Study of the in vitro stress culture of New Zealand rabbit carotid artery
FU Haiyang1 LI Min1,2 YU Hao1,2 FU Bufang1 WANG Zhaoxu1
1.Institute of Medical Devices Control, National Institutes for Food and Drug Control, Beijing 102629, China;
2.College of Life Science, Yantai University, Shandong Province, Yantai 264000, China
Abstract:Objective To study the changes of vascular structure in vitro stress culture of carotid artery in New Zealand rabbits by adjusting the parameters of fluid flow, flow rate and periodic pulsation. Methods Carotid arteries of SPF New Zealand rabbits were put in a bioreactor to establish the vessel culture model in vitro. There were four groups, the static group was static culture, and the dynamic groups (group 80, group 50 and group 25) were maintained under the same operating displacement and frequency of pulse pump, and the flow rates of culture medium were 80, 50 and 25 ml/min, respectively, to maintain the stress state culture. After the culture, the changes of vascular structure were evaluated by HE, Masson and EVG staining. Results In static group, vascular endothelial cells remained intact, smooth muscle cells were arranged in disorder, elastic fibers contracted sharply, and collagen fibers remained basically unchanged at 15 days. In group 80, elastic fibers did not change significantly, and endothelial cells could not be seen,and the smooth muscle in the 8th day was more orderly than that in the 4th day, and collagen fibers decreased significantly in the 8th day. In group 50, endothelial cells were not visible, smooth muscle cells were arranged in order, elastic fibers became loose and flat at the 15th days, and vascular collagen fibers decreased significantly. In group 25, the changes of elastic fibers and collagen fibers were milder than those in group 50. Conclusion Under the condition of simulating various parameters in vivo, the dynamic culture of blood vessels in vitro is helpful to maintain the physiological structure of blood vessels. The difficulty of culture is to prevent pollution and supplement collagen. Under the condition of high stress, vascular endothelial cells will be damaged, resulting in the proliferation of smooth muscle cells and the more serious loss of collagen fibers.
付海洋1 李 敏1,2 于浩1,2 付步芳1 王召旭1. 新西兰家兔颈动脉体外应力培养研究[J]. 中国医药导报, 2021, 18(31): 4-6,16.
FU Haiyang1 LI Min1,2 YU Hao1,2 FU Bufang1 WANG Zhaoxu1. Study of the in vitro stress culture of New Zealand rabbit carotid artery. 中国医药导报, 2021, 18(31): 4-6,16.
[1] 杨广鑫,栾景源.生物可降解金属血管支架研究进展[J].中国微创外科杂志,2018,209(8):753-757,760.
[2] 郑玉峰,杨宏韬. 血管支架用可降解金属研究进展[J].金属学报,2017,53(10):1227-1237.
[3] 毛岳峰,朱晒红,王国慧,等. 聚乳酸在血管支架中的应用及研究进展[J].国际生物医学工程杂志,2008,(6):369-374.
[4] Soares JS,Moore JE Jr. Biomechanical Challenges to Polymeric Biodegradable Stents [J]. Ann Biomed Eng,2016,44(2):560-579.
[5] Wang X,Sun Q,Pei J. Microfluidic-Based 3D Engineered Microvascular Networks and Their Applications in Vascularized Microtumor Models [J]. Micromachines(Basel),2018,9(10):493.
[6] Menon NV,Tay HM,Wee SN,et al. Micro-engineered perfusable 3D vasculatures for cardiovascular diseases [J]. Lab Chip,2017,17(17):2960-2968.
[7] Atchison L,Abutaleb NO,Snyder-Mounts E,et al. iPSC-Derived Endothelial Cells Affect Vascular Function in a Tissue-Engineered Blood Vessel Model of Hutchinson-Gilford Progeria Syndrome [J]. Stem Cell Reports,2020, 14(2):325-337.
[8] Scicchitano P,Cortese F,Gesualdo M,et al. The role of endothelial dysfunction and oxidative stress in cerebrovascular diseases [J]. Free Radic Res,2019,53(6):579-595.
[9] van Duinen V,van den Heuvel A,Trietsch SJ,et al. 96 perfusable blood vessels to study vascular permeability in vitro [J]. Sci Rep,2017,7(1):18071.
[10] Gong MM,Lugo-Cintron KM,White BR,et al. Human organotypic lymphatic vessel model elucidates microenvironment-dependent signaling and barrier function [J]. Biomaterials,2019,214:119225.
[11] Bardy N,Karillon GJ,Merval R,et al. Differential effects of pressure and flow on DNA and protein synthesis and on fibronectin expression by arteries in a novel organ culture system [J]. Circ Res,1995,77(4):684-694.
[12] 刘波,姜宗来,张炎,等.血管体外应力培养系统:一种新的血管生物力学实验模型[J].医用生物力学,2001, 16(4):225-230.
[13] 张海燕.血管支架致血管再狭窄的体外实验研究[D].成都:西南交通大学,2006.
[14] 刘艳秋,黄楠,邹远文,等.一种血管生物反应器相关参数的量化分析[J].西南交通大学学报,2012,47(1):139-143.
[15] Post A,Diaz-Rodriguez P,Balouch B,et al. Elucidating the role of graft compliance mismatch on intimal hyperplasia using an ex vivo organ culture model [J]. Acta Biomater,2019,89:84-94.
[16] Daum R,Visser D,Wild C,et al. Fibronectin Adsorption on Electrospun Synthetic Vascular Grafts Attracts Endothelial Progenitor Cells and Promotes Endothelialization in Dynamic In Vitro Culture [J]. Cells,2020,9(3):778.
[17] Edmunds LH Jr. The Annals of Thoracic Surgery [J]. Ann Thorac Surg,2014,97(1 Suppl):S16-S21.
[18] 邱雅慧.血管内皮细胞的功能以及损伤修复与动脉粥样硬化[J].中国组织工程研究与临床康复,2007,266(10):1927-1929,1933.
[19] 张晓玉,张修彦,詹纯列,等.SP级新西兰兔脏器参数、血液指标、心率、颈动脉血压、心室压的测定与比较[J].中国比较医学杂志,2015(8):37-43.
[20] 刘艳秋,黄楠,毕振飞,等.兔血管内皮细胞在体外应力培养前后的形态学分析[J].稀有金属材料与工程,2014, 43(S1):382-386.
[21] 刘艳春,姜宗来,刘波,等.低切应力对体外培养动脉的平滑肌细胞增殖和凋亡的影响[J].医用生物力学,2002, 17(4):198-202.
[22] 齐帜,谢英.应力致血管平滑肌细胞生长的研究进展[J].武警医学,2008,19(5):470-472.
[23] Atchison L,Zhang H,Cao K,et al. A tissue engineered blood vessel model of Hutchinson-Gilford Progeria syndrome using human iPSC-derived smooth muscle cells [J]. Sci Rep,2017(7),8168-8179.
[24] 陈嵘,程方荣.论血管的力学性质[J].河南医药信息,2001(8):32-33.
[25] Zhang X,Bishawi M,Zhang G,et al. Modeling early stage atherosclerosis in a primary human vascular microphysiological system [J]. Nat Commun,2020,11(1):5426.