Chronic inflammatory responses after implantation of biomaterials can lead to fibrotic encapsulation and failure of implants. The present study was designed to reduce the inflammatory responses to biomaterials by assembling polyelectrolyte multilayers (PEMs) composed of glycosaminoglycans (GAGs) and chitosan (Chi) on glass as model surfaces through layer-by-layer (LBL) technique. Surface plasmon resonance (SPR) and water contact angle (WCA) investigations confirmed the multilayer build-up with alternating deposition of GAGs and Chi layers, while zeta potential measurements showed significant negative charges after multilayer deposition, which further proved the PEM formation. Macrophage adhesion, macrophage spreading morphology, foreign body giant cell (FBGC) formation, as well as β1 integrin expression and interleukin-1β (IL-1β) production were all significantly decreased by GAG-Chi multilayer deposition in comparison to the primary poly (ethylene imine) (PEI) layer. Thereby, the type of GAGs played a pivotal role in inhibiting the inflammatory responses to various extents. Especially heparin (Hep)-Chi multilayers hindered all inflammatory responses to a significantly higher extent in comparison to hyaluronic acid (HA)-Chi and chondroitin sulfate (CS)-Chi multilayer systems. Overall, the present study suggests a great potential of GAG-Chi multilayer coating on implants, particularly the Hep-Chi based systems, to reduce the inflammatory responses.
In general, there is a need for passivation of nanopatterned biomaterial surfaces if cells are intended to interact only with a feature of interest. For this reason self-assembled monolayers (SAM), varying in chain length, are used; they are highly effective in preventing protein adsorption or cell adhesion. In addition, a simple and cost-effective technique to design nanopatterns of various sizes and distances, the so-called nanosphere lithography (NSL), is discussed, which allows the control of cell adhesion and growth depending on the feature dimensions. Combining both techniques results in highly selective nanostructured surfaces, showing that single proteins selectively adsorb on activated nanopatterns. Additionally, adhesion and growth of normal human dermal fibroblasts (NHDF) is strongly affected by the nanostructure dimensions, and it is proven that fibronectin (FN) matrix formation of these cells is influenced, too. Moreover, the FN fibrils are linked to the hexagonally close-packed nanopatterns. As a result, the system presented here can be applied in tissue engineering and implant design due to the fact that the nanopattern dimensions give rise to further modifications and allow the introduction of chemical heterogeneity to guide stem cell differentiation in the future.
Nanostructures play important roles in vivo, where nanoscaled features of extracellular matrix (ECM) components influence cell behavior and resultant tissue formation. This review summarizes some of the recent developments in fostering new concepts and approaches to nanofabrication, such as top-down and bottom-up and combinations of the two. As in vitro investigations demonstrate that man-made nanotopography can be used to control cell reactions to a material surface, its potential application in implant design and tissue engineering becomes increasingly evident. Therefore, we present recent progress in directing cell fate in the field of cell mechanics, which has grown rapidly over the last few years, and in various tissue-engineering applications. The main focus is on the initial responses of cells to nanostructured surfaces and subsequent influences on cellular functions. Specific examples are also given to illustrate the potential nanostructures may have for biomedical applications and regenerative medicine.
The
extracellular matrix (ECM) is a nanostructured environment
that provides chemical, mechanical, and topographical stimuli for
various cellular functions. Here, we introduce the application of
laser interference lithography (LIL) to generate hexagonally arranged
gold nanostructures of three different dimensions on silicon to study
the effect of feature dimensions on human adipose-derived stem cells
(hADSC) in terms of adhesion, growth, and differentiation. Self-assembled
monolayers (SAM) were used to passivate the background silicon surface
with a long-chain polyethylene glycol (PEG), whereas the gold nanostructures
were activated with mercaptoundecanoic acid (MUDA) to direct protein
adsorption and cell adhesive structures to them, only. It was possible
to show that the size and distance of the nanostructures affected
the spreading of hADSC with a decrease of cell size with the increase
of feature dimensions, which corresponded also to the expression of
focal adhesions and presence of the small GTPase RhoA. Effects of
these early events, related to outside-in signal transduction, were
visible by an enhanced cell growth on smaller feature dimensions and
distinct effects on cell differentiation. Because of the precise control
of chemical and topographical cues, the presented system offers great
potential to study effects of material topography on stem cell behavior,
which may pave the way for applications in tailoring surfaces of implants
and tissue engineering scaffolds.
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