Improved delivery of PLGA microparticles and microparticle-cell scaffolds in clinical needle gauges using modified viscosity formulations.

Polymer microparticles are widely used as acellular drug delivery platforms in regenerative medicine, and have emerging potential as cellular scaffolds for therapeutic cell delivery. In the clinic, PLGA microparticles are typically administered intramuscularly or subcutaneously, with the clinician and clinical application site determining the precise needle gauge used for delivery. Here, we explored the role of needle diameter in microparticle delivery yield, and develop a modified viscosity formulation to improve microparticle delivery across a range of clinically relevant needle diameters. We have identified an optimal biocompatible formulation containing 0.25% pluronic F127 and 0.25% carboxymethyl cellulose, which can increase delivery payload to 520% across needle gauges 21-30G, and note that needle diameter impacts delivery efficacy. We use this formulation to increase the delivery yield of PLGA microparticles, and separately, PLGA-cell scaffolds supporting viable mesenchymal stem cells (MSCs), demonstrating the first in vitro delivery of this cell scaffold system. Together, these results highlight an optimal formulation for the delivery of microparticle and microparticle-cell scaffolds, and illustrate how careful choice of delivery formulation and needle size can dramatically impact delivery payload.

Design of maleimide-functionalised electrodes for covalent attachment of proteins through free surface cysteine groups.

Mixed two-component monolayers on glassy carbon are prepared by electrochemical oxidation of N-(2-aminoethyl)acetamide and mono-N-Boc-hexamethylenediamine in mixed solution. Subsequent N-deprotection, amide coupling and solid-phase synthetic steps lead to electrode-surface functionalisation with maleimide, with controlled partial coverage of this cysteine-binding group at appropriate dilution for covalent immobilisation of a model redox-active protein, cytochrome c, with high coverage (≈7.5 pmol cm(-2) ).

Mucoadhesive polymer films for tissue retraction in laparoscopic surgery: Ex-vivo study on their mechanical properties.

Safe and effective manipulation of soft tissue during laparoscopic procedures can be achieved by the use of mucoadhesive polymer films. A series of novel adhesive polymer films were formulated in house based on either Carbopol or Chitosan modified systems. The mechanical properties of the polymers and their adherence to bowel were evaluated using ex-vivo pig bowel immersed in 37°C water bath and connected to an Instron tensiometer. Young's modulus was 300 kPa for the Carbopol-polymer and 5 kPa for the Chitosan-polymer. The Chitosan-polymer exhibited much larger shear adhesion than its tensile adhesion: 3.4 N vs. 1.2. Both tensile and shear adhesions contributed to the large retraction force (2.6 N) obtained during l polymer-bowel retraction testing. Work of adhesion at the polymer/serosa interface, defined as the area under the force curve, was 64 mJ, which is appreciably larger than that reported with existing polymers. In conclusion, adhesive polymers can stick to the serosal side of the bowel with an adhesive force, which is sufficient to lift the bowel, providing a lower retraction stress than that caused by laparoscopic grasping which induces high localized pressures on the tissue.

The effect of dilute solution properties on poly(vinyl alcohol) films.

Films are commonly prepared by solvent casting and accordingly it is important to investigate the relationship between the properties of the polymeric solution and the resultant solid-state properties of the film. Therefore the objective of this work was to characterise the rheological properties of aqueous solutions of PVA, containing defined co-solvents, and to relate these properties to the resultant mechanical and swelling properties of films cast from these solutions. Casting solutions were prepared using two concentrations of two molecular weights of PVA within a range of cosolvent systems. The solution properties were examined using dilute solution viscometry at 37°C and defined in terms of the intrinsic viscosity and Huggins constant. Solutions of PVA containing each cosolvent exhibited greater intrinsic viscosities (and lower Huggins constants) than comparator aqueous solutions of this polymer. In particular propylene glycol and pyrrolidone solvents when blended with water produced thermodynamically good solvents for PVA whereas water was shown to be a thermodynamically poor solvent for PVA. Films were cast from the above solvent systems and exhibited a wide range of mechanical properties (UTS, % elongation at break and Young's Modulus) and swelling properties, both with no signs of ageing These were dependent on the nature of cosolvent used as the casting solvent. A strong correlation (r>±0.9) was observed between the intrinsic viscosity (and Huggins constant) and the observed values of Young's modulus cast from each solvent system. These observations confirmed the importance of the chemical properties of the cosolvent on the mechanical and swelling properties of films manufactured from these solutions.

Ex vivo comparison of mechanical versus thermal chondroplasty: assessment of tissue effect at the surgical endpoint.

