Bone Marrow Mononuclear Cell Transplantation Research Articles

Benefits of Autologous Bone Marrow Mononuclear Cell Transplantation in Chronic Ischemic Pontine Infarct

Introduction: Ischemic stroke is brain injury due to hypoperfusion leading to sudden neurological symptoms. Despite advances in medicine, prognosis is still grim especially with chronic residual neurodeficits. Autologous bone marrow mononuclear cells (BMMNCs) transplantation is safe and has shown positive outcomes in previous studies of ischemic stroke. Our aim was to study the effect of autologous BMMNCs transplantation in chronic ischemic stroke. Intervention and Results: We treated a case of pontine ischemic infarct three years later, with BMMNCs transplantation followed by standard rehabilitation. Seven months after transplantation he showed significant improvement in spasticity, motor function and voluntary control of upper and lower extremity, static and dynamic sitting and standing balance. Improvement was noted in the scales like Modified Ashworth scale (mAS), Motor Assessment scale (MAS), Disabilities of Shoulder, arm and hand questionnaire (DASH), Modified Rankin Scale (MRS), Berg Balance Scale (BBS) and Brunnstrom Voluntary Control Grading (BVCG). Functional Independence Measure score increased from 59 to 81. BMMNCs exert paracrine effects, neoangiogenesis and trophic effect which facilitate various reparative and regenerative processes in the penumbral area. Conclusion: In this case, the clinical changes exhibited by BMMNCs transplantation even at a chronic stage in stroke substantiate their therapeutic potential. Although a single case report, these findings provide foundation for further research. Larger randomized controlled trials are warranted to study the safety and efficacy.

Increasing Dose of Autologous Bone Marrow Mononuclear Cells Transplantation Is Related to Stroke Outcome: Results from a Pooled Analysis of Two Clinical Trials

Background and Purpose. BM-MNC transplantation improves recovery in experimental models of ischemic stroke. Clinical trials are ongoing to test efficacy in stroke patients. However, whether cell dose is related to outcomes is not known. Methods. We performed a pooling data analysis of two pilot clinical trials with autologous BM-MNCs transplantation in ischemic stroke patients. Cell dose and route were analyzed to evaluate their relation to good outcome (m-Rankin scale [mRS] score 0–2) at 6 months. Results. Twenty-two patients were included. A median of 153 × 106 (±121 × 106) BM-MNCs was injected. Intra-arterial route was used in 77.3% of cases. A higher number of cells injected were associated with better outcomes at 180 days (390 × 106 [320–422] BM-MNCs injected in those patients with mRS of 0–2 at 6 months versus 130 × 106 [89–210] in those patients with mRS 3–6, p = 0.015). In the intra-arterially treated patients, a strong correlation between dose of cells and disability was found (r = −0.63, p = 0.006). A cut point of 310 × 106 injected cells predicted good outcome with 80% sensitivity and 88.2% specificity. Conclusions. Similar to preclinical studies, a higher dose of autologous BM-MNC was related to better outcome in stroke patients, especially when more than 310 × 106 cells are injected. Further interventional studies are warranted to confirm these data.

Bone Marrow Mononuclear Cell Transplantation Restores Inflammatory Balance of Cytokines after ST Segment Elevation Myocardial Infarction

BackgroundAcute myocardial infarction (AMI) launches an inflammatory response and a repair process to compensate cardiac function. During this process, the balance between proinflammatory and anti-inflammatory cytokines is important for optimal cardiac repair. Stem cell transplantation after AMI improves tissue repair and increases the ventricular ejection fraction. Here, we studied in detail the acute effect of bone marrow mononuclear cell (BMMNC) transplantation on proinflammatory and anti-inflammatory cytokines in patients with ST segment elevation myocardial infarction (STEMI).MethodsPatients with STEMI treated with thrombolysis followed by percutaneous coronary intervention (PCI) were randomly assigned to receive either BMMNC or saline as an intracoronary injection. Cardiac function was evaluated by left ventricle angiogram during the PCI and again after 6 months. The concentrations of 27 cytokines were measured from plasma samples up to 4 days after the PCI and the intracoronary injection.ResultsTwenty-six patients (control group, n = 12; BMMNC group, n = 14) from the previously reported FINCELL study (n = 80) were included to this study. At day 2, the change in the proinflammatory cytokines correlated with the change in the anti-inflammatory cytokines in both groups (Kendall’s tau, control 0.6; BMMNC 0.7). At day 4, the correlation had completely disappeared in the control group but was preserved in the BMMNC group (Kendall’s tau, control 0.3; BMMNC 0.7).ConclusionsBMMNC transplantation is associated with preserved balance between pro- and anti-inflammatory cytokines after STEMI in PCI-treated patients. This may partly explain the favorable effect of stem cell transplantation after AMI.

