Block copolymer nanopatterns affect cell spreading: Stem versus cancer bone cells.

Bone healing after a tumor removal can be promoted by biomaterials that enhance the bone regeneration and prevent the tumor relapse. Herein, we obtained several nanopatterns by self-assembly of polystyrene-block-poly-(2-vinylpyridine) (PS-b-P2VP) with different molecular weights and investigated the adhesion and morphology of human bone marrow mesenchymal stem cells (BMMSC) and osteosarcoma cell line (SaOS-2) on these patterns aiming to identify topography and chemistry that promote bone healing. We analyzed > 2000 cells per experimental condition using imaging software and different morphometric descriptors, namely area, perimeter, aspect ratio, circularity, surface/area, and fractal dimension of cellular contour (FDC). The obtained data were used as inputs for principal component analysis, which showed distinct response of BMMSC and SaOS-2 to the surface topography and chemistry. Among the studied substrates, micellar nanopatterns assembled from the copolymer with high molecular weight promote the adhesion and spreading of BMMSC and have an opposite effect on SaOS-2. This nanopattern is thus beneficial for bone regeneration after injury or pathology, e.g. bone fracture or tumor removal.

Antithrombotic and hemocompatible properties of nanostructured coatings assembled from block copolymers.

We describe the antithrombotic properties of nanopatterned coatings created by self-assembly of poly(styrene-block-2-vinylpyridine) (PS-b-P2VP) with different molecular weights. By changing the assembly conditions, we obtained nanopatterns that differ by their morphology (size and shape of the nanopattern) and chemistry. The surface exposition of P2VP block allowed quaternization, i.e. introduction of positive surface charge and following electrostatic deposition of heparin. Proteins (albumin and fibrinogen) adsorption, platelet adhesion and activation, cytocompatibility, and reendothelization capacity of the coatings were assessed and discussed in a function of the nanopattern morphology and chemistry. We found that quaternization results in excellent antithrombotic and hemocompatible properties comparable to heparinization by hampering the fibrinogen adhesion and platelet activation. In the case of quaternization, this effect depends on the size of the polymer blocks, while all heparinized patterns had similar performance showing that heparin surface coverage of 40 % is enough to improve substantially the hemocompatibility.

Bactericidal nanopatterns generated by block copolymer self-assembly.

We describe the bactericidal capacity of nanopatterned surfaces created by self-assembly of block copolymers. Distinct nanotopographies were generated by spin-coating with polystyrene-block-poly(2-vinylpyridine) (PS-b-P2VP) followed by solvent vapor annealing. We demonstrate that the bactericidal efficiency of the developed coatings depends on the morphology and the chemistry of the surface: cylindrical nanotopographies presenting both blocks at the surface have stronger bactericidal effect on Escherichia coli than micellar patterns with only PS exposed at the surface. The identified mechanism of bacterial death is a mechanical stress exerted by the nanostructures on the cell-wall. Moreover, the developed nanopatterns are not cytotoxic, which makes them an excellent option for coating of implantable materials and devices. The proposed approach represents an efficient tool in the fight against bacteria, which acts via compromising the bacterial wall integrity. STATEMENT OF SIGNIFICANCE: Bacterial infections represent an important risk during biomaterial implantation in surgeries due to the increase of antibiotic resistance. Bactericidal surfaces are a promising solution to avoid the use of antibiotics, but most of those systems do not allow mammalian cell survival. Nanopatterned silicon surfaces have demonstrated to be simultaneously bactericidal and allow mammalian cell culture but are made by physical methods (e.g. plasma etching) applicable to few materials and small surfaces. In this article we show that block copolymer self-assembly can be used to develop surfaces that kill bacteria (E. coli) but do not harm mammalian cells. Block copolymer self-assembly has the advantage of being applicable to many different types of substrates and large surface areas.

Extracellular matrix-inspired assembly of glycosaminoglycan-collagen fibers.

We report on the fabrication of fibers exclusively from the extracellular matrix components by interfacial complexation without using any crosslinking agent. The obtained fibers may have different biomedical applications as they are flexible, and cells adhere and remodel them. Moreover, their stability and thickness can be finely tuned by the sulfation degree of the used glycosaminoglycans.

A new route to produce starch-based fiber mesh scaffolds by wet spinning and subsequent surface modification as a way to improve cell attachment and proliferation.

This study proposes a new route for producing fiber mesh scaffolds from a starch-polycaprolactone (SPCL) blend. It was demonstrated that the scaffolds with 77% porosity could be obtained by a simple wet-spinning technique based on solution/precipitation of a polymeric blend. To enhance the cell attachment and proliferation, Ar plasma treatment was applied to the scaffolds. It was observed that the surface morphology and chemical composition were significantly changed because of the etching and functionalization of the fiber surfaces. XPS analyses showed an increase of the oxygen content of the fiber surfaces after plasma treatment (untreated scaffolds O/C:0.32 and plasma-treated scaffolds O/C:0.41). Both untreated and treated scaffolds were examined using a SaOs-2 human osteoblast-like cell line during 2 weeks of culture. The cell seeded on wet-spun SPCL fiber mesh scaffolds showed high viability and alkaline phosphatase enzyme activity, with those values being even higher for the cells seeded on the plasma-treated scaffolds.

Surface modification of starch based blends using potassium permanganate-nitric acid system and its effect on the adhesion and proliferation of osteoblast-like cells.

The surface modification of three starch based polymeric biomaterials, using a KMnO4/HNO3 oxidizing system, and the effect of that modification on the osteoblastic cell adhesion has been investigated. The rationale of this work is as follows--starch based polymers have been proposed for use as tissue engineering scaffolds in several publications. It is known that in biodegradable systems it is quite difficult to have both cell adhesion and proliferation. Starch based polymers have shown to perform better than poly-lactic acid based materials but there is still room for improvement. This particular work is aimed at enhancing cell adhesion and proliferation on the surface of several starch based polymer blends that are being proposed as tissue engineering scaffolds. The surface of the polymeric biomaterials was chemically modified using a KMnO4/HNO3 system. This treatment resulted in more hydrophilic surfaces, which was confirmed by contact angle measurements. The effect of the treatment on the bioactivity of the surface modified biomaterials was also studied. The bioactivity tests, performed in simulated body fluid after biomimetic coating, showed that a dense film of calcium phosphate was formed after 30 days. Finally, human osteoblast-like cells were cultured on unmodified (control) and modified materials in order to observe the effect of the presence of higher numbers of polar groups on the adhesion and proliferation of those cells. Two of the modified polymers presented changes in the adhesion behavior and a significant increase in the proliferation rate kinetics when compared to the unmodified controls.