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微形貌图案化表面促进腱细胞和细胞外胶原的排列。

Microtopographically patterned surfaces promote the alignment of tenocytes and extracellular collagen.

机构信息

Department of Chemical & Biomolecular Engineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.

出版信息

Acta Biomater. 2010 Jul;6(7):2580-9. doi: 10.1016/j.actbio.2009.12.047. Epub 2010 Jan 4.

Abstract

This paper investigates the role of microtopographical features on the cytomorphology, alignment, proliferation and gene expression of tenocytes. We made use of simple microfabrication approaches to create surfaces patterned with topographical features suitable for in vitro studies of tenocytes. These surfaces were composed of glass substrates patterned with polymeric ridges spaced from 50 to 250 microm apart. Our studies demonstrate that the microgrooves differentially impact tenocyte shape, alignment and matrix organization along the direction of grooves. Groove widths significantly influenced cellular alignment, with 50 microm grooved patterns affecting alignment most substantially. Polarized light microscopy demonstrated that mature collagen fibers were denser and more oriented within 50 microm patterns. None of the patterns had a significant effect on the expression of genes linked to proliferation or extracellular matrix synthesis, although time in culture profoundly influenced both gene groups. COMP mRNA expression was moderately increased in tenocytes seeded onto 250 microm grooves, but there was no overall beneficial phenotypic effect of aligned growth. The results of this study indicate that microtopography affects cell density and alignment of tenocytes and leads to the deposition of an aligned collagen matrix, but does not significantly impact matrix gene expression or cell phenotype. These outcomes provide insights into the biology of tendon regeneration, thus providing guidance in the design of clinical procedures for tendon repair.

摘要

本文研究了微形貌特征对腱细胞的细胞形态、排列、增殖和基因表达的作用。我们利用简单的微加工方法,在玻璃基底上制造出具有适合腱细胞体外研究的形貌特征的聚合物脊图案。这些脊的间距为 50 至 250 微米。我们的研究表明,微沟道差异影响腱细胞的形状、沿着沟道方向的排列和基质组织。沟道宽度对细胞排列有显著影响,50 微米的沟道模式影响最大。偏光显微镜显示,成熟的胶原纤维在 50 微米的图案中更加密集和定向。没有一种图案对与增殖或细胞外基质合成相关的基因表达有显著影响,尽管培养时间对这两个基因群都有深远的影响。在接种到 250 微米沟槽中的腱细胞中,COMP mRNA 的表达适度增加,但对齐生长并没有整体有益的表型效应。本研究的结果表明,微形貌影响腱细胞的密度和排列,并导致对齐胶原基质的沉积,但对基质基因表达或细胞表型没有显著影响。这些结果为肌腱再生的生物学提供了新的认识,从而为肌腱修复的临床程序设计提供了指导。

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