Faculty Bibliography

Surface microgeometry strongly influences the shapes, orientations, and growth characteristics of cultured cells, but in-depth, quantitative studies of these effects are lacking. We investigated several contact guidance effects in cells within "dot" colonies of primary fibroblasts and in cultures of a transformed fibroblast cell line, employing titanium-coated, microgrooved polystyrene surfaces that we designed and produced. The aspect ratios, orientations, densities, and attachment areas of rat tendon fibroblasts (RTF) colony cells, in most cases, varied (p < 0.01) by microgroove dimension. We observed profoundly altered cell morphologies, reduced attachment areas, and reduced cell densities within colonies grown on microgrooved substrates, compared with cells of colonies grown on flat, control surfaces. 3T3 fibroblasts cultured on microgrooved surfaces demonstrated similarly altered morphologies. Fluorescence microscopy revealed that microgrooves alter the distribution and assembly of cytoskeletal and attachment proteins within these cells. These findings are consistent with previous results, and taken together with the results of our in vivo and cell colony growth studies, enable us to propose a unified hypothesis of how microgrooves induce contact guidance.

Recent studies have shown that it is now possible to construct tissue-engineered bone repair scaffolds with tight pore size distributions and controlled geometries using 3-D Printing techniques (3DP). This study evaluated two hydroxyapatite (HA) 8-mm diameter discs with controlled architectures in a rabbit trephine defect at 8 and 16 weeks using a 2 x 2 factorial design. Input parameters were time and scaffold void volume at two levels. Three output variables were extracted from MicroCT data: bone volume ingrowth with respect to total region of interest, bone volume ingrowth with respect to available ingrowth volume, and soft tissue volume. The experiment measured two groups--Group 1: 500-microm x 500-microm channels parallel to the scaffold's long axis and penetrating up 3-mm from the bottom. Group 2: 800-microm x 800-microm struts spaced 500 microm apart set perpendicularly to each other in each printed layer. Rendered 3-dimensional MicroCT scans and undecalcified histological slides of implants revealed good integration with the surrounding tissue, and a sizeable amount of bone ingrowth into the device. Factorial analysis revealed that the effects of time were the greatest determinant of soft tissue ingrowth, while time and its interaction with void volume were the greatest determinants of bone volume ingrowth with respect to both total and available volume.

Surface microgeometry plays a role in tissue-implant surface interactions, but our understanding of its effects is incomplete. Substrate microgrooves strongly influence cells in vitro, as evidenced by contact guidance and cell alignment. We studied "dot" colonies of primary fibroblasts and bone marrow cells that were grown on titanium-coated, microgrooved polystyrene surfaces that we designed and produced. Rat tendon fibroblast and rat bone marrow colony growth and migration varied (p < 0.01) by microgroove dimension and slightly by cell type. We observed profoundly altered morphologies, reduced growth rates, and directional growth in colonies grown on microgrooved substrates, when compared with colonies grown on flat, control surfaces (p < 0.01). The cells in our colonies grown on microgrooved surfaces were well aligned and elongated in the direction parallel to the grooves and colonies. Our "dot" colony is an easily reproduced, easily measured and artificial explant model of tissue-implant interactions that better approximates in vivo implant responses than culturing isolated cells on biomaterials. Our results correlate well with in vivo studies of titanium dioxide-coated polystyrene, titanium, and titanium alloy implants with controlled microgeometries. Microgrooves and other surface features appear to directionally or spatially organize cells and matrix molecules in ways that contribute to improved stabilization and osseointegration of implants.

The in vivo bone response of 3D periodic hydroxyapatite (HA) scaffolds is investigated. Two groups of HA scaffolds (11 mm diameter x 3.5 mm thick) are fabricated by direct-write assembly of a concentrated HA ink. The scaffolds consist of cylindrical rods periodically arranged into four quadrants with varying separation distances between rods. In the first group, HA rods (250 microm in diameter) are patterned to create pore channels, whose areal dimensions are 250 x 250 microm(2) in quadrant 1, 250 x 500 microm(2) in quadrants 2 and 4, and 500 x 500 microm(2) in quadrant 3. In the second group, HA rods (400 microm in diameter) are patterned to create pore channels, whose areal dimensions of 500 x 500 microm(2) in quadrant 1, 500 x 750 microm(2) in quadrants 2 and 4, and 750 x 750 microm(2) in quadrant 3. Each group of scaffolds is partially densified by sintering at 1200 degrees C prior to being implanted bilaterally in trephine defects of skeletally mature New Zealand White rabbits. Their tissue response is evaluated at 8 and 16 weeks using micro-computed tomography, histology, and scanning electron microscopy. New trabecular bone is conducted rapidly and efficiently across substantial distances within these patterned 3D HA scaffolds. Our observations suggest that HA rods are first coated with a layer of new bone followed by subsequent scaffold infilling via outward and inward radial growth of the coated regions. Direct-write assembly of 3D periodic scaffolds composed of micro-porous HA rods arrayed to produce macro-pores that are size-matched to trabecular bone may represent an optimal strategy for bone repair and replacement structures.