{"title":"Sphingosine-1-Phosphate (S1P) in Whole Liver Recellularization Improves Endothelization of Acellular Liver Scaffold.","authors":"Usha Yadav, Chandra J Yadav, Sadia Afrin, Jun-Yeong Lee, Jihad Kamel, Kyung-Mee Park","doi":"10.1021/acsbiomaterials.5c00411","DOIUrl":null,"url":null,"abstract":"<p><p>Endothelialization is crucial for tissue bioengineering, particularly in developing functional blood vessel linings to ensure proper vascularization. Effective re-endothelialization of the vasculature in bioengineered organs is challenging, often leading to blood coagulation and hindering successful engraftment. Endothelial cell proliferation, migration, and angiogenesis are essential processes for constructing functional and vascularized bioengineered organs. Sphingosine-1-phosphate (S1P), a low-molecular-weight phospholipid mediator, regulates various biological activities in endothelial cells including survival, proliferation, and cell barrier integrity. In this study, we present a novel approach to enhance the re-endothelialization of decellularized rat liver scaffolds by seeding human umbilical vein endothelial cells (HUVECs) in the presence of S1P, aiming to bioengineer a fully endothelialized liver. Initially, we validated the effects of S1P on HUVECs in a 2D cell culture system, confirming that S1P significantly promotes endothelial functions. Following this validation, we seeded HUVECs in the presence of S1P into decellularized rat liver scaffolds via the portal vein. The seeded liver was maintained in the bioreactor and perfused with medium supplemented with S1P for 7 days. The efficacy of S1P on liver scaffolds was evaluated through the longitudinal monitoring of cell proliferation using the resazurin reduction assay, indicating higher cell proliferation in the constructs. Further characterization through histological and molecular analyses demonstrated efficient coverage of vessels of re-endothelialized scaffolds maintaining their function. The antithrombotic effect of the fully endothelialized scaffold was assessed via ex vivo whole-blood perfusion. Our results indicate that S1P is a key regulator of endothelialization processes, promoting HUVECs proliferation and survival and facilitating the formation of a functional endothelial layer on the vascular structure of re-endothelialized liver scaffold.</p>","PeriodicalId":8,"journal":{"name":"ACS Biomaterials Science & Engineering","volume":" ","pages":""},"PeriodicalIF":5.4000,"publicationDate":"2025-06-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"ACS Biomaterials Science & Engineering","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1021/acsbiomaterials.5c00411","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"MATERIALS SCIENCE, BIOMATERIALS","Score":null,"Total":0}
引用次数: 0
Abstract
Endothelialization is crucial for tissue bioengineering, particularly in developing functional blood vessel linings to ensure proper vascularization. Effective re-endothelialization of the vasculature in bioengineered organs is challenging, often leading to blood coagulation and hindering successful engraftment. Endothelial cell proliferation, migration, and angiogenesis are essential processes for constructing functional and vascularized bioengineered organs. Sphingosine-1-phosphate (S1P), a low-molecular-weight phospholipid mediator, regulates various biological activities in endothelial cells including survival, proliferation, and cell barrier integrity. In this study, we present a novel approach to enhance the re-endothelialization of decellularized rat liver scaffolds by seeding human umbilical vein endothelial cells (HUVECs) in the presence of S1P, aiming to bioengineer a fully endothelialized liver. Initially, we validated the effects of S1P on HUVECs in a 2D cell culture system, confirming that S1P significantly promotes endothelial functions. Following this validation, we seeded HUVECs in the presence of S1P into decellularized rat liver scaffolds via the portal vein. The seeded liver was maintained in the bioreactor and perfused with medium supplemented with S1P for 7 days. The efficacy of S1P on liver scaffolds was evaluated through the longitudinal monitoring of cell proliferation using the resazurin reduction assay, indicating higher cell proliferation in the constructs. Further characterization through histological and molecular analyses demonstrated efficient coverage of vessels of re-endothelialized scaffolds maintaining their function. The antithrombotic effect of the fully endothelialized scaffold was assessed via ex vivo whole-blood perfusion. Our results indicate that S1P is a key regulator of endothelialization processes, promoting HUVECs proliferation and survival and facilitating the formation of a functional endothelial layer on the vascular structure of re-endothelialized liver scaffold.
期刊介绍:
ACS Biomaterials Science & Engineering is the leading journal in the field of biomaterials, serving as an international forum for publishing cutting-edge research and innovative ideas on a broad range of topics:
Applications and Health – implantable tissues and devices, prosthesis, health risks, toxicology
Bio-interactions and Bio-compatibility – material-biology interactions, chemical/morphological/structural communication, mechanobiology, signaling and biological responses, immuno-engineering, calcification, coatings, corrosion and degradation of biomaterials and devices, biophysical regulation of cell functions
Characterization, Synthesis, and Modification – new biomaterials, bioinspired and biomimetic approaches to biomaterials, exploiting structural hierarchy and architectural control, combinatorial strategies for biomaterials discovery, genetic biomaterials design, synthetic biology, new composite systems, bionics, polymer synthesis
Controlled Release and Delivery Systems – biomaterial-based drug and gene delivery, bio-responsive delivery of regulatory molecules, pharmaceutical engineering
Healthcare Advances – clinical translation, regulatory issues, patient safety, emerging trends
Imaging and Diagnostics – imaging agents and probes, theranostics, biosensors, monitoring
Manufacturing and Technology – 3D printing, inks, organ-on-a-chip, bioreactor/perfusion systems, microdevices, BioMEMS, optics and electronics interfaces with biomaterials, systems integration
Modeling and Informatics Tools – scaling methods to guide biomaterial design, predictive algorithms for structure-function, biomechanics, integrating bioinformatics with biomaterials discovery, metabolomics in the context of biomaterials
Tissue Engineering and Regenerative Medicine – basic and applied studies, cell therapies, scaffolds, vascularization, bioartificial organs, transplantation and functionality, cellular agriculture