Injection Of Bone Marrow Mononuclear Cells Research Articles

Efficiency of Intramyocardial Injections of Autologous Bone Marrow Mononuclear Stem Cells in Patients with Ischemic Heart Failure: Long-Term Results

Introduction: Intramyocardial injection of autologous bone marrow mononuclear cells (BMMC) is believed to be a promising method for the treatment of patients with chronic ischemic heart disease. Hypothesis: To evaluate long-term results of intramyocardial bone marrow cell injection in patients with severe ischemic heart failure. Methods: Ninety nine ischemic heart failure patients (age 58 ± 5) with left ventricle ejection fraction (LVEF) ≤35% and III-IV NYHA and CCS functional classes without possibility for conventional revascularization were randomized into two groups: 1 group (n=49) - received intramyocardial BMMC injection and patients of the second group (n=50) were treated with standard medical therapy for HF. The NOGA system was used to administer 4.9 x107 BMMC into the infarction border zone. We analyzed all clinical data at 6 and 12 month of the follow-up period. Results: None of the patients developed periprocedural complications following BMMC injections. In the first group, the injections led to improvement of CCS class (3.5 ± 0.5 vs 2.1 ± 0.6 after 6 months and 1.9 ± 0.6 after 12 months (p<0.05 vs. baseline) and NYHA functional class (3.4 ± 0.1 vs 2.3 ± 0.2 after 6 months and 2.0 ± 0.2 after 12 months; p<0.05). The same characteristics in patients of the 2 group therapy remained unaltered (p = NS). LVEF increased in group 1 (34 ± 2% vs 42 ± 3%; p<0.05) while it tended to decrease in the control group (34 ± 2% vs 31 ± 3%; p = NS). Summed rest score improved in the first group after 6 months (14.7 ± 5.4 vs 10.7 ± 4.9; p<0.05). The improvement of stress score was more noticeable (18.2 ± 5.4 vs 9.4 ± 4.2; p<0.05). Neither stress nor rest score changed in patients of the second group. At the moment after 12 month follow up period 5 patients (10.2%) from group 1 died compared with 14 patients (28%) from group 2 (log rank test, p=0.008). Conclusions: Intramyocardial injection of autologous bone marrow mononuclear cells in ischemic heart failure patients is a safe procedure that improves clinical symptoms and has beneficial effecton LV function.

Long-term myocardial functional improvement after autologous bone marrow mononuclear cells transplantation in patients with ST-segment elevation myocardial infarction: 4 years follow-up

AimsTo evaluate the safety profile and efficacy of bone marrow mononuclear cells (BMMNC) transplantation for ST-segment elevation myocardial infarction (STEMI) by assessing patients and their left ventricular function at up to 4 years follow-up.Methods and resultsEighty-six patients with STEMI who had successfully undergone percutaneous coronary intervention (PCI) were randomized to receive intracoronary injection of BMMNC (n = 41) or saline (n = 45). Left ventricular ejection fraction, as evaluated by UCG, was markedly improved at 6 months (0.484 ± 0.5 vs. 0.457 ± 0.6, P = 0.001), 1 year (0.482 ± 0.7 vs. 0.446 ± 0.6, P < 0.001), and 4 years (0.505 ± 0.8 vs. 0.464 ± 0.8, P < 0.001) after BMMNC transplant when compared with control group. However, the current cell therapy did not improve the myocardial viability of the infarcted area as assessed by single-photon emission computed tomography analysis at 4 years post-transplant (0.263 ± 0.007 in BMMNC group vs. 0.281 ± 0.008 in control group, P = 0.10). During the follow-up period, one control group case (2.2%) of in-stent restenosis was confirmed by coronary angiography and underwent repeat PCI. Also during follow-up, one death (2.2%) occurred in the control group, and one patient (2.4%) in the BMMNC group had transient acute heart failure.ConclusionThis study indicates that intracoronary delivery of autologous BMMNC is safe and feasible for STEMI patients who have undergone PCI, and can lead to long-term improvement in myocardial function.

