Influence of advanced wound matrices on observed vacuum pressure during simulated negative pressure wound therapy.

Biomaterials and negative pressure wound therapy (NPWT) are treatment modalities regularly used together to accelerate soft-tissue regeneration. This study evaluated the impact of the design and composition of commercially available collagen-based matrices on the observed vacuum pressure delivered under NPWT using a custom test apparatus. Specifically, testing compared the effect of the commercial products; ovine forestomach matrix (OFM), collagen/oxidized regenerated cellulose (collagen/ORC) and a collagen-based dressing (CWD) on the observed vacuum pressure. OFM resulted in an ∼50% reduction in the observed target vacuum pressure at 75 mmHg and 125 mmHg, however, this effect was mitigated to a ∼0% reduction when fenestrations were introduced into the matrix. Both collagen/ORC and CWD reduced the observed vacuum pressure at 125 mmHg (∼15% and ∼50%, respectively), and this was more dramatic when a lower vacuum pressure of 75 mmHg was delivered (∼20% and ∼75%, respectively). The reduced performance of the reconstituted collagen products is thought to result from the gelling properties of these products that may cause occlusion of the delivered vacuum to the wound bed. These findings highlight the importance of in vitro testing to establish the impact of adjunctive therapies on NPWT, where effective delivery of vacuum pressure is paramount to the efficacy of this therapy.

Evaluation of Tissue Apposition and Seroma Prevention in an Ovine Model of Surgical Dead Space Using a Novel Air-Purged Vacuum Closure System.

Background: Postoperative complications associated with seroma formation resulting from surgical dead space continue to present a challenge in modern surgery. There is an unmet need for new technologies that address surgical dead space as well as prevent seroma formation and associated downstream postoperative complications. Methods: The novel implantable tissue apposition and drainage system ENIVO was developed and tested in a bilateral ovine external abdominal oblique (EAO) resection model of surgical dead space. The ENIVO system is a portable powered pump and wound interface featuring air-purged vacuum closure (APVC) that delivers a sustained level of vacuum pressure (80 and 100 mmHg) to the treatment site with an intermittent burst of sterile filtered air through the implanted wound interface. Seroma area, seroma volume, and drain migration were assessed at postoperative days 7 and 14, and all animals were euthanized at day 28 with gross assessment of treatment efficacy including the presence of residual seroma and tissue apposition. Results: The bilateral model created relatively uniform defects of ~120 cm2 following excision of ~30 to 50 g of EAO muscle. Median seroma area of ENIVO-treated defects was statistically smaller than standard of care (SoC)-treated defects at days 7 and 14. Median seroma volume at 14 days was significantly reduced in ENIVO-treated defects relative to SoC-treated defects [1.3 (IQR 0.0-79.5) mL and 188.5 (IQR 27.6-342.9) mL, respectively]. At postoperative day 28, 40% (n = 4/10) of SoC defects showed a residual seroma, whereas in contrast, none of the ENIVO-treated defects showed signs of a residual seroma. Median tissue apposition scoring was higher in the ENIVO treatment group [3 (IQR 3-3)] compared with the SoC group [3 (IQR 0-3)]. Conclusions: The ENIVO system represents a new approach to dead space management and seroma prevention and was shown to outperform a SoC surgical drain in a challenging large defect model of surgical dead space management and seroma prevention.

Quantitative analysis of biomolecule release from polystyrene-block-polyethylene oxide thin films.

Block copolymers have garnered recent attention due to their ability to contain molecular cargo within nanoscale domains and release said cargo in aqueous environments. However, the release kinetics of cargo from these thin-films has not yet been reported. Knowledge of the release quantities and release profiles of these systems is paramount for applications of these systems. Here, Polystyrene-block-poly(ethylene oxide) (PS-b-PEO) was co-assembled with fluorescein isothiocyanate isomer I-lysozyme (FITC-LZ) and fluorescein isothiocyanate isomer I-TAT (FITC-TAT), such that these molecular cargos arrange within the PEO domains of the thin films. We show that high loading ratios of cargo/PS-b-PEO do not significantly impact the nanostructure of the films; however, a loading limit appears to be present with aggregates of protein forming at the microscale with higher loading ratios. The presence of lysozyme (LZ) within the films was confirmed qualitatively after aqueous exposure through photo-induced force microscopy (PiFM) imaging at the Amide I characteristic peak (∼1650 cm-1). Furthermore, we demonstrate that LZ maintains activity and structure after exposure to the polymer solvent (benzene/methanol/water mix). Finally, we demonstrate quantitatively 20-80 ng cm-2 of cargo is released from these films, depending on the cargo incorporated. We show that the larger molecule lysozyme is released over a longer time than the smaller TAT peptide. Finally, we demonstrate the ability to tune the quantity of cargo released by altering the thickness of the PS-b-PEO thin-films during fabrication.

