Yuyu Sun

Silver sulfadiazine–immobilized celluloses as biocompatible polymeric biocides

Sulfadiazine was immobilized onto cotton cellulose using ethylene glycol diglycidyl ether as a binder. Upon treatment with diluted silver nitrate aqueous solution, the sulfadiazine moieties in the immobilized celluloses were transformed into silver–sulfadiazine coordination complexes. The resulting silver sulfadiazine–immobilized celluloses provided a 6-log reduction of 108 CFU mL−1 of Staphylococcus aureus (Gram-positive bacteria), Escherichia coli (Gram-negative bacteria), methicillin-resistant Staphylococcus aureus (drug-resistant bacteria), vancomycin-resistant Enterococcus faecium (drug-resistant bacteria), and Candida albicans (fungi) in 30–60 minutes, and a 5-log reduction of 107 PFU mL−1 of MS2 virus in 120 minutes. The antibacterial, antifungal, and antiviral activities were both durable and rechargeable. Additionally, trypan blue assay suggested that the new silver sulfadiazine–immobilized celluloses sustained excellent mammal cell viability, pointing to great potentials of the new materials for a broad range of health care–related applications.

Novel Anti-infective Activities of Chitosan Immobilized Titanium Surface with Enhanced Osteogenic Properties

We have covalently immobilized chitosan onto a titanium (Ti) surface to manage implant-related infection and poor osseointegration, two of the major complications of orthopedic implants. The Ti surface was first treated with sulfuric acid (SA) and then covalently grafted with chitosan. Surface roughness, contact angle and surface zeta potential of the samples were markedly increased by the sulfuric acid treatment and the subsequent chitosan immobilization. The chitosan-immobilized Ti (SA-CS-Ti) showed two novel antimicrobial roles: it (a) prevented the invasion and internalization of bacteria into the osteoblast-like cells, and (b) significantly increased the susceptibility of adherent bacteria to antibiotics. In addition, the sulfuric acid-treated Ti (SA-Ti) and SA-CS-Ti led to significantly increased (P<0.05) osteoblast-like cell attachment, enhanced cell proliferation, and better osteogenic differentiation and mineralization of osteoblast-like cells.

Acyclic N-halamine-immobilized polyurethane: Preparation and antimicrobial and biofilm-controlling functions

Hydroxyl groups were introduced onto polyurethane surfaces through 1,6-hexamethylene diisocyanate activation, followed by diethanolamine hydroxylation. Polymethacrylamide was covalently attached to the hydroxylated polyurethane through surface grafting polymerization of methacrylamide using cerium (IV) ammonium nitrate as an initiator. After bleach treatment, the amide groups of the covalently bound polymethacrylamide chains were transformed into N-halamines. The new N-halamine-immobilized polyurethane provided a total sacrifice of 107−108 colony forming units per milliliter of Staphylococcus aureus (Gram-positive bacteria), Escherichia coli (Gram-negative bacteria), and Candida albicans (fungi) within 10 min and successfully prevented bacterial and fungal biofilm formation. The antimicrobial and biofilm-controlling effects were both durable and rechargeable, pointing to great potentials of the new acyclic N-halamine-immobilized polyurethane for a broad range of related applications.

Controlling fungal biofilms with functional drug delivery denture biomaterials.

Candida-associated denture stomatitis (CADS), caused by colonization and biofilm-formation of Candida species on denture surfaces, is a significant clinical concern. We show here that modification of conventional denture materials with functional groups can significantly increase drug binding capacity and control drug release rate of the resulting denture materials for potentially managing CADS. In our approach, poly(methyl methacrylate) (PMMA)-based denture resins were surface grafted with three kinds of polymers, poly(1-vinyl-2-pyrrolidinone) (PNVP), poly(methacrylic acid) (PMAA), and poly(2-hydroxyethyl methacrylate) (PHEMA), through plasma-initiated grafting polymerization. With a grafting yield as low as 2 wt%, the three classes of new functionalized denture materials showed significantly higher drug binding capacities toward miconazole, a widely used antifungal drug, than the original PMMA denture resin control, leading to sustained drug release and potent biofilm-controlling effects against Candida. Among the three classes of functionalized denture materials, PNVP-grafted resin provided the highest miconazole binding capability and the most powerful antifungal and biofilm-controlling activities. Drug binding mechanisms were studied. These results demonstrated the importance of specific interactions between drug molecules and functional groups on biomaterials, shedding lights on future design of CADS-managing denture materials and other related devices for controlled drug delivery.

Bacteria and osteoblast adhesion to chitosan immobilized titanium surface: A race for the surface.

In order to evaluate the anti-infective efficacy of the titanium implant materials, two co-culture systems, a low-bacteria/osteoblast (L-B) and a high-bacteria/osteoblast system (H-B), were established. Untreated (UN-Ti), sulfuric acid-treated (SA-Ti), and chitosan immobilized titanium (SA-CS-Ti) materials were developed and evaluated. Bacteria and osteoblast behaviors, including initial attachment (evaluated at 30 mins), adhesion (evaluated at 4 h), and osteoblast spreading on each material surface were evaluated using quantification assays, scanning electron microscopy (SEM), and confocal microscopy. Quantification analysis at 30 mins showed significantly higher number of osteoblast present on SA-CS-Ti in both L-B (10,083 ± 2626) and H-B (23,592 ± 2233) than those on the UN-Ti (p<0.05). SEM observation and confocal microscopy results showed more surface area was occupied by adhered osteoblasts on SA-CS-Ti than UN-Ti and SA-Ti in both co-culture systems at 30 mins. At all time points, SA-CS-Ti had the lowest level of bacterial adhesion compared to UN-Ti and SA-Ti in both co-culture systems. A significantly (p<0.05) lower number of bacteria were recovered from SA-CS-Ti (2233 ± 681) in the H-B system compared to UN-Ti (5367 ± 1662) and SA-Ti (4533 ± 680) at 4h. Quantitative and qualitative co-culture results show the great potential of chitosan immobilization onto implant materials to prevent implant-associated infections.