Donor cell-type specific paracrine effects of cell transplantation for post-infarction heart failure

Cell transplantation is an emerging therapy for treating post-infarction heart failure. Although the paracrine effect has been proposed to be an important mechanism for the therapeutic benefits, details remain largely unknown. This study compared various aspects of the paracrine effect after transplantation of either bone marrow mononuclear cells (BMC) or skeletal myoblasts (SMB) into the post-infarction chronically failing heart. Three weeks after left coronary artery ligation, adult rats received intramyocardial injection of either BMC, SMB or PBS only. Echocardiography demonstrated that injection of either cell type improved cardiac function compared to PBS injection. Interestingly, BMC injection markedly improved neovascularization in the border areas surrounding infarcts, while SMB injection decreased fibrosis in both the border and remote areas. Injection of either cell type similarly reduced hypertrophy of cardiomyocytes as assessed by cell-size planimetry using isolated cardiomyocytes. Quantitative RT-PCR revealed that, among 15 candidate mediators of paracrine effects studied, Fgf2 and Hgf were upregulated only after BMC injection, while Mmp2 and Timp4 were modulated after SMB injection. Additional investigations of signalling pathways relevant to heart failure by western blotting showed that p38 and STAT3 were temporarily activated after BMC injection, in contrast, ERK1/2 and JNK were activated after SMB injection. There was no difference in activation of Akt, PKD or Smad3 among groups. These data suggest that paracrine effects observed after cell transplantation in post-infarction heart failure were noticeably different between cell types in terms of mediators, signal transductions and consequent effects.

Abstract 2874: Bone Marrow-Derived Mononuclear Cell Transfer Is Associated with Superior Cardiac Recovery and Neovascularization as Compared to Nonmodified Mesenchymal Stem Cell Therapy in Chronic Myocardial Infarction

Background: Cell type with optimal efficacy for cardiac repair remains under discussion. We conducted a randomized, blind and placebo-controlled study with head-to-head comparison of autologous bone marrow mononuclear cells (BMNC) and autologous non-modified mesenchymal stem cells (BMSC) therapy in a large animal model of chronic MI. Methods: Eleven weeks after coronary ligation, 24 dogs received intramyocardial injections of placebo (n = 8), BMNC (227.10 6 ± 32.10 6 cells, n = 8) or nonmodified-MSC (232.10 6 ± 40.10 6 cells, n = 8). Cardiac performance and remodelling were assessed up to 16 weeks follow-up (FU). Results: (Table ) BMNC injection induced a decrease in the WMSI at 8 and 16 weeks as compared to baseline (BL). In contrast, injection of the BMSC was associated with a moderate decrease at 16 weeks vs BL. Furthermore, LV end-systolic elastance (Ees) increased only after injection of BMNC and remained unchanged after BMSCs injection. Conclusions: In the canine model of chronic MI, myocardial BMNC transfer is superior to non-modified BMSC to reduce infarct size, improve regional systolic function and cardiac contractility. Functional improvement is associated with favourable angiogenic environment and neovascularization.

Apoptosis and Effects of Intracavernous Bone Marrow Cell Injection in a Rat Model of Postprostatectomy Erectile Dysfunction

To investigate the pathophysiology of postprostatectomy erectile dysfunction (pPED) in a rat model of bilateral cavernous nerve ablation (BCNA) and to assess the effects of local bone marrow mononuclear cell (BMMNC) injection on erectile dysfunction (ED) and cavernosal cellular abnormalities caused by BCNA. This was an experimental study in Fisher rats with BCNA. Intervention included BNCA, electrical stimulation of the pelvic ganglion, and local BMMNC injection. Erectile responses to electric pelvic ganglion stimulation were studied. Cavernous tissue was examined to determine the cell types undergoing apoptosis and to detect changes in protein and gene expression of neuronal nitric oxide synthase (nNOS) and endothelial nitric oxide synthase (eNOS) using real-time quantitative polymerase chain reaction (RTQ-PCR) and Western blotting. The effects of local BMMNC injection on these parameters were studied. Diffuse apoptosis was noted in the connective tissue mesenchymal cells and vascular smooth muscle and endothelial cells. Compared with sham-operated controls, nNOS and eNOS levels were decreased after 3 wk and were normal (eNOS) or increased (nNOS) after 5 wk, suggesting spontaneous nerve regeneration. Despite nNOS recovery, erectile responses to electrical stimulation remained impaired after 5 wk, when mesenchymal cell apoptosis was the main persistent biologic abnormality. BMMNC injection decreased apoptotic cell numbers, accelerated the normalisation of nNOS and eNOS, and partially restored erectile responses at week 5. Massive cell apoptosis may play a key role in the pathophysiology of pPED. In this animal model, apoptosis persisted despite spontaneous nerve regeneration, suggesting that the course of BCNA-induced cell dysfunction was independent of reinnervation. BMMNC improved erectile function by inhibiting apoptosis and may hold promise for repairing penile cell damage caused by radical prostatectomy (RP).

