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首页> 外文期刊>Journal of Biomechanics >The influence of substrate elastic modulus on retinal pigment epithelial cell phagocytosis
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The influence of substrate elastic modulus on retinal pigment epithelial cell phagocytosis

机译：底物弹性模量对视网膜色素上皮细胞吞噬作用的影响

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摘要

To better understand if a complex process such as phagocytosis is influenced by substrate stiffness, we investigated the influence of substrate elastic modulus on phagocytosis in the retinal pigment epithelial (RPE) cell line ARPE-19. RPE cells lie on Bruch[U+05F3]s membrane, directly under the retina, and phagocytose the shed photoreceptor outer segments. Bruch[U+05F3]s membrane is known to increase in stiffness by an order of magnitude with age and thus, this study has potential relevance in explaining retinal changes in age-related macular degeneration.ARPE-19 cells were plated on laminin-coated polyacrylamide substrates of varying elastic modulus. After 14 days in culture, a solution of latex fluorescent beads suspended in PBS was placed in each well. After an incubation time of 4. h, flow cytometry was performed to determine the number of cells that phagocytosed a bead. The number of ARPE-19 cells that phagocytosed a bead decreased continuously as a function of increasing substrate elastic modulus (. p=0.0135), and this was found to be a linear relationship (slope=-0.03305±0.01104, R2=0.4726 per 10,000 cells).Our results suggest that RPE cells display decreased phagocytosis when grown on firmer substrates, and thus, RPE cells in older eyes, in which Bruch[U+05F3]s membrane is stiffer, may demonstrate decreased phagocytosis. Impaired phagocytosis by RPE cells may contribute to impaired metabolism of photoreceptor outer segments and to development of macular degeneration. Material stiffness may be a critical parameter in the development of neural therapies, including retinal prosthetics and stem cell therapies.
机译：为了更好地了解吞噬作用等复杂过程是否受底物刚度影响，我们研究了视网膜弹性上皮（RPE）细胞系ARPE-19中底物弹性模量对吞噬作用的影响。 RPE细胞直接位于视网膜下方的Bruch [U + 05F3]膜上，并吞噬脱落的感光细胞外部片段。已知Bruch [U + 05F3] s膜的硬度会随着年龄的增长而增加一个数量级，因此，这项研究对于解释与年龄相关的黄斑变性的视网膜变化具有潜在的意义。ARPE-19细胞被铺有层粘连蛋白涂层弹性模量不同的聚丙烯酰胺基材。培养14天后，将悬浮在PBS中的乳胶荧光珠溶液置于每个孔中。在4小时的孵育时间后，进行流式细胞术以确定吞噬珠子的细胞数量。吞噬珠子的ARPE-19细胞数量随着底物弹性模量的增加而连续减少（。p = 0.0135），并且发现这是线性关系（斜率= -0.03305±0.01104，R2 = 0.4726 / 10,000我们的结果表明，RPE细胞在牢固的基质上生长时，吞噬作用会降低，因此，较早出现的Bruch [U + 05F3]膜较硬的眼睛中的RPE细胞可能表现出吞噬作用降低。 RPE细胞吞噬功能受损可能会导致感光器外部片段的代谢受损，并导致黄斑变性的发展。材料刚度可能是神经疗法（包括视网膜假体和干细胞疗法）发展中的关键参数。

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