Surface Elastic Properties of Human Retinal Pigment Epithelium Melanosomes†

Abstract

Atomic force microscope (AFM) imaging and nanoindentation measurements in water were used to probe the mechanical properties of retinal pigment epithelium melanosomes isolated from 14-year-old and 76-year-old donors. Topographic imaging reveals surface roughness similar to previous measurements on dry melanosomes. Force-indentation measurements show different types of responses that were catalogued into four different categories. In these measurements no permanent surface damage of melanosomes was observed as revealed by imaging before and after indentation measurements. The indentation measurements that exhibited nearly elastic responses were used to determine the Young's modulus of melanosomes. The average Young's modulus values are similar for 14-year-old and 76-year-old melanosomes with a somewhat narrower distribution for the 14-year-old sample. These elastic modulus values are considerably higher than the modulus of organelles with cytoplasm (<1 MPa) and approaching values of the modulus of protein crystals (approximately 100 MPa) indicating rather high packing density of biologic material in melanosomes. The width of the Young's modulus distributions is considerable spanning from few megapascals to few tens of megapascals indicating large heterogeneity in the structure. A fraction of the force curves cannot be described by the homogeneous elastic sample model; these force curves are consistent with approximately 10 nm structural heterogeneity in melanosomes. The approach-withdraw hysteresis indicates a significant viscoelasticity, particularly in the samples from the 14-year-old sample. Adhesion of the AFM probe was detected on approximately 3% and approximately 20% of the surface of 14-year-old and 76-year-old samples, respectively. In light of previous studies on these same melanosomes using photoelectron emission microscopy, this adhesion is attributed to the presence of lipofuscin on the surface of the melanosomes. This suggestion indicates that part of the difference in photochemical properties between the old and young melanosomes originates from surface lipofuscin.