Tailored functionalization of poly(L-lactic acid) substrates at the nanoscale to enhance cell response.

Poly(L-lactic) acid (PLLA) has been widely employed in tissue engineering due to its mechanical properties, biodegradability and biocompatibility. The layer-by-layer (LbL) technique was here proposed as a simple method to impart bioactivity to the surface of PLLA substrates. Aminolysis treatment was applied to introduce amino groups on the surface of PLLA solvent cast films. Then, PLLA films were coated with heparin (HE)/chitosan (CH) multilayer by the LbL technique. Each functionalization step was characterized through physico-chemical and morphological analyses. Aminolysis treatment increased film surface wettability (64.8° ± 2.4° against 74.6° ± 1.3° for untreated PLLA) due to the formation of surface amino groups, which were quantified by acid orange colorimetric assay (0.05 nmol/mm2). After the deposition of 9 layers, the static contact angle varied between values close to 40° C (HE-based layer) and 60 °C (CH-based layer), showing the typical alternate trend of LbL coating. The successful HE/CH deposition was confirmed by ATR-FTIR and X-ray photoelectron spectroscopy (XPS) analyses. Particularly, XPS spectra of coated samples showed the presence of nitrogen (indicative of HE and CH deposition), and sulfur (indicative of HE deposition). The amount of deposited HE was quantified by Taylor's Blue colorimetric method: after the deposition of 19 and 20 layers the HE concentration was around 33 µg/cm2. Finally, in vitro studies performed using HaCaT immortalized human skin keratinocytes, C2C12 immortalized mouse myoblasts and human fibroblasts demonstrated that HE/CH multilayer-coated PLLA is a promising substrate for soft tissue engineering, as cell response may be modulated by changing the surface chemical properties.

Bacterial response to different surface chemistries fabricated by plasma polymerization on electrospun nanofibers.

Control over bacterial attachment and proliferation onto nanofibrous materials constitutes a major challenge for a variety of applications, including filtration membranes, protective clothing, wound dressings, and tissue engineering scaffolds. To develop effective devices, the interactions that occur between bacteria and nanofibers with different morphological and physicochemical properties need to be investigated. This paper explores the influence of fiber surface chemistry on bacterial behavior. Different chemical functionalities were generated on the surface of electrospun polystyrene nanofibers through plasma polymerization of four monomers (acrylic acid, allylamine, 1,7-octadiene, and 1,8-cineole). The interactions of Escherichia coli with the surface modified fibers were investigated through a combination of scanning electron microscopy and confocal laser scanning microscopy. Fiber wettability, surface charge, and chemistry were found to affect the ability of bacterial cells to attach and proliferate throughout the nanofiber meshes. The highest proportion of viable cells attachment occurred on the hydrophilic amine rich coating, followed by the hydrophobic octadiene. The acrylic acid coating rich in carboxyl groups showed a significantly lower attraction of bacterial cells. The 1,8-cineole retained the antibacterial activity of the monomer, resulting with a high proportion of dead isolated cells attached onto the fibers. Results showed that the surface chemistry properties of nanofibrous membranes can be strategically tuned to control bacterial behavior.

Electrospun polystyrene fiber diameter influencing bacterial attachment, proliferation, and growth.

Electrospun materials have been widely investigated in the past few decades as candidates for tissue engineering applications. However, there is little available data on the mechanisms of interaction of bacteria with electrospun wound dressings of different morphology and surface chemistry. This knowledge could allow the development of effective devices against bacterial infections in chronic wounds. In this paper, the interactions of three bacterial species (Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus) with electrospun polystyrene meshes were investigated. Bacterial response to meshes with different fiber diameters was assessed through a combination of scanning electron microscopy (SEM) and confocal microscopy. Experiments included attachment studies in liquid medium but also directly onto agar plates; the latter was aimed at mimicking a chronic wound environment. Fiber diameter was shown to affect the ability of bacteria to proliferate within the fibrous networks, depending on cell size and shape. The highest proliferation rates occurred when fiber diameter was close to the bacterial size. Nanofibers were found to induce conformational changes of rod shaped bacteria, limiting the colonization process and inducing cell death. The data suggest that simply tuning the morphological properties of electrospun fibers may be one strategy used to control biofilm formation within wound dressings.

Electrospun nanofibers as dressings for chronic wound care: advances, challenges, and future prospects.

Chronic non-healing wounds show delayed and incomplete healing processes and in turn expose patients to a high risk of infection. Treatment currently focuses on dressings that prevent microbial infiltration and keep a balanced moisture and gas exchange environment. Antibacterial delivery from dressings has existed for some time, with responsive systems now aiming to trigger release only if infection occurs. Simultaneously, approaches that stimulate cell proliferation in the wound and encourage healing have been developed. Interestingly, few dressings appear capable of simultaneously impairing or treating infection and encouraging cell proliferation/wound healing. Electrospinning is a simple, cost-effective, and reproducible process that can utilize both synthetic and natural polymers to address these specific wound challenges. Electrospun meshes provide high-surface area, micro-porosity, and the ability to load drugs or other biomolecules into the fibers. Electrospun materials have been used as scaffolds for tissue engineering for a number of years, but there is surprisingly little literature on the interactions of fibers with bacteria and co-cultures of cells and bacteria. This Review examines the literature and data available on electrospun wound dressings and the research that is required to develop smart multifunctional wound dressings capable of treating infection and healing chronic wounds.