Intramuscular injection of bone marrow mononuclear cells contributes to bone repair following midpalatal expansion in rats.

Healing from injury requires the activation and proliferation of stem cells for tissue repair. Previous studies have demonstrated that bone marrow is a central pool of stem cells. The present study aimed to investigate the route undertaken by bone marrow mononuclear cells (BMMCs) following BMMC transplantation by masseter injection in a rat model of midpalatal expansion. The rats were divided into five groups according to the types of midpalatal expansion, incision and BMMC transplantation. Samples of midpalatal bone from the rats in each group were used for histological and immunohistochemical assessments to track and evaluate the differential potentials of the transplanted BMMCs in the masseter muscle and midpalatal bone. Bromodeoxyuridine was used as a BMMC tracing label, and M-cadherin was used to detect muscle satellite cells. The BMMCs injected into the masseter were observed, not only in the masseter, but also in the blood vessels and oral mucosa, and enveloped the midpalatal bone. A number of the BMMCs transformed into osteoblasts at the boundary of the neuromuscular bundle, and were embedded in the newly formed bone during midpalatal bone regeneration. The results of the present study suggested that BMMCs entered the circulation and migrated from muscle to the bone tissue, where they were involved in bone repair. Therefore, BMMCs may prove useful in the treatment of various types of cancer.

Preconditioning with VEGF Enhances Angiogenic and Neuroprotective Effects of Bone Marrow Mononuclear Cell Transplantation in a Rat Model of Chronic Cerebral Hypoperfusion.

Bone marrow mononuclear cell (BMMNC) transplantation is a promising therapy for brain ischemia. However, BMMNCs are few in number, and a limited time window is available during which they can penetrate the blood-brain barrier (BBB) and migrate to the brain. We investigated whether vascular endothelial growth factor (VEGF) can facilitate BMMNC migration into the ischemic brain and enhance their therapeutic effect in a rat model of chronic cerebral hypoperfusion. First, we assessed the impact of VEGF on the BBB of rats that had undergone permanent bilateral occlusion of the common carotid arteries (2VO). Then, we transplanted BMMNCs into 2VO rats pretreated with intracerebroventricular VEGF or vehicle. We examined cognitive function with the Morris water maze test, BMMNC migration by immunofluorescence analysis, and cytokine levels in the peripheral blood by enzyme-linked immunosorbent assay (ELISA). Angiogenesis and neural degeneration were evaluated by staining tissue with Ki67/lectin or Fluoro-Jade B. We found that at a dose of 0.2μg/rat, VEGF significantly increased BBB permeability without causing brain edema in 2VO rats. VEGF + BMMNC-treated rats had more BMMNC migration in the ischemic brain, better learning and memory, greater proliferation of vessels, and fewer degenerating neurons than did BMMNC-treated rats. Pretreatment with VEGF receptor inhibitor SU5416 significantly decreased BMMNC migration and abolished the therapeutic effect of BMMNC transplantation. We conclude that preconditioning with an appropriate dose of VEGF can enhance the therapeutic efficacy of BMMNC transplantation in 2VO rats, possibly by facilitating BMMNC migration into the ischemic brain.

Autologous Bone Marrow Mononuclear Cell Transplantation Delays Progression of Carotid Atherosclerosis in Rabbits.

