Liguori Gabriel Romero, Zhou Qihui, Liguori Tácia Tavares Aquinas, Barros Guilherme Garcia, Kühn Philipp Till, Moreira Luiz Felipe Pinho, van Rijn Patrick, Harmsen Martin C
University of Groningen, University Medical Center Groningen, Department of Pathology and Medical Biology, Groningen, Netherlands.
Laboratório de Cirurgia Cardiovascular e Fisiopatologia da Circulação (LIM-11), Instituto do Coração (InCor), Hospital das Clinicas HCFMUSP, Faculdade de Medicina, Universidade de Sao Paulo, Sao Paulo, SP, Brazil.
Stem Cells Int. 2019 May 5;2019:5387850. doi: 10.1155/2019/5387850. eCollection 2019.
Progenitor cells cultured on biomaterials with optimal physical-topographical properties respond with alignment and differentiation. Stromal cells from connective tissue can adversely differentiate to profibrotic myofibroblasts or favorably to smooth muscle cells (SMC). We hypothesized that myogenic differentiation of adipose tissue-derived stromal cells (ASC) depends on gradient directional topographic features.
Polydimethylsiloxane (PDMS) samples with nanometer and micrometer directional topography gradients (wavelength () = 464-10, 990 nm; amplitude () = 49-3, 425 nm) were fabricated. ASC were cultured on patterned PDMS and stimulated with TGF-1 to induce myogenic differentiation. Cellular alignment and adhesion were assessed by immunofluorescence microscopy after 24 h. After seven days, myogenic differentiation was examined by immunofluorescence microscopy, gene expression, and immunoblotting.
Cell alignment occurred on topographies larger than = 1758 nm/ = 630 nm. The number and total area of focal adhesions per cell were reduced on topographies from = 562 nm/ = 96 nm to = 3919 nm/ = 1430 nm. Focal adhesion alignment was increased on topographies larger than = 731 nm/ = 146 nm. Less myogenic differentiation of ASC occurred on topographies smaller than = 784 nm/ = 209 nm.
ASC adherence, alignment, and differentiation are directed by topographical cues. Our evidence highlights a minimal topographic environment required to facilitate the development of aligned and differentiated cell layers from ASC. These data suggest that nanotopography may be a novel tool for inhibiting fibrosis.
在具有最佳物理拓扑特性的生物材料上培养的祖细胞会发生排列和分化。结缔组织的基质细胞可能会不利地分化为促纤维化的肌成纤维细胞，或者有利地分化为平滑肌细胞（SMC）。我们假设脂肪组织来源的基质细胞（ASC）的肌源性分化取决于梯度方向的拓扑特征。
制备具有纳米和微米方向拓扑梯度（波长（λ）= 464 - 10,990 nm；振幅（A）= 49 - 3,425 nm）的聚二甲基硅氧烷（PDMS）样品。将ASC接种在有图案的PDMS上，并用转化生长因子 - 1（TGF - 1）刺激以诱导肌源性分化。24小时后通过免疫荧光显微镜评估细胞排列和粘附。七天后，通过免疫荧光显微镜、基因表达和免疫印迹检查肌源性分化。
在大于λ = 1758 nm / A = 630 nm的拓扑结构上发生细胞排列。每个细胞的粘着斑数量和总面积在λ = 562 nm / A = 96 nm至λ = 3919 nm / A = 1430 nm的拓扑结构上减少。在大于λ = 731 nm / A = 146 nm的拓扑结构上粘着斑排列增加。在小于λ = 784 nm / A = 209 nm的拓扑结构上ASC的肌源性分化较少。
ASC的粘附、排列和分化受拓扑线索指导。我们的证据突出了促进ASC形成排列和分化细胞层所需的最小拓扑环境。这些数据表明纳米拓扑可能是抑制纤维化的一种新工具。
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