthin in the diet and the serum. Retinal concentrations, however, were influenced by other factors as well. To understand the effect of dietary lutein + zeaxanthin intake on the retina and risk of age-related eye disease, future studies should include measures of macular concentrations of these pigments.