Bone marrow mononuclear cells (BMMNCs) can counteract oxidative stress and inhibit the inflammatory response in focal ischemic stroke models. However, the effect of BMMNC transplantation on carotid atherosclerosis needs to be determined. The carotid atherosclerotic plaque model was established in New Zealand White rabbits by balloon injury and 8weeks of high-fat diet. Rabbits were randomized to receive an intravenous injection of autologous bromodeoxyuridine (BrdU)-labeled BMMNCs or an equal volume of phosphate-buffered saline. Plaques were evaluated for expression of proinflammatory and anti-inflammatory cytokines, anti-oxidant proteins, and markers of cell death. BMMNCs migrated into atherosclerotic plaque on the first day after cell transplantation. BMMNC-treated rabbits had smaller plaques and more collagen deposition than did the vehicle-treated controls on day 28 (p < 0.05). BMMNC treatment significantly increased endothelial nitric oxide synthase and the anti-oxidant enzymes glutathione peroxidase and superoxide dismutase in plaques compared to vehicle treatment on day 7. BMMNC-treated rabbits also had lower levels of cleaved caspase-3 expression; lower levels of proinflammatory cytokines interleukin-1β, tumor necrosis factor alpha, and matrix metalloproteinase 9; and higher levels of insulin-like growth factor-1 and its receptor (p < 0.05). Autologous BMMNC transplantation can suppress the process of atherosclerotic plaque formation and is associated with enhanced anti-oxidative effect, reduced levels of inflammatory cytokines and cleaved caspase-3, and increased expression of insulin-like growth factor-1 and its receptor. BMMNC transplantation represents a novel approach for the treatment of carotid atherosclerosis.

Improved Quality of Life in A Case of Cerebral Palsy after Bone Marrow Mononuclear Cell Transplantation.

Cerebral palsy (CP) is a non progressive, demyelinating disorder that affects a child's development and posture and may be associated with sensation, cognition, communication and perception abnormalities. In CP, cerebral white matter is injured resulting in the loss of oligodendrocytes. This causes damage to the myelin and disruption of nerve conduction. Cell therapy is being explored as an alternate therapeutic strategy as there is no treatment currently available for CP. To study the benefits of this treatment we have administered autologous bone marrow mononuclear cells (BMMNCs) to a 12-year-old CP case. He was clinically re-evaluated after six months and found to demonstrate positive clinical and functional outcomes. His trunk strength, upper limb control, hand functions, walking stability, balance, posture and coordination improved. His ability to perform activities of daily living improved. On repeating the Functional Independence Measure (FIM), the score increased from 90 to 113. A repeat positron emission tomography-computed tomography (PET-CT) scan of the brain six months after intervention showed progression of the mean standard deviation values towards normalization which correlated to the functional changes. At one year, all clinical improvements have remained. This indicated that cell transplantation may improve quality of life and have a potential for treatment of CP.

Effectiveness of bone marrow mononuclear cells delivered through a graft vessel for patients with previous myocardial infarction and chronic heart failure: an echocardiographic study of left ventricular remodeling.

The graft of stem cells to treat ischemic cardiomyopathy is popular in many clinical trials. The aim of this study was to evaluate the effectiveness of isolated coronary artery bypass graft combined with bone marrow mononuclear cells (BMMNC) delivered through graft vessels to improve left ventricular remodeling of patients with previous myocardial infarction and chronic heart failure using echocardiography. Patients with previous myocardial infarction and chronic heart failure were randomly allocated to one of the two groups: CABG only (18 patients), or CABG with BMMNC transplantation (24 patients). Echocardiographic parameters were measured on B-mode imaging, 3D imaging and color flow imaging. RESULTS Post-operative LVEDD (end-diastolic dimension of left ventricle), LVESD (end-systolic dimension of left ventricle), LVEDV (end-diastolic volume of left ventricle), LVESV (end-systolic volume of left ventricle), LVEDVI (LVEDV indexed to body surface area), LVESVI (LVESV indexed to body surface area), LV-mass (mass of left ventricle) and LV-massI (LV-mass indexed to body surface area) were significantly improved compared with those obtained prior to operation in CABG+BMMNC group (al p<0.05). The same parameters were not significantly different pre- and postoperative in the CABG group (al p>0.05). Postoperative mitral regurgitation score was not significantly different from those prior to operation in both groups (al p>0.05). In Chi-square tests, LVEDD, LVESD, LVEDV, LVESV, LVEDVI, LVESVI, LV-mass, LV-massI were determinants of the left ventricular remodeling. The improvement of left ventricular remodeling in CABG+BMMNC group was better than in the CABG group and this improvement was verified by echocardiography.