N‐trimethylchitosan/Alginate Layer‐by‐Layer Self Assembly Coatings Act as “Fungal Repellents” to Prevent Biofilm Formation on Healthcare Materials

Fungal biofilm formation on healthcare materials is a significant clinical concern, often leading to medical-device-related infections, which are difficult to treat. A novel fungal repellent strategy is developed to control fungal biofilm formation. Methylacrylic acid (MAA) is grated onto poly methyl methacrylate (PMMA)-based biomaterials via plasma-initiated grafting polymerization. A cationic polymer, trimethylchitosan (TMC), is synthesized by reacting chitosan with methyl iodide. Sodium alginate (SA) is used as an anionic polymer. TMC/SA multilayers are coated onto the MAA-grafted PMMA via layer-by-layer self-assembly. The TMC/SA multilayer coatings significantly reduce fungal initial adhesion, and effectively prevent fungal biofilm formation. It is concluded that the anti-adhesive property of the surface is due to its hydrophilicity, and that the biofilm-inhibiting action is attributed to the antifungal activity of TMC as well as the chelating function of TMC and SA, which may have acted as fungal repellents. Phosphate buffered saline (PBS)-immersion tests show that the biofilm-modulating effect of the multilayer coatings is stable for more than 4 weeks. Furthermore, the presence of TMC/SA multilayer coatings improves the biocompatibility of the original PMMA, offering a simple, yet effective, strategy for controlling fungal biofilm formation.

Cytocompatible antibacterial fibrous membranes based on poly(3-hydroxybutyrate-co-4-hydroxybutyrate) and quaternarized N-halamine polymer

A novel polymeric N-halamine-containing quaternary ammonium salt (PHQS) was synthesized and used to make antibacterial electrospun fibrous membranes by blending with biodegradable poly(3-hydroxybutyrate-co-4-hydroxybutyrate) (P(3HB-4HB)). The chemical structures of 3-(2′-chloroethyl)-5,5-dimethylhydantoin (CEDMH), poly(2-(dimethylamino)ethyl methacrylate) (PDMAEMA) and PHQS were characterized with FT-IR, 1H NMR, 13C NMR and GPC. The obtained electrospun fibrous membranes were chlorinated with chlorine bleach and characterized by scanning electron microscopy (SEM) and thermogravimetry (TG). The new fibrous membranes provided potent antimicrobial activities against Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli O157:H7. In addition, the treated electrospun fibrous membranes showed excellent stability and durability in UVA light irradiation and storage tests. The results of rat skin fibroblast cytotoxicity studies indicated that the antimicrobial membranes are biocompatible. From this research, polymeric quaternarized N-halamine antimicrobial fibrous membranes based on P(3HB-4HB) may have potential use as eco-friendly materials in food packaging and biomedicine.

Quaternized chitosans bind onto preexisting biofilms and eradicate pre-attached microorganisms.

Quaternized chitosans, N,N,N-trimethylchitosans (TMC) with different degree of quaternization were synthesized by reacting methyl iodide with chitosan. The reaction was confirmed by FT-IR and 1H-NMR characterization. Antimicrobial assay showed that the prepared TMC had potent biocidal effects against planktonic Gram-positive bacteria Staphylococcus epidermidis, Gram-negative bacteria Escherichia coli, and yeast Candida albicans. Bacterial and fungal biofilms were formed on poly(methyl methacrylate) (PMMA) films and then treated with TMC aqueous solutions. Zeta potential measurement suggested that TMC bonded onto the preexisting biofilms. Biofilm-binding kinetics was evaluated in UV studies using phenyl group-labeled TMC as model compounds, which revealed that quaternized chitosans bonded onto the preexisting biofilms rapidly. Colony-forming unit (CFU) determination and SEM, confocal laser scanning microscopy (CLSM) and fluorescence microscopy studies demonstrated that the bonded TMC had powerful biocidal activities to eradicate the pre-attached bacterial and fungal cells in the preexisting biofilms. The biocompatibility of the TMC samples with rat skin fibroblast cells was evaluated in the MTT assay.

A comparison of 2 laboratory methods to test dental unit waterline water quality.

The performance of 2 American Public Health Association standard laboratory methods, the R2A spread plate and the SimPlate(TM) for heterotrophic plate count, for quantifying heterotrophic microorganisms in dental waterline samples was evaluated. Microbial counts were underestimated on SimPlate(TM) compared with R2A, and the results indicated a poor correlation between the 2 methods.

Cellulose-ethylenediaminetetraacetic acid conjugates protect mammalian cells from bacterial cells.

Cellulose-ethylenediaminetetraacetic acid (EDTA) conjugates were synthesized by the esterification of cellulose with ethylenediaminetetraacetic dianhydride (EDTAD). The new materials provided potent antimicrobial activities against Staphylococcus aureus (S. aureus, Gram-positive bacteria) and Pseudomonas aeruginosa (P. aeruginosa, Gram-negative bacteria), and inhibited the formation of bacterial biofilms. The biocompatibility of the new cellulose-EDTA conjugates was evaluated with mouse skin fibroblasts for up to 14 days. SEM observation and DNA content analysis suggested that the new materials sustained the viability of fibroblast cells. Moreover, in mouse skin fibroblast-bacteria co-culture systems, the new cellulose-EDTA conjugates prevented bacterial biofilm formation and protected the mammalian cells from the bacterial cells for at least one day.