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自体耳软骨细胞用于人工表面衬里的可行性研究。

Use of autologous auricular chondrocytes for lining artificial surfaces: a feasibility study.

作者信息

Scott-Burden Timothy, Bosley Jennifer P, Rosenstrauch Doreen, Henderson Kimberly D, Clubb Fred J, Eichstaedt Harald C, Eya Kazuhiro, Gregoric Igor, Myers Timothy J, Radovancevic Branislav, Frazier O H

机构信息

Cardiovascular Surgical Research Laboratories, Texas Heart Institute at St. Luke's Episcopal Hospital, Houston, USA.

出版信息

Ann Thorac Surg. 2002 May;73(5):1528-33. doi: 10.1016/s0003-4975(01)03600-1.

Abstract

BACKGROUND

Auricular elastic cartilage is a potential source of autologous cells for lining the luminal surfaces of cardiovascular prostheses. We tested this potential in vitro and in vivo using a left ventricular assist device (LVAD) and a calf model.

METHODS

In vitro, auricular cartilage was harvested from the anesthetized ear of a calf, isolated, and cultured on tissue culture dishes. Primary chondrocytes were typed by immunocytochemistry, transferred into culture media, passaged twice, and seeded onto the blood-contacting luminal surfaces of four LVADs (HeartMate; Thoratec Corporation, Woburn, MA). Seeded cell linings were preconditioned under simulated flow conditions to promote cell adhesion to luminal surfaces. Seeding efficiency and cumulative cell loss under flow conditions were quantitated. In vivo, one of the four autologous chondrocyte-lined and preconditioned LVADs was implanted into the tissue-donor calf; run for 7 days; explanted; and evaluated grossly, by scanning electron microscopy, and by transmission electron microscopy.

RESULTS

The efficiency of seeding chondrocytes onto the luminal surfaces of the four LVADs was 95.11% +/- 4.23% (n = 4). Cumulative cell loss during preconditioning under flow conditions in vitro did not exceed 12% (n = 4). After 7 days of in vivo implantation, the luminal surfaces of the implanted LVAD demonstrated an intact, strongly adherent cellular lining.

CONCLUSIONS

Auricular elastic cartilage is a ready and easily accessible source of chondrocytes whose ability to produce collagen II and other important extracellular matrix constituents allows them to adhere strongly to the luminal surfaces of LVADs. The simple method of isolating and expanding auricular chondrocytes presented here could be used to provide strongly adherent autologous cell linings for LVADs and other cardiovascular devices. If and when chondrocytes can be genetically engineered to produce antithrombogenic factors and then used to line the luminal surfaces of LVADs or other cardiovascular prostheses, they may be able to improve the hemocompatibility of the blood-biomaterial interface in such devices. Our successful feasibility study in a calf model warrants further studies of this concept in vivo.

摘要

背景

耳廓弹性软骨是用于心血管假体腔面内衬的自体细胞的潜在来源。我们使用左心室辅助装置(LVAD)和小牛模型在体外和体内测试了这种潜力。

方法

在体外,从小牛麻醉的耳朵中采集耳廓软骨,分离并在组织培养皿中培养。通过免疫细胞化学对原代软骨细胞进行分型,转移到培养基中,传代两次,然后接种到四个LVAD(HeartMate;Thoratec公司,马萨诸塞州沃本)的血液接触腔面上。接种的细胞内衬在模拟流动条件下进行预处理,以促进细胞与腔面的粘附。对流动条件下的接种效率和累积细胞损失进行定量。在体内,将四个自体软骨细胞内衬并预处理的LVAD之一植入组织供体小牛体内;运行7天;取出;并通过大体观察、扫描电子显微镜和透射电子显微镜进行评估。

结果

将软骨细胞接种到四个LVAD腔面上的效率为95.11%±4.23%(n = 4)。体外流动条件下预处理期间的累积细胞损失不超过12%(n = 4)。体内植入7天后,植入的LVAD腔面显示出完整、牢固粘附的细胞内衬。

结论

耳廓弹性软骨是软骨细胞现成且易于获取的来源,其产生II型胶原蛋白和其他重要细胞外基质成分的能力使其能够牢固粘附于LVAD的腔面。本文介绍的分离和扩增耳廓软骨细胞的简单方法可用于为LVAD和其他心血管装置提供牢固粘附的自体细胞内衬。如果并且当软骨细胞能够通过基因工程产生抗血栓形成因子,然后用于内衬LVAD或其他心血管假体的腔面时,它们可能能够改善此类装置中血液-生物材料界面的血液相容性。我们在小牛模型中成功的可行性研究值得对这一概念进行进一步的体内研究。

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