State Key Laboratory of New Ceramics and Fine Processing, Department of Materials Science and Engineering, Tsinghua University, Beijing 100084, China.
J Tissue Eng Regen Med. 2013 Feb;7(2):112-7. doi: 10.1002/term.498. Epub 2011 Dec 12.
This study investigated how human adipose stem cells (hASCs) could be influenced by surface chemistry. Self-assembled monolayers of alkanethiolates on gold were introduced as a surface chemistry model to provide a range of functional groups such as OH, COOH, NH₂, Phenyl, SH, Br, and CH₃ on surfaces. Initially, morphological changes of hASCs in response to different surface chemistries were observed with focal adhesion. Cell growth behaviour evaluated by Cell Counting Kit-8 (CCK8) assay (Dojindo Molecular Technologies Inc., Shanghai, China) and cytoskeletal F-actin Biochem Kit™ (Denver, CO, USA) staining revealed a descending order of growth rate on the following surfaces: NH₂ > SH > COOH > Phenyl > Br > OH > CH₃. The mRNA expressions of lineage specific markers including alkaline phosphatase (ALP), osteocalcin (OCN), type II collagen, aggrecan, peroxisome proliferator-activated receptor gamma (PPARγ), and fatty acid binding protein-2 (aP2), were determined using real-time reversed transcriptase-polymerase chain reaction (RT-PCR). Results revealed that NH₂ favoured hASC differentiation toward osteogenic, while phenyl and SH promoted chondrogenic differentiation of hASCs with a high level up-regulation of type II collagen and aggrecan. hASCs on Br increased in PPARγ and aP2 expression, indicating adipogenic differentiation. These results highlight the vital role of surface chemistry on the modulation of hASC differentiation and suggests chemical methods for designing biomaterials for stem cell-based tissue regeneration.
本研究探讨了人脂肪干细胞（hASCs）如何受到表面化学的影响。金的自组装单分子层烷硫醇用作表面化学模型，在表面上提供了一系列官能团，例如 OH、COOH、NH₂、Phenyl、SH、Br 和 CH₃。最初，通过焦点黏附观察到 hASCs 对不同表面化学的形态变化。通过细胞计数试剂盒-8（CCK8）测定（Dojindo Molecular Technologies Inc.，上海，中国）和细胞骨架 F-肌动蛋白生化试剂盒™（丹佛，CO，美国）染色评估细胞生长行为显示以下表面上的生长速率呈下降趋势：NH₂>SH>COOH>Phenyl>Br>OH>CH₃。使用实时逆转录-聚合酶链反应（RT-PCR）测定谱系特异性标志物包括碱性磷酸酶（ALP）、骨钙素（OCN）、II 型胶原、聚集蛋白聚糖、过氧化物酶体增殖物激活受体 γ（PPARγ）和脂肪酸结合蛋白-2（aP2）的 mRNA 表达。结果表明，NH₂有利于 hASC 向成骨方向分化，而苯基和 SH 促进 hASCs 向软骨分化，II 型胶原和聚集蛋白聚糖的高表达上调。Br 上的 hASCs 增加了 PPARγ和 aP2 的表达，表明其向脂肪细胞分化。这些结果突出了表面化学对 hASC 分化的调节作用，并为基于干细胞的组织再生的生物材料设计提供了化学方法。
