Intracoronary Injection of Autologous CD34+ Cells Improves One-Year Left Ventricular Systolic Function in Patients with Diffuse Coronary Artery Disease and Preserved Cardiac Performance—A Randomized, Open-Label, Controlled Phase II Clinical Trial

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  Figure 1. The flow chart showing the enrollment, assignment, allocation, follow-up and analysis in this phase II clinical trial. CAD = coronary artery disease; IC = intracoronary artery; LVEF = left ventricular ejection fraction; F/U = follow.

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  Figure 2. Circulating EPC surface markers and soluble angiogenesis factors between group 1 and group 2 patients prior to and in group 1 after G-CSF treatment, and the time courses of EPC populations and SDF-1 α concentration in coronary sinus (CS) among group 1 patients. (A) Circulating number of CD34+ KDR+ CD45dim cells, * vs. other groups with different symbols (†, ‡), p < 0.0001. (B) Circulating number of CD34+CD133+CD45dim cells, * vs. other groups with different symbols (†, ‡), p < 0.0001. (C) Circulating number of CD31+ CD133+ CD45dim cells, * vs. other groups with different symbols (†, ‡), p < 0.0001. (D) Circulating number of CD34+ CD133+ KDR+ cells, * vs. other groups with different symbols (†, ‡), p < 0.0001. (E) Circulating number of CD133+ cells, * vs. †, p < 0.001. (F) Circulating number of CD34+ cells, * vs. †, p < 0.0001. Clt = control group (i.e., group 1); pre-G-CSF = indicated prior to G-CSF treatment in study group (SG) (i.e., group 1). (G) ELISA result of circulating hepatocyte growth factor (HGF), * vs. †, p < 0.0001. (H) ELISA result of circulating vascular endothelial growth factor (VEGF), * vs. †, p < 0.0001. SG (study group) (i.e., group 1), pre-G-CSF = indicated…

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  Figure 3. Illustrating the angiographic findings for identification of angiogenesis and Wimasis image analysis for assessment of angiogenesis. (A,D) Illustrating the coronary angiographic findings prior to (A), (C) and at 9th month (B), (D) after CD34+ therapy. As compared to control group (D), abundant angiogenesis/neovascularization were notably increased in study group (B) at 9th month after CD34+ therapy in both the same patients. (E,H) Illustrating regional microvasculature angiogenesis level in fixed territory assessment with Wimasis image software, between these two-time intervals in study group and control group. As compared to control group (H), plentiful angiogenesis/neovascularization (red color) were noted as having increased in study group (F) at 9th month after CD34+ therapy in both the same patients. (I) Wimasis assay for angiogenesis parameter of vessel density: (1) at baseline and (2) at 9th month, all p > 0.5. (J) Wimasis assay for angiogenesis parameter of total vessel network length: (1) at baseline, p > 0.5; (2) at 9th month, * vs. †, p < 0.05. (K) Wimasis assay for angiogenesis parameter of total branch point: (1) at baseline, p > 0.5; (2) at 9th month, * vs. †, p <…

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  Figure 4. Individual and mean changes of LVEF between baseline and 12 months, and serial changes of LVEF on 3-D echocardiographic findings during 1-year follow-up. (A) Illustrating the time courses of 3-D transthoracic echocardiographic findings of left ventricular ejection fraction (LVEF) among group 1 and group 2 patients. The results demonstrated that the LVEF was notably stepwise increased during the follow-up time points at 0, 1, 3, 6 and 12 months in group 1 patients. However, this parameter showed initially downwards, followed by stationary no more change in group 2 patients. (B) Compared with baseline (i.e., 0 month), the LVEF was significantly increased among group 1 patients, p < 0.001, but it showed no difference among group 2 patients, p = 0.29. (C) Illustrating the net LVEF improvements at different time points were significantly higher in group 1 than in group 2. Increases in the net change of LVEF improvement in group 1 as compared to group 2 at the time intervals between baseline vs. 1 month: +1.58 ± 4.88 vs. −1.56 ± 3.13, p = 0.023; 3 month: +2.16 ± 8.77 vs. −1.50 ± 2.16, p = 0.035; 6 month: +2.86 ± 2.26 vs. −1.38 ± 3.49, p < 0.001; 12 month: +4.63 ± 2.51 vs. −0.94 ± 4.57,…

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  Figure 5. Morphological feature of EPCs and Matrigel assay for assessment of angiogenesis. (A–C) Illustrating the microscopic finding (200x) of morphological feature of 21-day culturing endothelial progenitor cells (EPCs), i.e., cobblestone-like morphology of typical endothelial cells. Scale bars in the right lower corner represent 50 µm. (D–G) Illustrating the pictures of Matrigel assay. Red arrows indicated tubular formation; yellow arrows indicated cluster formation; green arrows indicated network formation. (H) Analytical result for number of tubule formation, * vs. other groups with different symbols (†, ‡), p < 0.0001. (I) Analytical result of total tubular length, * vs. other groups with different symbols (†, ‡), p < 0.0001. (J) Analytical result of mean tubular length, * vs. other groups with different symbols (†, ‡), p < 0.0001. (K) Analytical result of number of cluster formation, * vs. other groups with different symbols (†, ‡), p < 0.0001. (L) Analytical result of number of network formation, * vs. other groups with different symbols (†, ‡), p < 0.0001. All statistical analyses were performed by one-way ANOVA, followed by Bonferroni multiple comparison post hoc test (n = 30 for…