Evaluation of Polycaprolactone/Gelatin/Chitosan Electrospun Membrane for Peritoneal Adhesion Reduction.

In this study, a novel antiadhesion membrane made of polycaprolactone, gelatin, and chitosan was fabricated using the electrospinning technique. A series of polycaprolactone/gelatin/chitosan (PGC) electrospun membranes with different amounts of chitosan (0%, 0.5%, 1%, and 2% in weight percentage) was synthesized. The physicochemical properties and biocompatibility of the fabricated membranes were examined and compared with the aim to select an effective antiadhesion membrane. Scanning electron microscopy showed that these 4 electrospun membranes had similar fiber diameter and pore area, with no statistical differences between them. Furthermore, the contact angle decreased with increased chitosan content, indicating that chitosan may contribute to increased hydrophilic properties. The in vitro degradation test revealed that the higher chitosan content corresponded to a lower degradation rate in PGC membranes within 7 days. All PGC membranes exhibited similar cell proliferation; however, cell proliferation was lower than tissue culture polystyrene (P < 0.05). To compare antiadhesion ability, the adhesion between the cecum and abdominal wall was created in a rat model. Assessment after implantation of electrospun membranes revealed that PGCs with higher chitosan content (PGC2) had better antiadhesion effects, as evaluated by an adhesion score at day 14 postsurgery. Thus, PGC2 was effective in reducing the formation of tissue adhesion. Therefore, PGC electrospun membrane may provide a potential peritoneal antiadhesion barrier for clinical use.

A Biodegradable Hemostatic Gelatin/Polycaprolactone Composite for Surgical Hemostasis.

Massive bleeding is the leading cause of battlefield-related deaths and the second leading cause of deaths in civilian trauma centers. One of the challenges of managing severe wounds is the need to promote hemostasis as quickly as possible, which can be achieved by using hemostatic dressings. In this study, we fabricated 2 kinds of gelatin/polycaprolactone composites with 2 ratios of gelatin/polycaprolactone, 1:1 and 2:1 (GP11 and GP21, respectively). Scanning electron microscopy revealed that the GP11 composite exhibited rougher and more porous structure than the GP21 composite did. Furthermore, both composites showed similar biocompatibility as that of tissue culture polystyrene. Moreover, both GP composites tended to show a gradual decrease in contact angle to zero within 40 minutes. The in vitro blood plasma coagulation assay revealed that the prothrombin time was significantly longer for the GP composites than it was for the Quikclot composite, whereas the activated partial thromboplastin time of the GP11 composite was significantly shorter than that of the gauze. Furthermore, the GP11 had the largest platelet adsorption of all the composites. The in vivo coagulation test showed an obvious shortening of the bleeding time with the Quikclot and GP21 compared with gauze sample. In conclusion, the GP composites showed superior biocompatibility and hemostasis to the gauze and comparable effects with the Qickclot composite. Therefore, the GP composites have the potential for development as biodegradable surgical hemostatic agents.

Fast isolation and expansion of multipotent cells from adipose tissue based on chitosan-selected primary culture.

Adipose-derived adult stem cells (ASCs) have gained much attention because of their multipotency and easy access. Here we describe a novel chitosan-based selection (CS) system instead of the conventional plastic adherence (PA) to obtain the primary ASCs. The minimal amount of adipose tissue for consistent isolation of ASCs is reduced from 10 mL to 5 mL. The selection is based on the specific interaction between cells and chitosan materials, which separate ASCs by forming spheroids during primary culture. The primary culture period was reduced from 4 days to one day and more ASCs (ten-fold expansion) were achieved in a week. The average duration for obtaining 1 × 10(7) cells takes about seven days from 5 mL of adipose tissue, compared to 14 days using the conventional PA method from 10 mL of adipose tissue. The replicative senescence of CS-ASCs is not evident until the fifteenth passage (vs. eighth for the PA-ASCs). The obtained ASCs (CS-ASCs) have less doubling time for the same passage of cells and show greater stemness than those obtained from the conventional PA method (PA-ASCs). Moreover, CS-ASCs undergo trilineage differentiation more effectively than PA-ASCs. The greater differentiation potential of CS-ASCs may be associated with the enrichment and maintenance of CD271 positive cells by chitosan selection of primary culture.

Characterization and biocompatibility of chitosan nanocomposites.