Mechanical Characterization for Cellular Mechanobiology: Current Trends and Future Prospects.

Accurate mechanical characterization of adherent cells and their substrates is important for understanding the influence of mechanical properties on cells themselves. Recent mechanobiology studies outline the importance of mechanical parameters, such as stress relaxation and strain stiffening on the behavior of cells. Numerous techniques exist for probing mechanical properties and it is vital to understand the benefits of each technique and how they relate to each other. This mini review aims to guide the reader through the toolbox of mechanical characterization techniques by presenting well-established and emerging methods currently used to assess mechanical properties of substrates and cells.

Polystyrene-block-poly(ethylene oxide) Thin Films Fabricated from a Solvent Mixture for the Co-Assembly of Polymers and Proteins.

The co-assembly of peptides and proteins in poly(styrene-block-ethylene oxide) (PS-b-PEO) thin films has proven to be a promising method to fabricate polymer-biomolecule functional materials. Contrary to the covalent immobilization of biomolecules on surfaces, co-assembly presents the opportunity to arrange cargo within thin films, which can be released upon exposure to an aqueous environment. The use of a mixed solvent system ensures the solubilization of hydrophobic polymer as well as the solubilization and protection of the biomolecule cargo. However, to produce largely defect-free films of PS-b-PEO from a solvent mixture containing water is challenging due to the narrow range of solvent miscibility and polymer/protein solubility. This work explores the limits of using a benzene/methanol/water solvent mixture for the production of thin PS-b-PEO films and provides a template for the fabrication optimization of block copolymer thin films in different complex solvent systems. The film quality is analyzed using optical microscopy and atomic force microscopy and correlated to the solvent composition. By adjusting the solvent composition to 80/18.8/1.2 vol % benzene/methanol/water, it was possible to reliably fabricate thin films with less than 1% macroscopic defect surface coverage. Using the optimized solvent composition, we also demonstrate the fabrication of ordered PS-b-PEO films containing lysozyme. Furthermore, we show the release of lysozyme into aqueous media, which highlights the potential use of such films for drug delivery applications.

Polystyrene-block-polyethylene oxide thin films: In vitro cytocompatibility and protein adsorption testing.

Polystyrene-block-polyethylene oxide (PS-b-PEO) coated surfaces have been explored as cell culture substrates in the past decade. However, their cytocompatibility has not been extensively assessed. In this study, the in vitro cytocompatibility of PS-b-PEO was investigated. Cellular morphology, metabolic activity, and viability were evaluated at 1, 3, and 5 days after cell seeding. Viability was greater than 90% throughout the 5 days culture, with abundant cell spreading evident by the formation of prominent F-actin stress fibres. The cytocompatibility study was complemented by the analysis of adsorption of a range of extracellular matrix proteins on PS-b-PEO thin films by quartz crystal microbalance with dissipation. Protein adsorption tests revealed that there was no significant difference in protein adhesion between surfaces with a PEO domain coverage of ≈28%, compared to the homogeneous polystyrene control. The findings demonstrate that PS-b-PEO thin films are cytocompatible and are a favourable surface coating for cell culture studies.

Engineered systems to study the synergistic signaling between integrin-mediated mechanotransduction and growth factors (Review).

While tremendous leaps in knowledge into cellular signaling and control have been achieved over the last few decades, there is still more to learn in how different signaling pathways act synergistically. A better understanding and control of cells in vitro and in vivo is important to enable more successful and safe applications of tissue engineering and stem cell therapy. This review is focused on two central ways cells sense their surroundings, namely, integrin-mediated mechanotransduction and growth factor signaling. Specifically, the authors explore how engineered interfaces have been applied to learn more about these processes, and how these important signaling pathways interact synergistically.