Interleukin-10 From Transplanted Bone Marrow Mononuclear Cells Contributes to Cardiac Protection After Myocardial Infarction

Bone marrow mononuclear cells (BM-MNCs) have successfully been used as a therapy for the improvement of left ventricular (LV) function after myocardial infarction (MI). It has been suggested that paracrine factors from BM-MNCs may be a key mechanism mediating cardiac protection. We previously performed microarray analysis and found that the pleiotropic cytokine interleukin (IL)-10 was highly upregulated in human progenitor cells in comparison with adult endothelial cells and CD14+ cells. Moreover, BM-MNCs secrete significant amounts of IL-10, and IL-10 could be detected from progenitor cells transplanted in infarcted mouse hearts. Specifically, intramyocardial injection of wild-type BM-MNCs led to a significant decrease in LV end-diastolic pressure (LVEDP) and LV end-systolic volume (LVESV) compared to hearts injected with either diluent or IL-10 knock-out BM-MNCs. Furthermore, intramyocardial injection of wild-type BM-MNCs led to a significant increase in stroke volume (SV) and rate of the development of pressure over time (+dP/dt) compared to hearts injected with either diluent or IL-10 knock-out BM-MNCs. The IL-10-dependent improvement provided by transplanted cells was not caused by reduced infarct size, neutrophil infiltration, or capillary density, but rather was associated with decreased T lymphocyte accumulation, reactive hypertrophy, and myocardial collagen deposition. These results suggest that BM-MNCs mediate cardiac protection after myocardial infarction and this is, at least in part, dependent on IL-10.

Diabetic and Nondiabetic Patients Respond Differently to Transendocardial Injection of Bone Marrow Mononuclear Cells: Findings From Prospective Clinical Trials in “No-Option” Patients

Whether stem cell treatment has the same effect in diabetics and nondiabetics is unknown. To compare outcomes in these two groups, we analyzed data from 26 consecutive patients with chronic ischemic cardiomyopathy who were taking part in two clinical trials. Revascularization was not an option for these patients and they were treated with bone marrow mononuclear cells (BMMNCs). Patients underwent NOGA electromechanical mapping to identify viable myocardium (i.e., with a unipolar voltage > or = 6.9 mV), after which they received a mean of 28.5+/-4.7 x 10(6) BMMNCs. Patients were followed up at 6 months. In nondiabetics, there was a significant decrease in endsystolic volume between baseline and 6-month follow-up. In addition, New York Heart Association (NYHA) functional class decreased significantly (P=.04) from 3.0 (1.75-3.0) to 1.0 (1.0-2.0), the Canadian Cardiovascular Society angina score (CCSAS) improved significantly (P=.04) from 3.0 (2.0-4.0) to 1.0 (1.0-1.5), and oxygen uptake increased significantly (P=.04) from 16.4 (13.1-21.5) to 24.5 (17.3-29.2) ml/kg/min. These changes were not observed in diabetic patients. This is the first clinical study to show that BMMNC injection could have a smaller effect in diabetics.

Effect of Adult Bone Marrow Stem Cells on Myocardial Regeneration in Doxorubicin-Induced Mouse Cardiomyopathy

Background and Objectives: Bone marrow cells have been shown to differentiate into various cell lineages, including cardiomyocytes, in recent studies. This study evaluates the hypothesis that intravenous injection of bone marrow mononuclear cells (BMNCs) into rats with doxorubicin-induced cardiomyopathy can induce myocardial regeneration and improve myocardial contractility. Materials and Methods: Adult male Sprague-Dawley rats were induced to develop cardiomyopathy by treatment with doxorubicin (2.5 mg/kg, 6 times, 2-week period). Stem cell enriched BMNCs were injected into the tail vein of the rats after cessation of the doxorubicin injections. One week after the injection of PKH-67-labeled BMNCs, the localization of transplanted cells was evaluated. Immunohistochemical studies and Western blots were performed two weeks after BMNCs injection. Results: Cell-treated animals showed significant improvement in left ventricular fractional shortening as compared to untreated (control) rats (cell treated group vs. control group 47.2±4.9% vs. 34.4±3.6%, p<0.01). Histological analyses showed that in the cell-treated animals there was an increase in ventricular interstitial collagen deposition and the cell-treated animals had an improved number of capillary endothelial cells as compared with the control rats. PKH-67labeled BMNCs and cell proliferation by BrdU was noted in the cell-treated hearts. Cardiac CXCR4 protein expression increased at day 7 and 14 in the cell-treated rats, but only at day 14 in the control animals. Conclusion: These results suggest that intravenous injection of BMNCs effectively induce engraftment of BMNCs into the myocardium and attenuation of fibrosis. Intravenous injection of BMNCs also improved myocardial contractility in doxorubicininduced cardiomyopathy. (Korean Circ J 2008;38:110-118)

