Meilin Yang

Electrospun elastin-like polypeptide enriched polyurethanes and their interactions with vascular smooth muscle cells

In vascular tissue, elastin is an essential extracellular matrix protein that plays an important biomechanical and biological signalling role. Native elastin is insoluble and is difficult to extract from tissues, which results in its relatively rare use for the fabrication of vascular tissue engineering scaffolds. Recombinant elastin-like polypeptide-4 (ELP4), which mimics the structure and function of native tropoelastin, represents a practical alternative to the native elastic fibre for vascular applications. In this study, electrospinning was utilized to fabricate fibrous scaffolds which were subsequently surface modified with ELP4 and used as substrates for smooth muscle cell culture. ELP4 surface modified materials demonstrated enhanced smooth muscle cell (SMC) adhesion and maintenance of cell numbers over a 1-week period relative to controls. SMCs seeded on the ELP4 surface modified materials were also shown to exhibit the cell morphology and biological markers of a contractile phenotype including a spindle-like morphology, actin filament organization and smooth muscle myosin heavy chain expression. Competitive inhibition experiments demonstrated that the elastin-laminin cell surface receptor and its affinity for the VGVAPG peptide sequence on ELP4 molecules are likely involved in the initial SMC contact with the ELP4 modified materials. Elastin-like polypeptides show promise as surface modifiers for candidate scaffolds for engineering contractile vascular tissues.

Electrospun polyurethane nanofiber scaffolds with ciprofloxacin oligomer versus free ciprofloxacin: Effect on drug release and cell attachment

ABSTRACT An electrospun degradable polycarbonate urethane (PCNU) nanofiber scaffold loaded with antibiotic was investigated in terms of antibacterial efficacy and cell compatibility for potential use in gingival tissue engineering. Antimicrobial oligomer (AO), a compound which consists of two molecules of ciprofloxacin (CF) covalently bound via hydrolysable linkages to triethylene glycol (TEG), was incorporated via a one‐step blend electrospinning process using a single solvent system at 7 and 15% w/w equivalent CF with respect to the PCNU. The oligomeric form of the drug was used to overcome the challenge of drug aggregation and burst release when antibiotics are incorporated as free drug. Electrospinning parameters were optimized to obtain scaffolds with similar alignment and fiber diameter to non‐drug loaded fibers. AO that diffused from the fibers was hydrolysed to release CF slowly and in a linear manner over the duration of the study, whereas scaffolds with CF at the same concentration but in free form showed a burst release within 1 h with no further release throughout the study duration. Human gingival fibroblast (HGF) adhesion and spreading was dependent on the concentration and form the CF was loaded (AO vs. free CF), which was attributed in part to differences in scaffold surface chemistry. Surface segregation of AO was quantified using surface‐resolved X‐ray photoelectron spectroscopy (XPS). These findings are encouraging and support further investigation for the use of AO as a means of attenuating the rapid release of drug loaded into nanofibers. The study also demonstrates through quantitative measures that drug additives have the potential to surface‐locate without phase separating from the fibers, leading to fast dissolution and differential fibroblast cell attachment.

Fibronectin adsorption on surface-modified polyetherurethanes and their differentiated effect on specific blood elements related to inflammatory and clotting processes

After the introduction of a medical device into the body, adhesive proteins such as fibronectin (Fn) will adsorb to the surface of the biomaterial. Monocytes (MCs) will interact with these adsorbed proteins, and adopt either a proinflammatory and/or prowound healing phenotype, thereby influencing many blood interaction events including thrombogenesis. In this work, Fn adsorption as well as subsequent MC response and thrombus formation were investigated on two surfaces-modified polyetherurethanes (PEUs) using different surface modifiers: an anionic/dihydroxyl oligomeric (ADO) additive, known to enable cell adhesion, and a fluorinated polypropylene oxide oligomer (PPO), known to reduce platelet adhesion. Results indicated that at 24 h of MC culture, PEU-ADO and PEU-PPO promoted an anti-inflammatory character relative to the base PEU. Longer clotting times, based on a free hemoglobin assay, were also found on the two surface-modified PEUs relative to the native one, suggesting their potential for the reduction of thrombus formation. In presence of a Fn monolayer, the surface-modified PEUs conserved a lower thrombogenic character than the base PEU, and was however significantly decreased when compared to prior protein adsorption. Furthermore, Fn coatings increased the MC production levels of tumor necrosis factor-α and interleukin-10 at 24 h, while not affecting the anti-inflammatory effect of the modifications relative to the base PEU. This finding was most prominent on PEU-PPO, suggesting that the interaction of the adsorbed Fn with blood cells was different for the two additives. Hence, the results highlighted differentiating effects of Fn adsorption on specific blood activating processes related to inflammatory and thrombotic responses.

