BioMEMS Laboratory, Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic, 9500 Euclid Avenue, Cleveland, OH 44195, USA.
Acta Biomater. 2010 Jan;6(1):160-9. doi: 10.1016/j.actbio.2009.06.016. Epub 2009 Jun 16.
The influence of surface microtexture on osteogenesis was investigated in vitro by examining the proliferation and differentiation characteristics of a class of adult stem cells and their progeny, collectively known as connective tissue progenitor cells (CTPs). Human bone marrow-derived CTPs were cultured for up to 60 days on smooth polydimethylsiloxane (PDMS) surfaces and on PDMS with post microtextures that were 10 microm in diameter and 6 microm in height, with 10 microm separation. DNA quantification revealed that the numbers of CTPs initially attached to both substrates were similar. However, cells on microtextured PDMS transitioned from lag phase after 4 days of culture, in contrast to 6 days for cells on smooth surfaces. By day 9 cells on the smooth surfaces exhibited arbitrary flattened shapes and migrated without any preferred orientation. In contrast, cells on the microtextured PDMS grew along the array of posts in an orthogonal manner. By days 30 and 60 cells grew and covered all surfaces with extracellular matrix. Western blot analysis revealed that the expression of integrin alpha5 was greater on the microtextured PDMS compared with smooth surfaces. Real time reverse transcription-polymerase chain reaction revealed that gene expression of alkaline phosphatase had decreased by days 30 and 60, compared with that on day 9, for both substrates. Gene expression of collagen I and osteocalcin was consistently greater on post microtextures relative to smooth surfaces at all time points.
研究人员通过观察一类成体干细胞及其后代(统称为结缔组织祖细胞)的增殖和分化特性,研究了表面微观结构对成骨的影响。将人骨髓来源的结缔组织祖细胞在光滑的聚二甲基硅氧烷(PDMS)表面和具有 10 微米直径和 6 微米高度、间隔 10 微米的后微观结构的 PDMS 上培养长达 60 天。DNA 定量分析显示,最初附着在两种基质上的 CTP 数量相似。然而,在培养 4 天后,微纹理 PDMS 上的细胞从滞后期转变,而光滑表面上的细胞则需要 6 天。到第 9 天,光滑表面上的细胞呈现出任意的扁平形状,并在没有任何优先取向的情况下迁移。相比之下,微纹理 PDMS 上的细胞以正交方式沿柱阵列生长。在第 30 天和第 60 天,细胞生长并覆盖了所有表面的细胞外基质。Western blot 分析显示,与光滑表面相比,微纹理 PDMS 上整合素 alpha5 的表达更高。实时逆转录-聚合酶链反应显示,与第 9 天相比,两种基质上的碱性磷酸酶基因表达在第 30 天和第 60 天都有所下降。在所有时间点,微纹理柱上的 I 型胶原和骨钙素基因表达都明显高于光滑表面。