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BACKGROUND: Vitreous opacities and posterior vitreous detachment (PVD) disturb vision by degrading contrast sensitivity (AJO 172:7-12, 2016). Increased light scattering is the presumed mechanism. To test this hypothesis, dynamic light scattering (DLS) was performed on excised vitreous of patients with clinically significant floaters, and compared to macular pucker controls. METHODS: Undiluted, unfixed vitreous was procured during 25-gauge vitrectomy in 14 subjects (age = 59 ± 6.6 years) with clinically significant vitreous floaters, and 6 controls (age = 66.5 ± 8.7 years; P = 0.10) with macular pucker. Total protein concentration was determined by fluorescent Quant-iTTM protein assay kit (Invitrogen/Molecular Probes, Eugene, OR) with bovine serum albumin (0500 ng/ml) as a standard. Fluorescence (excitation at 470 nm and emission at 570 nm) was measured using a Gemini XPS Dual-Scanning Microplate Spectrofluorometer and data analyzed using SoftMax Pro software (Molecular Devices, Sunnyvale, CA). DLS (NS300, Malvern Instruments, Westborough, MA) measurements were performed in each specimen after 10-fold dilution in phosphate buffered saline to optimize concentration in each specimen and determine the mean number of particles, the particle size distributions, and the average particle sizes. RESULTS: Total protein concentration in vitreous specimens trended higher in macular pucker controls (1037 ± 1038 μg/mL) than eyes with vitreous floaters (353.7 ± 141.1 μg/mL; P = 0.08). When normalized to total protein concentration, the number of particles in vitreous from floater eyes was more than 2-fold greater than controls (P < 0.04). Particle size distributions were similarly two-fold greater in vitreous from floater subjects as compared to controls (P < 0.05). The average particle size in vitreous from floater eyes was 315.8 ± 194.6 nm, compared to 147.7 ± 129.3 nm in macular pucker controls (P = 0.039). CONCLUSIONS: Vitreous from eyes with clinically significant floaters contains more particles of larger sizes as compared to controls, likely accounting for the degradation of contrast sensitivity previously found in these patients (Retina 34:1062-8, 2014; IOVS 56:1611–7, 2015; AJO 172:7-12, 2016). DLS could elucidate the underlying molecular abnormalities in patients afflicted with bothersome vitreous floaters and help develop clinical tools to better measure vitreous floaters as well as test the efficacy of various therapies.
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