Influence of ciprofloxacin-based additives on the hydrolysis of nanofiber polyurethane membranes: Influence of ciprofloxacin-based additives

A degradable polycarbonate urethane (PCNU) and an antimicrobial oligomer (AO) were used to generate anti-infective nanofiber scaffolds through blend electrospinning. The AO consists of two molecules of ciprofloxacin (CF) bound through hydrolysable linkages to triethylene glycol. The membranes were conceived for use as tissue engineering scaffolds for the regeneration of soft tissues for the periodontium, where there would be a need for a local dose of antibiotic to the periodontal space as the scaffold degrades in order to prevent biomaterial-associated infection. Scaffolds were made using AO at 7 and 15% w/w equivalent CF, and compared to scaffolds with 15% w/w CF (with HCl counterion). AO was hydrolyzed and released CF continuously over 28 days, while the 15% w/w CF HCl scaffolds showed a burst release within hours, with no subsequent release in the subsequent 28 day period. Released CF from both the AO and CF HCl scaffolds had a similar minimum inhibitory concentration to that of off-the-shelf CF. Interestingly, the introduction of drug in either form (AO or CF HCl) was found to increase the hydrolytic stability of the electrospun degradable PCNU scaffold matrix itself. The alteration of hydrolysis kinetics was attributed to changes in the hydrogen bonding character and microstructure within the scaffolds, introduced by the presence of CF. This study has revealed that in generating in situ drug release systems, the secondary effects of the added drug on the degradation properties of the polymeric carriers must be considered, particularly for systems that act dually as tissue engineering scaffolds.

Synthesis and characterization of Ciprofloxacin-containing divinyl oligomers and assessment of their biodegradation in simulated salivary esterase

OBJECTIVE Two leading causes contributing to dental restoration replacement are the marginal breakdown at the composite/dentin interface and secondary caries mediated by bacteria. The objective of the present study was to synthesize oligomers which incorporated enhanced bio-stability but would also be able to generate antimicrobial function if they underwent degradation. METHODS Stability was incorporated into the oligomers by generating structural features that would physically hinder the availability of hydrolytically sensitive groups in the oligomers. As a proof-of concept for the antibacterial feature, antimicrobial function was achieved by covalently incorporating Ciprofloxacin (CF) into the backbone of cross-linking divinyl oligomers (referred to as EDV and HLH-CFPEG). The hydrolytic stability of the oligomers was studied in simulated human salivary esterase and compared to the commercial monomer 2,2-bis[4(2-hydroxy-3-methacryloxypropoxy)-phenyl]propane (BisGMA). RESULTS Both drug oligomers were found to be significantly more stable than BisGMA. Upon degradation, both drug oligomers released CF differentially in free form. Polymer synthesis from resin formulations containing 15wt% HLH-CFPEG showed a high degree of vinyl group conversion and gel content, and under hydrolytic conditions showed the release of CF during a 28-day monitoring study period. SIGNIFICANCE HLH-CFPEG can be used in dental resin adhesive systems for local delivery of CF to the marginal interface. Minimizing the growth of Streptococcus mutans at the marginal site can improve longevity by reducing esterase activity derived specifically from S. mutans.

Biodegradation Studies of Novel Fluorinated Di-Vinyl Urethane Monomers and Interaction of Biological Elements with Their Polymerized Films

The monomeric components of resin composites in dental restorative materials are susceptible to hydrolysis in the oral cavity. The main objective of this study was to assess the bio-stability of fluorinated urethane dimethacrylates and determine the nature of fluoro-chemistry interactions with protein and bacterial adhesion (both sources of hydrolytic activity) onto cured resin. Degradation studies were performed in the presence of either albumin (in a mildly alkaline pH) or cholesterol esterase (CE). The surface chemistry of the polymers was assessed by water contact angle measurements, pre- and post- incubation with albumin. Adhesion of Streptococcus mutans to cured resin was investigated. The fluorinated monomers were more stable against degradation when compared to the commercial monomer bisphenol A-diglycidyl methacrylate (BisGMA). While fluorinated monomers showed hydrolytic stability with respect to CE, all fluorinated monomers underwent some degree of degradation with albumin. The fluoro-chemistry did not reduce protein and/or bacterial adhesion onto the surface, however post incubation with albumin, the fluorinated surfaces still presented hydrophobic character as determined by the high contact angle values ranging from 79° to 86°. These monomers could potentially be used to increase the hydrophobicity of polymeric composites and provide a means to moderate esterolytic degradation associated with the monomeric component of the polymers within the oral cavity.