Epsin Mimetic UPI Peptide Delivery Strategies to Improve Endothelization of Vascular Grafts.

Endothelialization of engineered vascular grafts for replacement of small-diameter coronary arteries remains a critical challenge. The ability for an acellular vascular graft to promote endothelial cell (EC) recruitment in the body would be very beneficial. This study investigated epsins as a target since they are involved in internalization of vascular endothelial growth factor receptor 2. Specifically, epsin-mimetic UPI peptides are delivered locally from vascular grafts to block epsin activity and promote endothelialization. The peptide delivery from fibrin coatings allowed for controlled loading and provided a significant improvement in EC attachment, migration, and growth in vitro. The peptides have even more important impacts after grafting into rat abdominal aortae. The peptides prevented graft thrombosis and failure that is observed with a fibrin coating alone. They also modulated the in vivo remodeling. The grafts are able to remodel without the formation of a thick fibrous capsule on the adventitia with the 100 µg mL-1 peptide-loaded condition, and this condition enabled the formation of a functional EC monolayer in the graft lumen after only 1 week. Overall, this study demonstrated that the local delivery of UPI peptides is a promising strategy to improve the performance of vascular grafts.

Peritoneal pre-conditioning impacts long-term vascular graft patency and remodeling.

There are questions about how well small-animal models for tissue-engineered vascular grafts (TEVGs) translate to clinical patients. Most TEVG studies used grafting times ≤6 months where conduits from generally biocompatible materials like poly(ε-caprolactone) (PCL) perform well. However, longer grafting times can result in significant intimal hyperplasia and calcification. This study tests the hypothesis that differences in pro-inflammatory response from pure PCL conduits will be consequential after long-term grafting. It also tests the long-term benefits of a peritoneal pre-implantation strategy on rodent outcomes. Electrospun conduits with and without peritoneal pre-implantation, and with 0 % and 10 % (w/w) collagen/PCL, were grafted into abdominal aortae of rats for 10 months. This study found that viability of control grafts without pre-implantation was reduced unlike prior studies with shorter grafting times, confirming the relevance of this model. Importantly, pre-implanted grafts had a 100 % patency rate. Further, pre-implantation reduced intimal hyperplasia within the graft. Differences in response between pure PCL and collagen/PCL conduits were observed (e.g., fewer CD80+ and CD3+ cells for collagen/PCL), but only pre-implantation had an effect on the overall graft viability. This study demonstrates how long-term grafting in rodent models can better evaluate viability of different TEVGs, and the benefits of the peritoneal pre-implantation step.

Reduced Platelet Adhesion for Blended Electrospun Meshes with Low Amounts of Collagen Type I.

A clinically approved, tissue engineered graft is needed as an alternative for small-diameter artery replacement. Collagen type I is commonly investigated for naturally derived grafts. However, collagen promotes thrombosis, currently requiring a graft pre-seeding step. This study investigates unique impacts of blending low collagen amounts with synthetic polymers on scaffold platelet response, which would allow for viable acellular grafts that can endothelialize in vivo. While platelet adhesion and activation are confirmed to be high with 50% collagen samples, low collagen ratios surprisingly exhibit the opposite, anti-thrombogenic effect. Different platelet interactions in these blended materials can be related to collagen structure. Low collagen ratios show homogenous distribution of the components within individual fibers. Importantly, blended collagen scaffolds exhibit significant differences from gelatin scaffolds, including retaining percentage of collagen after incubation. These findings correlate with functional benefits including better endothelial cell spreading on collagen versus gelatin blended materials. This appears to differ from the current paradigm that processing with harsh solvents will irreversibly denature collagen into less desirable gelatin, but an important distinction is the interaction between collagen and synthetic materials during processing. Overall, excellent anti-thrombogenic properties of low collagen blends and benefits after grafting show promise for this vascular graft strategy.

Peritoneal Pre-conditioning Method for In Vivo Vascular Graft Maturation Utilizing a Porous Pouch.

Tissue-engineered vascular grafts (TEVGs) require strategies to allow graft remodeling but avoid stenosis and loss of graft mechanics. A variety of promising biomaterials and methods to incorporate cells have been tested, but intimal hyperplasia and graft thrombosis are still concerning when grafting in small-diameter arteries. Here, we describe a strategy using the peritoneal cavity as an "in vivo" bioreactor to recruit autologous cells to electrospun conduits, which can improve the in vivo response after aortic grafting. We focus on the methods for a novel hydrogel pouch design to enclose the electrospun conduits that can avoid peritoneal adhesion but still allow infiltration of peritoneal fluid and cells needed to provide benefits when subsequently grafting in the aorta.

Arabidopsis cryptochrome is responsive to Radiofrequency (RF) electromagnetic fields.

