Two Dimensional, Spatial Arrangement of Fibronectin Adsorbed to Biomaterials with Different Wettabilities

The effects of concentration, pH and substratum wettability on the two dimensional, spatial arrangement of adsorbed fibronectin are determined. Substrata with different wettability were exposed to a well defined flow of a fibronectin solution (0.1 or 1 mg/ml) in potassium phosphate buffer (pH 7.0 or 4.8) during 120 minutes at a shear rate of 20 s1. Rotary shadowed replicas of the surfaces were examined by transmission electron microscopy. Well defined structures, island-like in character on the low wettability substrata and knotted, reticulated on the high wettability substrata, could be seen in case of adsorption from pH 7.0 and the high concentration solution. Structures became more blurred upon lowering the solution pH and fibronectin concentration. Compared with bovine serum albumin, fibronectin shows smaller island-like structures, but the reticulated structure is thicker than for albumin

Preservation of the Cell-Biomaterial Interface at the Ultrastructural Level

Studying the tissue-biomaterial interface at the ultrastructural level is not without problems. Dissolution of the biomaterial in one of the dehydration or embedding media causes holes and shatter during sectioning or dislodgement of the biomaterial. The fine tuning of the hardness of both biomaterial and embedding medium, as well as the introduction of butyl-2,3- epoxypropylether as an intermediate between the dehydration series and the Epon resin , improving the impregnation , will solve many of the problems mentioned. With this improved technique good results were obtained with materials ranging from teflon, poly(Lactic acid) and polyurethanes to tissue culture polystyrene. No holes, shatter or dislodgement of the biomaterial was observed

Review: Biomaterials for Abdominal Wall Reconstruction

The reconstruction of large abdominal wall defects still is a major surgical problem. Many different techniques have been developed for this purpose, most of which appeared to be unsatisfactory. The lack of sufficient tissue requires the insertion of prosthetic material. Non-absorbable prostheses used to reconstruct abdominal wall defects showed the best results . Polypropylene mesh (PPM) and expanded polytetrafluoroethylene (ePTFE) soft-tissue patch are the most frequently used materials for this purpose . However, PPM induces extensive visceral adhesions and erosion of the skin, whereas ePTFE is insufficiently anchored to the adjacent tissue and therefore both materials are not ideal. As a result of own clinical and experimental studies , we constructed a new prosthesis that combines the favourable properties and avoids the drawbacks of PPM and ePTFE and tested it in an experimental study in the rat. The results are promising and warrant future study to find the ideal non -absorbable prosthesis to reconstruct large abdominal wall defects

Characterization of eukaryotic cell surfaces prior to and after serum protein adsorption by X-ray photoelectron spectroscopy - Fibroblasts, HELA epithelial, and smooth muscle cells

Elemental surface concentration ratios N/C, O/C, and P/C of fibroblasts, HELA epithelial cells, and smooth muscle cells, prior to and after washing in the absence or presence of serum proteins, were determined by X-ray photoelectron spectroscopy. Cell surfaces appeared to adsorb hardly any serum proteins, and the relatively high P/C, as compared to N/C and O/C, elemental surface concentration ratio indicated that the cell surfaces consisted mainly of the phospholipid bilayer, with little or no proteins present. The lack of adsorption of serum proteins to the cell surfaces seems at odds with the common notion that cells require adhesive proteins in order to adhere and spread. However, the adsorption behavior of cellularly produced proteins may be completely different, particularly since they seem to be able to displace adsorbed serum proteins from biomaterials surfaces. Interestingly, only HELA epithelial cells (a tumor cell line) appeared to adsorb a very small amount of proteins.</p

Deformability and collision-induced reorientation enhance cell topotaxis in dense microenvironments

