Barbucci R, Magnani A, Lamponi S, Pasqui D, Bryan S
CRISMA Department of Chemical and Biosystem Sciences and Technologies, University of Siena, Via Aldo Moro n. 2, Siena 53100, Italy.
Biomaterials. 2003 Mar;24(6):915-26. doi: 10.1016/s0142-9612(02)00425-8.
Surface microfabrication techniques were widely utilised for the spatial control of in vitro cell behaviour. A photo-immobilisation procedure was utilised to create micropatterned surfaces: four different stripe patterns (100, 50, 25 and 10 microm) of hyaluronan (Hyal) and its sulphated derivative (HyalS) on silanised glass substrate were obtained.The morphological analysis showed that the surface topography showed regular stripes of 100, 50, 25 and 10 microm wide and ranging from 300 nm up to 1 microm in thickness. They reproduced the exact photo-mask pattern: glass stripes alternating with polysaccharide ones. On the contrary, Hyal microstructures showed just a topographic pattern as the glass stripes appeared to be covered by a thin layer of the macromolecule by TOF-SIMS. Cell adhesion studies demonstrated that melanocytes adhered and oriented within the first 2h of culture on HyalS microdomains and not on Hyal microstructures where they spread on glass substrate around the patterned area. Double photo-immobilised samples characterised by a 100 microm stripe pattern of Hyal or HyalS on the top of a continuous layer of the two polysaccharides were also created in order to investigate the effect of the topography on cell behaviour. The obtained results demonstrated that melanocytes adhered on HyalS stripes while on the Hyal micropatterned surfaces they spread on silanised glass substrate around the structured area, resulting in the exclusion of the topographic pattern.
表面微加工技术被广泛用于体外细胞行为的空间控制。采用光固定程序制作微图案表面：在硅烷化玻璃基板上获得了四种不同条纹图案（100、50、25和10微米）的透明质酸（Hyal）及其硫酸化衍生物（HyalS）。形态分析表明，表面形貌呈现出宽度为100、50、25和10微米且厚度在300纳米至1微米范围内的规则条纹。它们再现了精确的光掩模图案：玻璃条纹与多糖条纹交替出现。相反，通过飞行时间二次离子质谱（TOF-SIMS）分析，Hyal微观结构仅呈现出一种形貌图案，因为玻璃条纹似乎被一层薄薄的大分子覆盖。细胞黏附研究表明，黑素细胞在培养的最初2小时内黏附并定向于HyalS微区，而不在Hyal微观结构上黏附，它们在图案化区域周围的玻璃基板上扩散。还制备了双光固定样品，其特征是在两种多糖的连续层顶部有100微米宽的Hyal或HyalS条纹图案，以研究形貌对细胞行为的影响。所得结果表明，黑素细胞黏附在HyalS条纹上，而在Hyal微图案化表面上，它们在结构化区域周围的硅烷化玻璃基板上扩散，导致形貌图案被排除在外。
