Quantitative analysis of cell proliferation and orientation on substrata with uniform parallel surface micro-grooves

Abstract

In order to quantify the effect of the substrata surface topography on cellular behaviour, planar and micro-textured silicon substrata were produced and made suitable for cell culture by radio frequency glow discharge treatment. These substrata possessed parallel surface grooves with a groove and ridge width of 2.0 (SÍID02), 5.0 (SÍID05) and 10/¿m (SÍID10). Groove depth was approximately 0 .5 /m Rat dermal fibroblasts (RDFs) were cultured on these substrata and a tissue culture polystyrene control surface for 1, 2, 3, 5 and 7 days. After incubation the cell proliferation was quantified with a Coulter Counter, and RDF size, shape and orientation with digital image analysis. Cell counts proved that neither the presence of the surface grooves nor the dimension of these grooves had an effect on the cell proliferation. However, RDFs on SIID02, and to a lesser extent on SÜD05 substrata, were elongated and aligned parallel to the surface grooves. Orientation of the RDFs on SÍID10 substrata proved to be almost comparable to the SIID00 substrata. Finally, it was observed that the cells on the micro-textured substrata were capable of spanning the surface grooves. © 1996 Elsevier Science Limited Correspondence to E.T. den Braber.Most of the studies are focussed on the role of contact guidance in fundamental organogenesis phenomena like surface implanted biomaterials has only been recognized for last few years. Brunette et a l"'l0, for example, the micro-e implant surfaces to prevent epithelial downgrowth around skin-penetrating devices. Campbell and von Rocum11 described the use of surface micro-patterns as a tool to implantreduce the inflammatory response at tissue interface. Although these studies have provided important information, the fundamental mechanism of, and optimal parameters for, cell control by guidance are still unknown. In addition, the reported results are often based on subjective, qualitative observations. To surpass this lack of knowledge it is evident that a systematic study of the influence of surface topography on the cellular behaviour is required. Therefore, the objective of our studies is to approach this guidance principle in a more orderly way.In our first study12 we reported on the surface treatment on the wettability of surfaces and on 1093

Effect of surface roughness on initial responses of osteoblast-like cells on two types of zirconia

“…The surface roughness of materials can profoundly affect cell attachment and spreading, and a roughened surface is generally thought to be preferable for cell attachment 40,43) . Furthermore, initial cell adhesion on the material surface occurs through mechanical interlocking 44) . Therefore, an appropriate surface roughness can produce beneficial mechanical interlocking at the initial adhesion stage and aid in future cell adhesion.…”

Maintenance of Human Hepatocyte FunctionIn Vitroby Liver-Derived Extracellular Matrix Gels

“…39 Well-known biophysical properties such as three-dimensional (3D) surface topography play important roles in cell phenotype and behavior. [40][41][42][43][44] The 3D surface topography of an ECM includes variation in pore diameter as well as variation in the distances between the pores, 45 surface texture, 43 hydrophobicity and=or hydrophilicity, and the presence or absence of a basement membrane. 24 Obviously, the 3D surface topography of the gel form of PLECM differs greatly from that of the native liver ECM.…”

Orientation of ECM protein deposition, fibroblast cytoskeleton, and attachment complex components on silicone microgrooved surfaces

“…For the production of the substrata for in vitro purposes, the smooth (SilD00) and microgrooved silicon molds were covered with polydimethyl-siloxane (silicone elastomer MDX 4-4210, Dow Corning). After polymerization, the silicone rubber negative surface replicas were removed, cut into appropriate circular shapes and sizes (175 mm 2 ), mounted on glass microscopic slides (76 × 26 mm; Knittel, Germany), cleaned ultrasonically with Liquinox and by Soxhlet rinsing, as described before, [4][5][6] and prepared for cell culture purposes by radio frequency glow discharge (RFGD) treatment (PDC-3XG, Harrick; Argon, 0.15 Torr, 5 min).…”