cardiac tissue engineering strategies have the potential to regenerate functional myocardium following myocardial infarction. In this study, we utilized novel electrospun fibrin microfiber sheets of different stiffnesses (50.0 ± 11.2 kPa and 90.0 ± 16.4 kPa) to engineer biomimetic models of vascularized cardiac tissues. We characterized tissue assembly, electrophysiology, and contractility of neonatal rat ventricular cardiomyocytes (NRVCMs) cultured on these sheets. NRVCMs cultured on the softer substrates displayed higher conduction velocities (CVs) and improved electrophysiological properties. Human umbilical vein endothelial cells (HUVECs) formed dense networks on the sheets when co-cultured with human adipose-derived stem/stromal cells (hASCs). To achieve vascularized cardiac tissues, we tested various tri-culture protocols of NRVCM:hASC:HUVEC and found that a ratio of 1,500,000:37,500:150,000 cells/cm 2 enabled the formation of robust endothelial networks while retaining statistically identical electrophysiological characteristics to NRVCM-only cultures. Tri-cultures at this ratio on 90 kPa substrates exhibited average CVs of 14 ± 0.6 cm/s, Action Potential Duration (APD)80 and APD30 of 152 ± 11 ms and 71 ± 6 ms, respectively, and maximum capture rate (MCR) of 3.9 ± 0.7 Hz. These data indicate the significant potential of generating densely packed endothelial networks together with electrically integrated cardiac cells in vitro as a physiologic 3D cardiac model.

To identify barriers and facilitators to adopting deep anterior lamellar keratoplasty (DALK) for nonendothelial corneal pathology.Methods: An anonymous survey consisting of 22 multiple choice and free text questions was designed to gather information on demographic factors of surgeons and DALK surgical practices. The survey was emailed to members of the kera-net, a global online corneal surgeon/surgery platform.Results: A total of 100 surgeons completed the survey, most of whom practice in the United States (73%). Most surgeons (89%) reported performing DALK. Surgeons who did not learn DALK during fellowship (34%) tended to be in practice for higher numbers of years (P , 0.001). Surgeons in private practice are more likely to perform DALK versus those in other settings (92.7% vs. 80.8%, P = 0.087). Surgeons performing more corneal surgeries (at least 100 per year) are more likely to perform DALK than those who perform fewer than 100 per year (52% vs. 14%, P = 0.01). Surgeons who perform Descemet membrane endothelial keratoplasty are more likely to perform DALK than those who do not (81.7% vs. 18.3%, P = 0.014). There was also a positive correlation between PK and DALK surgical volumes (Spearman rank correlation coefficient = 0.57, P , 0.001). The main reasons for surgeon preference for DALK over PK were a desire to preserve the endothelium, intraoperative safety, and decreased complications. Longer surgical time and low patient volume were cited as barriers to adoption of DALK.Conclusions: Alterations in DALK technique that reduce surgical time and providing more learning opportunities for DALK might improve adoption.

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