Glycocalyx dynamics and the inflammatory response of genetically modified porcine endothelial cells.

Xenotransplantation is a promising approach to reduce organ shortage, while genetic modification of donor pigs has significantly decreased the immunogenic burden of xenotransplants, organ rejection is still a hurdle. Genetically modified pig organs are used in xenotransplantation research, and the first clinical pig-to-human heart transplantation was performed in 2022. However, the impact of genetic modification has not been investigated on a cellular level yet. Endothelial cells (EC) and their sugar-rich surface known as the glycocalyx are the first barrier encountering the recipient's immune system, making them a target for rejection. We have previously shown that wild type venous but not arterial EC were protected against heparan sulfate (HS) shedding after activation with human serum or human tumor necrosis factor alpha (TNF𝛼). Using a 2D microfluidic system we investigated the glycocalyx dynamics of genetically modified porcine arterial and venous EC (Gal𝛼1,3 Gal knock-out, transgenic for human CD46 and thrombomodulin, GTKO/hCD46/hTM) after activation with human serum or human TNF𝛼. Interestingly, we observed that GTKO/hCD46/hTM arterial cells, additionally to venous cells, do not shed HS. Unscathed HS on GTKO/hCD46/hTM EC correlated with reduced complement deposition, suggesting that protection against complement activation contributes to maintaining an intact glycocalyx layer on arterial EC. This protection was lost on GTKO/hCD46/hTM cells after simultaneous perfusion with human serum and human TNF𝛼. HS shedding on arterial cells and increased complement deposition on both arterial and venous cells was observed. These findings suggest that GTKO/hCD46/hTM EC revert to a proinflammatory phenotype in an inflammatory xenotransplantation setting, potentially favoring transplant rejection.

[Effects of material interfaces on orientation and function of fibrinogen].

Fibrinogen (Fg) in human plasma plays an important role in hemostasis, vascular repair and tissue integrity. The surface chemistry of extracellular matrix or biological materials affects the orientation and distribution of Fg, and changes the exposure of integrin binding sites, thereby affecting its adhesion function to platelets. Here, the quantity, morphology and side chain exposure of Fg adsorbed on hydrophilic, hydrophobic and avidin surfaces were measured by atomic force microscopy (AFM) and flow cytometry (FCM), then the rolling behavior of platelets on Fg was observed through a parallel plate flow chamber system. Our results show that the hydrophobic surface leads to a large amount of cross-linking and aggregation of Fg, while the hydrophilic surface reduces the adsorption and accumulation of Fg while causing the exposure and spreading of the α chain on Fg and further mediating the adhesion of platelets. Fg immobilized by avidin / biotin on hydrophilic surface can maintain the monomer state, avoid over exposure and stretching of α chain, and bind to the platelets activated by the A1 domain of von Willebrand factor instead of inactivated platelets. This study would be helpful for improving the blood compatibility of implant biomaterials and reasonable experimental design of coagulation in vitro.

ICE-Derived Left Atrial and Left Ventricular Endocardial and Myocardial Speckle Tracking Strain Patterns in Atrial Fibrillation at the Time of Radiofrequency Ablation.

Objectives: Intracardiac echocardiography(ICE) has excellent imaging resolution and border recognition which increase strain measurement accuracy. We hypothesized that left atrial(LA) substrate and functional impairment can be detected by measuring LA strain deformation in patients with persistent and paroxysmal atrial fibrillation(AF), as compared to those with no AF. Strain deformation changes in LA and left ventricle(LV) can also be assessed post-ablation to determine its effect. Methods: ICE-derived speckle tracking strain(STS) was prospectively performed in 96 patients, including 62 patients with AF(31 persistent and 31 paroxysmal AF) pre-/post-ablation, and 34 patients with no AF. We measured major strain parameters including longitudinal segmental(endo/myocardial) "average peak overall strain of all segments"(PkAll), peak strain rate(SR),and different time-to-peak strain in LA and LV images. Results: At baseline, persistent AF patients had significantly lower(p<0.01) LA endocardial(4.3±2.5 vs. 20.3±8.9 and 25.5±12.9 %) and myocardial PkAll(4.4±2.6 vs. 15.7±7.2 and 20.9±9.2 %), endocardial(0.9±0.4 vs. 1.8±0.7 and 2.2±0.6 1/s) and myocardial peak SR(0.7±0.4 vs. 1.5±0.6 and 1.9±0.5 1/s), as compared to paroxysmal AF and no AF patients. After successful ablation, endo-/myocardial LA PkAll and peak SR were significantly improved, most dramatically in patients with persistent AF. LV endocardial/myocardial strain and SR also improved in AF patients post-ablation. Conclusion: LA longitudinal strain(%)/SR(1/s) parameters in AF patients are more abnormal than those with no AF, suggesting LA substrate/functional damage. AF ablation improved LA strains/SR but with values in paroxysmal > persistent AF suggesting background LA damage in persistent AF.