Sabashnikov Anton, Neef Klaus, Chesnokova Vera, Wegener Leonie, Godthardt Kathrin, Scherner Maximilian, Kuhn Elmar W, Deppe Antje-Christin, Lauer Meike, Eghbalzadeh Kaveh, Zeriouh Mohamed, Rahmanian Parwis B, Wippermann Jens, Kuhn-Régnier Ferdinand, Madershahian Navid, Wahlers Thorsten, Weymann Alexander, Choi Yeong-Hoon
Department of Cardiothoracic Surgery, Heart Center, University Hospital Cologne, Cologne, Germany.
Center for Molecular Medicine Cologne, University Hospital Cologne, Cologne, Germany.
J Cardiothorac Surg. 2017 Aug 9;12(1):68. doi: 10.1186/s13019-017-0631-3.
The frequency of circulating endothelial cells (CEC) in patients' peripheral blood can be assessed as a direct marker of endothelial damage. However, conventional enumeration methods are extremely challenging. We developed a novel, automated approach to determine CEC frequencies and tested this method on two groups of patients undergoing conventional (CAVR) versus trans-catheter aortic valve implantation (TAVI).
CEC frequencies were assessed by a flow cytometric approach, including automated pre-enrichment of CD34 positive blood cell subpopulation and isotype controls. The efficacy and reproducibility of the CEC enumeration method was validated by spiking blood samples of healthy control donors with defined numbers of endothelial cells.
CEC frequencies were significantly higher in the TAVI group before (9.8 ± 4.1 vs. 5.5 ± 2.2, p = 0.019) and 1 h after surgery (13.4 ± 5.1 vs. 8.2 ± 4.1, p = 0.030) corresponding to higher Euroscore, STS score in higher risk patients from the TAVI group. Five days after surgery, CEC frequencies became significantly higher in the more invasive CAVR group (39.0 ± 13.0 vs. 14.3 ± 4.4, p < 0.001) compared to minimally invasive TAVI approach.
The new flow cytometric approach might be a robust and reliable method for CEC enumeration. Initial results show that CEC frequency is a valid clinical marker for the assessment of pre-operative risk, invasiveness of surgical procedure and clinical outcome. Further studies are necessary to validate the practical clinical usefulness and the potential superiority compared to conventional markers.
患者外周血中循环内皮细胞（CEC）的频率可作为内皮损伤的直接标志物进行评估。然而，传统的计数方法极具挑战性。我们开发了一种新型的自动化方法来确定CEC频率，并在两组接受传统主动脉瓣置换术（CAVR）与经导管主动脉瓣植入术（TAVI）的患者中对该方法进行了测试。
通过流式细胞术方法评估CEC频率，包括对CD34阳性血细胞亚群进行自动预富集和同型对照。通过向健康对照供者的血样中加入确定数量的内皮细胞，验证了CEC计数方法的有效性和可重复性。
TAVI组术前（9.8±4.1对5.5±2.2，p = 0.019）和术后1小时（13.4±5.1对8.2±4.1，p = 0.030）的CEC频率显著更高，这与TAVI组中高危患者更高的欧洲心脏手术风险评估系统（Euroscore）、胸外科医师协会（STS）评分相对应。术后五天，与微创TAVI方法相比，侵入性更强的CAVR组的CEC频率显著更高（39.0±13.0对14.3±4.4，p < 0.001）。
新的流式细胞术方法可能是一种用于CEC计数的强大且可靠的方法。初步结果表明，CEC频率是评估术前风险、手术侵入性和临床结局的有效临床标志物。需要进一步研究来验证其实际临床实用性以及与传统标志物相比的潜在优势。