How living systems respond to weak electromagnetic fields represents one of the major unsolved challenges in sensory biology. Recent evidence has implicated cryptochrome, an evolutionarily conserved flavoprotein receptor, in magnetic field responses of organisms ranging from plants to migratory birds. However, whether cryptochromes fulfill the criteria to function as biological magnetosensors remains to be established. Currently, theoretical predictions on the underlying mechanism of chemical magnetoreception have been supported by experimental observations that exposure to radiofrequency (RF) in the MHz range disrupt bird orientation and mammalian cellular respiration. Here we show that, in keeping with certain quantum physical hypotheses, a weak 7 MHz radiofrequency magnetic field significantly reduces the biological responsivity to blue light of the cryptochrome receptor cry1 in Arabidopsis seedlings. Using an in vivo phosphorylation assay that specifically detects activated cryptochrome, we demonstrate that RF exposure reduces conformational changes associated with biological activity. RF exposure furthermore alters cryptochrome-dependent plant growth responses and gene expression to a degree consistent with theoretical predictions. To our knowledge this represents the first demonstration of a biological receptor responding to RF exposure, providing important new implications for magnetosensing as well as possible future applications in biotechnology and medicine.

Design and characterization of a porous pouch to prevent peritoneal adhesions during in vivo vascular graft maturation.

Vein grafts for coronary artery bypass are not available in more than 30% of patients due to prior use or systemic vascular diseases. Tissue engineered vascular grafts (TEVGs) have shown promise, but intimal hyperplasia and graft thrombosis are still concerns when grafted in small-diameter arteries. In this study, we utilized the peritoneal cavity as an "in vivo" bioreactor to recruit autologous cells to electrospun conduits enclosed within porous pouches to improve the response after grafting. Specifically, we designed a new poly (ethylene glycol)-based pouch to avoid adhesion to the peritoneal wall and still allow the necessary peritoneal fluid to reach the enclosed conduit. The pouch mechanics in compression and bending were determined through experiments and finite element simulations to optimize the pouch design. This included poly (ethylene glycol) concentration, pore density, and pouch size. We demonstrated that the optimized pouch was able to withstand the estimated forces applied in the rat peritoneal cavity and it allowed maturation of the enclosed electrospun conduit. This pouch significantly reduced peritoneal adhesion formation compared to polytetrafluoroethylene pouches that have been used previously, which overcomes this potential limitation to clinical translation. After aortic grafting of pre-conditioned conduits, patent grafts with limited intimal hyperplasia were observed. Overall, this study demonstrated a new pouch design that allows the in vivo bioreactor strategy to be used for vascular tissue engineering without the potential side effect of peritoneal adhesion formation.

Improvement of PDMS surface biocompatibility is limited by the duration of oxygen plasma treatment.

The recent study focused on the improvement of polydimethylsiloxane (PDMS) surface biocompatibility as the most commonly used biomaterial in maxillofacial prostheses for intraoral defects. Biocompatibility enhances tissue-prosthesis integration to prevent implant dislocation; to evaluate the parameter the study conducted at different times of oxygen plasma exposure. Scanning electron microscopy, contact angle measurement, atomic force microscopy and above all, cell cultivation-as a crucial factor in biocompatibility-carried out to investigate the samples' characteristics. An improved PDMS biocompatibility is expected; referring to the fact that an "optimal range"-not necessarily the maximum values-of surface hydrophilicity and roughness could induce an enhanced cell attachment on the PDMS surface, an "optimum time" of O2 plasma exposure is required to meet this goal. Considering the O2 plasma setup items, the ratio of PDMS components and fabrication process in the current survey, 2.5-min O2 plasma exposure well suited to PDMS surface cell adhesion.

Rapid generation of three-dimensional microchannels for vascularization using a subtractive printing technique.

The development of tissue-engineered products has been limited by lack of a perfused microvasculature that delivers nutrients and maintains cell viability. Current strategies to promote vascularization such as additive three-dimensional printing techniques have limitations. This study validates the use of an ultra-fast laser subtractive printing technique to generate capillary-sized channels in hydrogels prepopulated with cells by demonstrating cell viability relative to the photodisrupted channels in the gel. The system can move the focal spot laterally in the gel at a rate of 2500 mm/s by using a galvanometric scanner to raster the in plane focal spot. A Galilean telescope allows z-axis movement. Blended hydrogels of polyethylene glycol and collagen with a range of optical clarities, mechanical properties and swelling behavior were tested to demonstrate that the subtractive printing process for writing vascular channels is compatible with all of the blended hydrogels tested. Channel width and patterns were controlled by adjusting the laser energy and focal spot positioning, respectively. After treatment, high cell viability was observed at distances greater than or equal to 18 µm from the fabricated channels. Overall, this study demonstrates a flexible technique that has the potential to rapidly generate channels in tissue-engineered constructs.