Cellulose Nanofibril Formulations Incorporating a Low-Molecular-Weight Alginate Oligosaccharide Modify Bacterial Biofilm Development.

Cellulose nanofibrils (CNFs) from wood pulp are a renewable material possessing advantages for biomedical applications because of their customizable porosity, mechanical strength, translucency, and environmental biodegradability. Here, we investigated the growth of multispecies wound biofilms on CNF formulated as aerogels and films incorporating the low-molecular-weight alginate oligosaccharide OligoG CF-5/20 to evaluate their structural and antimicrobial properties. Overnight microbial cultures were adjusted to 2.8 × 109 colony-forming units (cfu) mL-1 in Mueller Hinton broth and growth rates of Pseudomonas aeruginosa PAO1 and Staphylococcus aureus 1061A monitored for 24 h in CNF dispersions sterilized by γ-irradiation. Two CNF formulations were prepared (20 g m-2) with CNF as air-dried films or freeze-dried aerogels, with or without incorporation of an antimicrobial alginate oligosaccharide (OligoG CF-5/20) as a surface coating or bionanocomposite, respectively. The materials were structurally characterized by scanning electron microscopy (SEM) and laser profilometry (LP). The antimicrobial properties of the formulations were assessed using single- and mixed-species biofilms grown on the materials and analyzed using LIVE/DEAD staining with confocal laser scanning microscopy (CLSM) and COMSTAT software. OligoG-CNF suspensions significantly decreased the growth of both bacterial strains at OligoG concentrations >2.58% (P < 0.05). SEM showed that aerogel-OligoG bionanocomposite formulations had a more open three-dimensional structure, whereas LP showed that film formulations coated with OligoG were significantly smoother than untreated films or films incorporating PEG400 as a plasticizer (P < 0.05). CLSM of biofilms grown on films incorporating OligoG demonstrated altered biofilm architecture, with reduced biomass and decreased cell viability. The OligoG-CNF formulations as aerogels or films both inhibited pyocyanin production (P < 0.05). These novel CNF formulations or bionanocomposites were able to modify bacterial growth, biofilm development, and virulence factor production in vitro. These data support the potential of OligoG and CNF bionanocomposites for use in biomedical applications where prevention of infection or biofilm growth is required.

Ultrapure Wood Nanocellulose-Assessments of Coagulation and Initial Inflammation Potential.

Using a lepirudin-based human whole blood model, we evaluated the initial inflammatory and coagulation responses of dense and porous ultrapure (<50 endotoxin units/grams) cellulose nanofibrils (CNF), of carboxylated grade. The CNF was compared to the wound dressing AquaCel because it is a potential wound-healing material. The porous CNF aerogels induced the strongest coagulation potential measured as prothrombin factor 1.2 (PTF1.2). AquaCel induced the strongest complement response by terminal complement complex (TCC) and surface C3c. All materials activated leukocytes CD11b, while the levels of only 3 of 27 cytokines were significantly changed, limited to (i) an elevation of the monocyte chemoattractant protein-1 (MCP-1/CCL) by the CNF aerogel, (ii) a reduction of eosinophil chemotactic proteins (eotaxin/CCL11) by the CNF aerogel, and (iii) a reduction of platelet-derived growth factor BB (PDGF-BB) by all CNF materials. In conclusion, the CNF materials and AquaCel differently activate coagulation, complement, and cytokines, improving the selection possibilities in various treatment situations of wound healing.

Mechanical characteristics of nanocellulose-PEG bionanocomposite wound dressings in wet conditions.

Wood nanocellulose has been proposed for wound dressing applications partly based on its capability to form translucent films with good liquid absorption capabilities. Such properties are adequate for non-healing and chronic wounds where adequate management of exudates is a requirement. In addition, the translucency will allow to follow the wound development without the necessity to remove the dressing from the wound. Understanding the mechanical properties of nanocellulose films and dressings are also most important for tailoring optimizing wound dressing structures with adequate strength, conformability, porosity and exudate management. Mechanical properties are usually assessed in standard conditions (50% relative humidity, RH), which is not relevant in a wound management situation. In this study we have assessed the mechanical properties of three nanocellulose grades varying in the degree of nanofibrillation. The effect of nanofibrillation and of polyethylene glycol (PEG) addition, on the tensile strength, elongation and elastic modulus were assessed after 24h in water and in phosphate-buffered saline (PBS). The results reveal the behavior of the nanocellulose dressings after wetting and shed light into the development of mechanical properties in environments, which are relevant from a wound management point of view.

The interaction of wood nanocellulose dressings and the wound pathogen P. aeruginosa.

Chronic wounds pose an increasingly significant worldwide economic burden (over £1 billion per annum in the UK alone). With the escalation in global obesity and diabetes, chronic wounds will increasingly be a significant cause of morbidity and mortality. Cellulose nanofibrils (CNF) are highly versatile and can be tailored with specific physical properties to produce an assortment of three-dimensional structures (hydrogels, aerogels or films), for subsequent utilization as wound dressing materials. Growth curves using CNF (diameter <20nm) in suspension demonstrated an interesting dose-dependent inhibition of bacterial growth. In addition, analysis of biofilm formation (Pseudomonas aeruginosa PAO1) on nanocellulose aerogels (20g/m2) revealed significantly less biofilm biomass with decreasing aerogel porosity and surface roughness. Importantly, virulence factor production by P. aeruginosa in the presence of nanocellulose materials, quantified for the first time, was unaffected (p>0.05) over 24h. These data demonstrate the potential of nanocellulose materials in the development of novel dressings that may afford significant clinical potential.