Scanning electron microscopic, transmission electron microscopic, and confocal laser scanning microscopic observation of fibroblasts cultured on microgrooved surfaces of bulk titanium substrata

Abstract

During this study, microtechnology and plasma etching were used to produce gratings 1.0 (TiD01), 2.0 (TiD02), 5.0 (TiD05), and 10.0 m wide (TiD10) into commercially pure titanium wafers. After incubation of rat dermal fibroblast (RDFs) on these surfaces for 3 days, the cells were observed with scanning electron (SEM), transmission electron (TEM), and confocal laser scanning microscopy (CLSM). Results showed that the RDFs as a whole and their stress fibers oriented strictly parallel to the surface pattern on the TiD01 and TiD02 surfaces. On the TiD05 and TiD10 surfaces, this orientation was not observed. In addition, TEM and CLSM demonstrated that the focal adhesion points (FAP) were located mainly on the surface pattern ridges. TEM revealed that FAP were wrapped occasionally around the edges of the ridges. Only the RDFs on both the TiD05 and TiD10 surfaces protruded into the grooves and possessed FAP on the walls of the grooves. Attachment to the groove floor was observed only on the TiD10 textures. Comparison of these results with earlier observations on microtextured silicone rubber substrata suggests that materialspecific properties do not influence the orientational effect of the surface texture on the observed RDF cellular behavior. The proliferation rate of the RDFs, however, seems to be much higher on titanium than on silicone rubber substrata.Recognizing the possible potential of microtextured implant surfaces, many investigators have studied cellular behavior to various micropatterns produced in glass, polystyrene, silicon, quartz, Epon surface replicas, and silicone rubber surface replicas. 2,3-10 Although many silicone rubber implants do exist, other

Connective‐tissue responses to defined biomaterial surfaces. II. Behavior of rat and mouse fibroblasts cultured on microgrooved substrates

“…Our data suggest that the cells cultured in collagen gel dot colonies on the microgrooved surfaces were well aligned, but lower in density, less spread, and longer and narrower in shape than those grown on the control surfaces. These observations are in general agreement with those of other investigators 3, 15, 24, 32–35, 40–43, 50–63. The in vitro effects of microgrooved substrates are probably based on relationships between microgroove dimensions and the abilities of cells to attach, spread, and migrate.…”

“…However, our observations of stress fibers in 4‐day control cultures suggests that cell source or microgroove depth most likely caused these disparate results. The distribution of focal adhesions in our NIH‐3T3 fibroblasts cultured on microgrooved substrates was in general agreement with these investigators' observations that focal adhesions were limited to areas of cell‐substrate contact 24, 55, 61…”

“…Moreover, the differential channeling of cells grown on microgrooves of varying dimensions agrees with similar observations among RTF cell growth on these substrates. Our finding of impaired stress fiber formation in NIH‐3T3 fibroblasts cultivated on microgrooved substrates is intriguing, and differs from den Braber's observations of stress fibers within dermal fibroblasts after 3 days in culture 55. When evident, stress fibers were aligned in parallel with the orientation of the NIH‐3T3 fibroblasts and microgrooves, an observation that differs from den Braber's, but concurs with others' 24, 41, 42.…”

Osteoblast‐like cell response to bioactive composites—Surface‐topography and composition effects

“…Other studies have reported the alignment of cells with parallel groove formations23 and alterations in cell morphology between smooth and featured areas 24. Furthermore, Singhvi, Stephanopolous, and Wang25 and den Braber et al26 reported differences in cell alignment and attachment for features above and below 50 μm in size, as seen in this study, and Walboomers et al9 observed preferential cell attachment to surface ridges, also as seen in this work. However, more detailed confirmation of this preferential attachment through further techniques such as cross‐sectional imaging is needed.…”

Effect of microtextured surfaces on the performance of percutaneous devices

“…The Brunette studies all used percutaneous parts that were made of nonflexible epoxy material, which was titanium‐coated. It is known that silicone and titanium are different with respect to many surface chemical properties, for instance, it has already been described that different protein adsorption and cell attachment reactions are seen on different materials and that the surface chemistry always interacts with microtopography 3, 20. There is always an interplay between the chemistry and the topography of the materials.…”