In vivo, cells navigate through complex environments filled with obstacles. Recently, the term 'topotaxis' has been introduced for navigation along topographic cues such as obstacle density gradients. Experimental and mathematical efforts have analyzed topotaxis of single cells in pillared grids with pillar density gradients. A previous model based on active Brownian particles has shown that ABPs perform topotaxis, i.e., drift towards lower pillar densities, due to decreased effective persistence lengths at high pillars densities. The ABP model predicted topotactic drifts of up to 1% of the instantaneous speed, whereas drifts of up to 5% have been observed experimentally. We hypothesized that the discrepancy between the ABP and the experimental observations could be in 1) cell deformability, and 2) more complex cell-pillar interactions. Here, we introduce a more detailed model of topotaxis, based on the Cellular Potts model. To model persistent cells we use the Act model, which mimicks actin-polymerization driven motility, and a hybrid CPM-ABP model. Model parameters were fitted to simulate the experimentally found motion of D. discoideum on a flat surface. For starved D. discoideum, both CPM variants predict topotactic drifts closer to the experimental results than the previous ABP model, due to a larger decrease in persistence length. Furthermore, the Act model outperformed the hybrid model in terms of topotactic efficiency, as it shows a larger reduction in effective persistence time in dense pillar grids. Also pillar adhesion can slow down cells and decrease topotaxis. For slow and less persistent vegetative D. discoideum cells, both CPMs predicted a similar small topotactic drift. We conclude that deformable cell volume results in higher topotactic drift compared to ABPs, and that feedback of cell-pillar collisions on cell persistence increases drift only in highly persistent cells

Influence of glutaraldehyde fixation of cells adherent to solid substrata on their detachment during exposure to shear stress

In order to determine the response of fixed and nonfixed cells adherent to a solid substratum to shear stress, human fibroblasts were allowed to adhere and spread on either hydrophilic glass or hydrophobic Fluoroethylene-propylene (FEP-Teflon) and fixed with glutaraldehyde. Then, the cells were exposed to an incrementally loaded shear stress in a parallel plate flow chamber up to shear stresses of about 500 dynes/cm2, followed by exposure to a liquid-air interface passage. The cellular detachment was compared with the one of nonfixed cells. In case of fixed cells, 50% of the adhering cells detached from FEP-Teflon at a shear stress of 350 dynes/cm2, whereas 50% of the adhering, nonfixed cells detached already at a shear stress of 20 dynes/cm2. No fixed cells detached from glass for shear stresses up to at least 500 dynes/cm2. More than 50% of the nonfixed cells were detached from glass at a shear stress of 350 dynes/cm2. Furthermore, the shape and morphology of fixed cells did not change during the incrementally loaded flow, in contrast to the ones of nonfixed cells, which clearly rounded up prior to detachment.</p

The Influence of a Fibrin-Coating Inside a Biodegradable Poly(DL-Lactide-ε-Caprolactone) Nerve Guide on Peripheral Nerve Regeneration

The aim of this study was to evaluate the effect of a fibrin-coating on the inner surface of a biodegradable poly(DL-lactide-E-caprolactone) nerve guide on the speed and quality of the nerve regeneration. The nerve regeneration and orientation of the nerve fibers, as well as the fibrous tissue formation were evaluated. On the short term, nerve regeneration was slightly faster in the non-coated nerve guide. After longer implantation periods (≥ 4 weeks), nerve regeneration in the fibrin-coated nerve guides was characterized by a severe inflammatory response with large numbers of macrophages and polymorphonuclear cells (PMN\u27s). This study clearly demonstrates that nerve regeneration in a fibrin-coated nerve guide is not faster when compared with a non-coated nerve guide, and that nerve regeneration in the fibrin-coated nerve guide is even worse after longer implantation periods

Peripheral Nerve Regeneration and Functional Nerve Recovery After Reconstruction with a Thin-Walled Biodegradable Poly(DL-lactide-ε-caprolactone) Nerve Guide

The aim of this study was to evaluate functional nerve recovery after reconstruction of a 1 em gap in the sciatic nerve of the rat, with a thin-walled biodegradable poly(DLLA-ε-CL) nerve guide. To evaluate both motor and sensory nerve recovery, walking track analysis and electrostimulation tests were carried out after implantation periods ranging from 3 to 26 weeks post-operatively. The first signs of functional nerve recovery could already be observed after 5 weeks. From the histological analysis, it could be concluded that most of the thin-walled nerve guides had collapsed. Despite collapsing, functional nerve recovery was relatively good after 26 weeks (motor nerve recovery 54% and sensory nerve recovery 100%), probably due to guidance of the regenerating nerve fibers along the outside of the poly(DLLA-ε-CL) nerve guide. This thin-walled nerve guide should, therefore, be used in combination with mechanical support