Autologous bone marrow-derived progenitor cell myocardial delivery for recent myocardial infarction patients following early angioplasty: results from a pilot study

Cellular cardiomyoplasty is a potential therapeutic approach to preventing left ventricular remodeling after myocardial infarction and has shown encouraging results such as induction of neoangiogenesis and functional improvement of diseased hearts. We report the results of a pilot study on progenitor cells in five patients with acute myocardial infarction (AMI). Patients with single-vessel disease who had their first episode of myocardial infarction and underwent angioplasty after 48 h (an average of 17 days following myocardial infarction) were included in the study. Mononuclear cells (MNCs) (1x10(7)) were isolated by Ficoll Hypaque method from 60 ml of bone marrow (BM) obtained from the iliac crest of 5 patients (aspiration was performed under local anesthesia). The mean CD34 count was 1-4%. After confirming the patency of the affected vessel postangioplasty, cellular concentrate was injected into the affected artery in 3-ml boluses (three to four injections), with intermittent occlusion. The mean age of all five male patients was 48.6+13.7 years. At 1 year, five patients were asymptomatic, and one had Class II dyspnea on exertion. The results of an echocardiogram performed at 6 months showed an improvement in ejection fraction (EF) from 35.3% to 43.13% and in fractional shortening from 24.75% to 28.33%. End-systolic volume decreased from 115.5 to 92.3 ml, end-diastolic volume decreased from 177.5 to 170 ml, and end-systolic dimensions also decreased from 4.26 to 4 mm, demonstrating positive left ventricular remodeling. Repeat echocardiogram at 1 year showed persistent improvement in EF. No adverse events were noted either before or after the procedure. The injection of autologous BM MNCs is a safe and efficacious therapy following early revascularization in AMI patients.

Safety of intramyocardial injection of autologous bone marrow cells to treat myocardial ischemia in pigs

The purpose of this study is to determine the potential adverse consequences of intracardiac injections of bone marrow mononuclear cells (BMCs) to facilitate the revascularization of ischemic myocardium. Bone marrow mononuclear cells are used to treat heart failure, though there are few studies that evaluated the safety of BMC transplantation for chronic myocardial ischemia. The pigs received coronary ameroid constrictors to induce chronic myocardial ischemia and left ventricular dysfunction. At 4 weeks, autologous BMCs were injected intramyocardially by Boston Scientific Stiletto catheter with low-dose (10(7) cells) or high-dose BMC (10(8)). Control animals received saline. Blood samples were collected for hematological and chemical indices, including cardiac enzyme levels at regular time intervals postinfarction. At 7 weeks, animals underwent electrophysiological study to evaluate the arrhythmic potential of transplanted BMC, followed by necropsy and histopathology. No mortalities were associated with intramyocardial delivery of BMC or saline. At Day 0, the total creatine phosphokinase (CPK) was in the normal range in all groups. All groups had significant elevations in CPK after ameroid placement, with no significant differences between groups. At 7 weeks, CPK in all groups had returned to pretreatment levels. Electrophysiological assessment revealed that one control animal had an inducible arrhythmia. No arrhythmias were induced in low- or high-dose BMC-treated pigs. There were no histopathological changes associated with BMC injection. This study showed, in a clinically relevant large-animal model, that catheter-based intramyocardial injection of autologous BMC into ischemic myocardium is safe.

Impaired CXCR4 Signaling Contributes to the Reduced Neovascularization Capacity of Endothelial Progenitor Cells From Patients With Coronary Artery Disease