Chitosan nanocomposites were prepared from chitosan and gold nanoparticles (AuNPs) or silver nanoparticles (AgNPs) of ∼5 nm size. Transmission electron microscopy (TEM) showed the NPs in chitosan did not aggregate until higher concentrations (120-240 ppm). Atomic force microscopy (AFM) demonstrated that the nanocrystalline domains on chitosan surface were more evident upon addition of AuNPs (60 ppm) or AgNPs (120 ppm). Both nanocomposites showed greater elastic modulus, higher glass transition temperature (T(g)) and better cell proliferation than the pristine chitosan. Additionally, chitosan-Ag nanocomposites had antibacterial ability against Staphylococcus aureus. The potential of chitosan-Au nanocomposites as hemostatic wound dressings was evaluated in animal (rat) studies. Chitosan-Au was found to promote the repair of skin wound and hemostasis of severed hepatic portal vein. This study indicated that a small amount of NPs could induce significant changes in the physicochemical properties of chitosan, which may increase its biocompatibility and potential in wound management.

Antibacterial properties of silver nanoparticles in three different sizes and their nanocomposites with a new waterborne polyurethane.

Silver nanoparticles (AgNPs) are strong bactericidal agents but they are also cytotoxic. Embedding them in a polymer matrix may reduce their cytotoxic effect. In the present study, AgNPs in three average sizes were tested for their antibacterial activities and cytotoxicity. Nanocomposites from a new waterborne polyetherurethane (PEU) ionomer and AgNPs were prepared without the use of any crosslinker. It was observed that the antibacterial activity of AgNPs against Escherichia coli started at the effective concentration of 0.1-1 ppm, while that against Staphylococcus aureus started at higher concentrations of 1-10 ppm. Cytotoxicity of AgNPs was observed at the concentration of 10 ppm. AgNPs with smaller average size showed greater antibacterial activity as well as cytotoxicity. The PEU synthesized in this study showed high tensile strength, and the addition of AgNPs at all sizes further increased its thermal stability. The delicate surface features of nanophases, however, were only observed in nanocomposites with either small-or medium-sized AgNPs. PEU-Ag nanocomposites had a strong bacteriostatic effect on the growth of E. coli and S. aureus. The proliferation of endothelial cells on PEU-Ag nanocomposites was enhanced, whereas the platelet adhesion was reduced. The expression of endothelial nitric oxide synthase gene was upregulated on PEU-Ag containing small-sized AgNPs (30 ppm) or medium-sized AgNPs (60 ppm). This effect was not as remarkable in nanocomposites from large-sized AgNPs. Overall, nanocomposites from the PEU and 60 ppm of the medium-sized (5 nm) AgNPs showed the best biocompatibility and antibacterial activity. Addition of smaller or larger AgNPs did not produce as substantial an effect in PEU, especially for the larger AgNPs.

The effect of laser preexposure on seeding endothelial cells to a biomaterial surface.

OBJECTIVE: In this study, the possible effect of low-level laser (LLL) on improving the adhesion of endothelial cells (ECs) to a biomaterial substrate was evaluated. BACKGROUND DATA: Despite the numerous studies regarding the effects of LLL on biologic systems, the influence of LLL on the binding between cells and materials was rarely investigated. MATERIALS AND METHODS: A low-power He-Ne laser apparatus with a continuous wavelength of 632.8 nm (a maximum power output of 50 mW) was used. The average irradiation energy on cells was 1.18 J/cm(2). Cell morphology and the concentrations of nitric oxide and calcium after laser exposure were measured. Biomedical grade poly(carbonate)urethane (PU) was synthesized and used to prepare microporous vascular grafts. ECs exposed to laser were harvested and seeded on the PU grafts. No further exposure was given. RESULTS: LLL could change the morphology and increase the matrix secretion of ECs, and such effects persisted when preexposed cells were harvested and seeded to another substrate. The number of ECs attached on the biomaterial substrate was not affected. Preexposed ECs on the PU graft, however, were, on average, more resistant to flushing (i.e., greater cell retention). CONCLUSION: ECs were pretreated with LLL before being seeded onto the PU biomaterial vascular grafts. The retention of LLL-preexposed ECs on the graft surface was enhanced, but not as significantly as that of ECs preexposed to low-intensity ultrasound.