PURPOSE: The purpose of this study was to evaluate tissue effect (tissue removal plus underlying cell death) of two chondroplasty techniques: mechanical debridement (MD) using a rotary shaver blade and thermal chondroplasty using radiofrequency energy (RFE). METHODS: Forty-eight human chondromalacic cartilage samples were treated with either MD or RFE. Pre- and post-treatment arthroscopic images of the cartilage surface were recorded. Samples were incubated with cell viability stain and visualized with confocal laser microscopy to determine tissue effect. Smoothing was quantitated by three surgeons using a visual analog scale (VAS) as well as a subjective rating regarding whether smoothing was "arthroscopically acceptable." RESULTS: Tissue effect at the surgical endpoint of arthroscopically acceptable smoothing was 385 microm for MD versus 236 microm for RFE, a significant difference (P < .0001). Mean post-treatment VAS for MD was 2.8 points less smooth than for RFE (P < .0001). Overall, arthroscopically acceptable smoothing was achieved in 90% of RFE samples compared to 49% of MD samples. CONCLUSIONS: Our results shown that chondroplasty using a RFE probe results in greater smoothing of chondromalacic cartilage in fewer treatment passes and with decreased total tissue effect than MD using a rotary shaver blade. CLINICAL RELEVANCE: If safety and efficacy can be shown in vivo, thermal chondroplasty may represent an alternative for treatment of symptomatic chondromalacia.

An ex vivo thermal chondroplasty model: the association of a char-like layer and underlying cell death.

PURPOSE: The purpose of this study was to evaluate the relation between the char-like layer observed during radiofrequency energy (RFE) treatment of cartilage and the depth of underlying cell death. METHODS: Healthy adult bovine patellae were treated with a monopolar RFE probe ex vivo at generator settings of 20, 30, 40, 50, 60, 80, and 110 in cut mode. The presence or absence of a char-like layer and visual electrical discharge was noted. Treated tissue was incubated with cell viability stain, and the depth of cell death and matrix debridement was measured from confocal laser microscopy images. RESULTS: At generator settings of 60 and above, a char-like layer, electrical discharge, and matrix debridement were consistently observed, and the depth of cell death was significantly less (P < .05) than when these features were not observed (< or =30). Paradoxically, the least depth of cell death did not occur at the lowest generator setting in cut mode. It occurred at a generator setting of 60. An increase in impedance of the system and a decrease in current were also associated with reduced cell death. CONCLUSIONS: In this controlled ex vivo study formation of a char-like layer, visual electrical discharge, increased impedance, and reduced current were associated with less depth of cell death when cartilage was treated with monopolar RFE. CLINICAL RELEVANCE: This study suggests that a char-like layer and electrical discharge during RFE treatment of cartilage may be advantageous because, potentially, these features are associated with less depth of cell death (safety) and greater matrix debridement (efficacy).

Determination of factors influencing tissue effect of thermal chondroplasty: an ex vivo investigation.

PURPOSE: Scientific investigation of thermal chondroplasty using radiofrequency energy (RFE) is confounded by multiple factors associated with the technique. Our purpose was to determine the relative importance of the following factors on tissue effect (depth of tissue debridement plus depth of underlying cell death) of thermal chondroplasty: probe design, generator power setting, speed, force, and number of passes of the probe over treated tissue. We hypothesized the relative importance of these factors would be (from most to least important) power, passes, speed, force, and design. METHODS: Bovine patellae were treated using monopolar RFE. Sample size was based on a 2-level, half-factorial design. Low and high extremes of the factors tested were power setting (50 W v 110 W), passes (1 v 5), speed (3 mm/sec v 10 mm/sec), force (0.15 N v and 0.59 N), and probe design (electrode protrusion 25 microm v 125 microm). Samples were incubated with cell viability stain and examined using confocal laser microscopy to determine tissue effect. Data were analyzed using multiple regression. RESULTS: All factors that were tested significantly influenced tissue effect (P < .05). Power setting had the greatest effect, followed by design, speed, passes, and force. The following interactions of factors were also significant: design and force, power and passes. The optimal configuration resulting in least tissue effect was a power setting of 50 W, electrode protrusion of 25 microm, speed of 10 mm/sec, 1 pass, and 0.15 N of applied force during treatment, which resulted in a predicted tissue effect of 99 +/- 15 microm. CONCLUSIONS: The least tissue effect of thermal chondroplasty was achieved with lower power using a probe with minimal electrode protrusion while performing a rapid, single, lower force pass of the probe over treated tissue. CLINICAL RELEVANCE: Power and probe design have the greatest influence among the factors tested; selecting these parameters preoperatively could control tissue effect.