Transplantation of bone marrow cells as well as circulating endothelial progenitor cells (EPC) enhances neovascularization after ischemia. The chemokine receptor CXCR4 is essential for migration and homing of hematopoietic stem cells. Therefore, we investigated the role of CXCR4 and its downstream signaling cascade for the angiogenic capacity of cultured human EPC. Ex vivo, differentiated EPC derived from peripheral blood abundantly expressed CXCR4. Incubation of EPC from healthy volunteers with neutralizing antibodies against CXCR4 profoundly inhibited vascular endothelial growth factor- and stromal-derived factor-1-induced migration as well as EPC-induced angiogenesis in an ex vivo assay. Preincubation of transplanted EPC with CXCR4 antibody reduced EPC incorporation and impaired blood-flow recovery in ischemic hindlimbs of nude mice (57+/-4% of normal perfusion versus untreated EPC: 80+/-11%, P<0.001). Bone marrow mononuclear cells (BM-MNC) or EPC of heterozygous CXCR4(+/-) mice displayed reduced CXCR4 expression and disclosed impaired in vivo capacity to enhance recovery of ischemic blood flow in nude mice (blood flow 27+/-11% versus 66+/-25% using wild-type cells, P<0.01). Importantly, impaired blood flow in ischemic CXCR4(+/-) mice was rescued by injection of wild-type BM-MNC. Next, we investigated the role of CXCR4 for functional capacities of EPC from patients with coronary artery disease (CAD). Surface expression of CXCR4 was similar in EPC from patients with CAD compared with healthy controls. However, basal Janus kinase (JAK)-2 phosphorylation was significantly reduced and less responsive to stromal-derived factor-1 in EPC from patients with CAD compared with healthy volunteers, indicating that CXCR4-mediated JAK-2 signaling is dysregulated in EPC from patients with CAD. The CXCR4 receptor signaling profoundly modulates the angiogenic activity and homing capacity of cultured human EPC. Disturbance of CXCR4 signaling, as demonstrated by reduced JAK-2 phosphorylation, may contribute to functional impairment of EPC from patients with CAD. Stimulating CXCR4 signaling might improve functional properties of EPC and may rescue impaired neovascularization capacity of EPC derived from patients with CAD.

Melhora sintomática e da capacidade de exercício após o transplante autólogo, transendocárdico, de células mononucleares da medula óssea em pacientes com cardiopatia isquêmica grave, sustentada até o sexto mês de evolução

This study aimed at assessing the effects of autologous transendocardial transplantation of bone marrow mononuclear cells (ATTBMMC) on symptoms, exercise capacity, myocardial perfusion and contractility in patients with severe ischemic heart disease during a 6-month follow-up period. This prospective study comprised 21 patients as follows: the first 14 patients forming the treated group, and the last 7 patients forming the control group. Initially, all patients underwent clinical and laboratory assessment, treadmill testing, echocardiography, myocardial scintigraphy, and 24-hour Holter. The bone marrow mononuclear cells (BMMC) were isolated, washed, and diluted in 0.9% saline solution for transendocardial injection in areas of viable myocardium in the treated group, (15 0.2-mL injections). All patients were reassessed in the end of 2 and 6 months of follow-up. The demographic data and other characteristics did not significantly differ between the groups in the initial evaluation. No major adverse events related to the ATTBMMC were observed. In the end of 6 months, a reduction in the ischemic area was observed on nuclear perfusion imaging (P=0.05), as was a significant improvement in symptoms, functional capacity, and left ventricular overall function. This study showed that transendocardial injections of BMMC are safe in human beings with ischemic heart disease associated with severe ventricular dysfunction. The effects observed in the short run were maintained up to the sixth month of follow-up.

Implantation of bone marrow mononuclear cells using injectable fibrin matrix enhances neovascularization in infarcted myocardium

Neovascularization may improve cardiac function and prevent further scar tissue formation in infarcted myocardium. A number of studies have demonstrated that bone marrow-derived cells have the potential to induce neovascularization in ischemic tissues. In this study, we hypothesized that implantation of bone marrow mononuclear cells (BMMNCs) using injectable fibrin matrix further enhances neovascularization in infarcted myocardium compared to BMMNC implantation without matrix. To test this hypothesis, infarction was induced in rat myocardium by cryoinjury. Three weeks later, rat BMMNCs were mixed with fibrin matrix and injected into the infarcted myocardium. Injection of either BMMNCs or medium alone into infarcted myocardium served as controls. Eight weeks after the treatments, histological analyses indicated that implantation of BMMNCs using fibrin matrix resulted in more extensive tissue regeneration in the infarcted myocardium compared to BMMNC implantation without matrix. Examination with fluorescence microscopy revealed that cells labeled with a fluorescent dye prior to implantation survived in the infarcted myocardium at 8 weeks of implantation. Importantly, implantation of BMMNCs using fibrin matrix resulted in much more extensive neovascularization in infarcted myocardium than BMMNC implantation without matrix. The microvessel density in infarcted myocardium was significantly higher ( p<0.05) when BMMNCs were implanted using fibrin matrix (350±22 microvessels/mm 2) compared to BMMNC implantation without matrix (262±13 microvessels/mm 2) and medium injection (76±9 microvessels/mm 2). In addition, average internal diameter of microvessels was significantly larger ( p<0.05) in BMMNC implantation with fibrin matrix group (14.6±1.2 μm) than BMMNC implantation without matrix group (10.2±0.7 μm) and medium injection group (7.3±0.5 μm). These results suggest that fibrin matrix could serve as a cell implantation matrix that enhances neovascularization efficacy for myocardial infarction treatment.