Mallis Panagiotis, Sokolis Dimitrios P, Katsimpoulas Michalis, Kostakis Alkiviadis, Stavropoulos-Giokas Catherine, Michalopoulos Efstathios
Hellenic Cord Blood Bank, Biomedical Research Foundation Academy of Athens, 4 Soranou Ephessiou Street, 115 27 Athens, Greece.
Laboratory of Biomechanics, Center for Experimental Surgery, Biomedical Research Foundation Academy of Athens, 4 Soranou Ephessiou Street, 115 27 Athens, Greece.
Bioengineering (Basel). 2021 Aug 27;8(9):118. doi: 10.3390/bioengineering8090118.
The development of functional bioengineered small-diameter vascular grafts (SDVGs), represents a major challenge of tissue engineering. This study aimed to evaluate the repopulation efficacy of biological vessels, utilizing the cord blood platelet lysate (CBPL).
Human umbilical arteries (hUAs, = 10) were submitted to decellularization. Then, an evaluation of decellularized hUAs, involving histological, biochemical and biomechanical analysis, was performed. Wharton's Jelly (WJ) Mesenchymal Stromal Cells (MSCs) were isolated and characterized for their properties. Then, WJ-MSCs (1.5 × 10 cells) were seeded on decellularized hUAs ( = 5) and cultivated with (Group A) or without the presence of the CBPL, (Group B) for 30 days. Histological analysis involving immunohistochemistry (against Ki67, for determination of cell proliferation) and indirect immunofluorescence (against activated MAP kinase, additional marker for cell growth and proliferation) was performed.
The decellularized hUAs retained their initial vessel's properties, in terms of key-specific proteins, the biochemical and biomechanical characteristics were preserved. The evaluation of the repopulation process indicated a more uniform distribution of WJ-MSCs in group A compared to group B. The repopulated vascular grafts of group B were characterized by greater Ki67 and MAP kinase expression compared to group A.
The results of this study indicated that the CBPL may improve the repopulation efficacy, thus bringing the biological SDVGs one step closer to clinical application.
功能性生物工程小口径血管移植物（SDVGs）的开发是组织工程的一项重大挑战。本研究旨在评估利用脐带血血小板裂解物（CBPL）对生物血管的再填充效果。
对人脐动脉（hUAs，n = 10）进行去细胞处理。然后，对去细胞化的hUAs进行组织学、生化和生物力学分析评估。分离并鉴定了华通氏胶（WJ）间充质基质细胞（MSCs）的特性。然后，将WJ-MSCs（1.5×10个细胞）接种到去细胞化的hUAs（n = 5）上，并在有（A组）或无CBPL存在的情况下（B组）培养30天。进行了包括免疫组织化学（针对Ki67，用于测定细胞增殖）和间接免疫荧光（针对活化的MAP激酶，细胞生长和增殖的额外标志物）的组织学分析。
去细胞化的hUAs在关键特异性蛋白方面保留了其初始血管的特性，生化和生物力学特性得以保留。再填充过程的评估表明，与B组相比，A组中WJ-MSCs的分布更均匀。与A组相比，B组再填充的血管移植物具有更高的Ki67和MAP激酶表达。
本研究结果表明，CBPL可能提高再填充效果，从而使生物SDVGs向临床应用迈进了一步。