Effects of Bone Marrow Mononuclear Cells Delivered through a Graft Vessel for Patients with Previous Myocardial Infarction and Chronic Heart Failure: An Echocardiographic Study of Left Atrium Function.

The graft of stem cells to treat ischemic cardiomyopathy is popular in many clinical trials. The aim of this study was to evaluate the effectiveness of isolated coronary artery bypass graft combined with bone marrow mononuclear cells (BMMNC) delivered through the graft vessels to improve left atrium (LA) function of patients with previous myocardial infarction and chronic heart failure using echocardiography. Forty-two patients with previous myocardial infarction and chronic heart failure were randomly allocated to one of the two groups: coronary artery bypass graft (CABG) only (18 in CABG group) or CABG with BMMNC transplantation (24 in CABG+BMMNC group). Echocardiographic parameters were measured on two-dimensional (2D) imaging, three-dimensional imaging, 2D strain, and strain rate imaging, eight parameters were measured totally. Postoperative LA global strain (LAGS) versus Preoperative LAGS were 24.875±1.588% versus 14.250±1.281% (P<0.05) in CABG+BMMMNC group and 17.556±1.834% versus 13.111±1.479% (P<0.05) in CABG group, LVEF was 49.083±1.914% versus 36.042±1.185% (P<0.05) in CABG+BMMMNC group and 41.389±2.210% versus 34.667±1.369% (P<0.05) in CABG group, and LA volume (LAV) was 15.651±0.631mL versus 23.894±0.831mL (P<0.05) in CABG+BMMMNC group and 21.429±0.707mL versus 22.379±0.959mL (P<0.05) in CABG group. LAGS, LVEF, and LAV were significantly improved postoperatively in both groups. The degree of the improvement was significantly different between the two groups with the CABG+BMMNC group improved more versus the group of CABG only (P<0.05). Postoperative LVEDV and LVESV were significantly improved compared with those obtained prior to operation in CABG+BMMNC group (P<0.05). The improvement of LA function in CABG+BMMNC group is better than CABG group. 2D strain imaging is a more sensitive tool to evaluate LA function.

Effect of heterologous bone marrow mononuclear cell transplantation on midpalatal expansion in rats.

The aim of this study was to explore whether bone marrow mononuclear cell (BMMC) transplantation is able to accelerate the bone remodeling induced by midpalatal expansion in rats. A total of 48 male Sprague-Dawley rats (mean weight, 208.36±7.32 g) were divided into control and midpalatal expansion with or without BMMC transplantation groups. Histological and morphological changes were observed in each group. The osteogenic activities and differential potentials of the transplanted BMMCs labeled with bromodeoxyuridine in the midpalatal bone tissue were assessed by osteocalcin expression. The receptor activator of nuclear factor κB ligand (RANKL)/osteoprotegerin (OPG) ratio was determined by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) to reflect the equilibrium between bone resorption and formation. The results demonstrated that the width of the maxillary dental arch increased distinctly within 2 weeks of midpalatal expansion with BMMC transplantation. The morphology of the midpalatal suture in this group changed significantly; the cartilage was completely replaced by fibrous-like tissue expressing osteocalcin. The palatal bone was reorganized from a cancellous form into a mature compact structure after an additional 2-week relapse period. Immunostaining results indicated that the heterologous transplanted BMMCs survived and differentiated into osteoblasts during the remodeling induced by midpalatal expansion. The RANKL/OPG expression ratio significantly decreased after 2 weeks of midpalatal expansion with BMMC transplantation due to the inhibition of RANKL expression. Heterologous BMMC transplantation appears to accelerate the midpalatal bone remodeling induced by expansion of the rats through increasing the number of osteoprogenitor cells and regulating the RANKL-OPG signaling pathway.

Transplantation of bone marrow mononuclear cells modulates hippocampal expression of growth factors in chronically epileptic animals.

In previous studies, transplantation of bone marrow mononuclear cells (BMMCs) in epileptic animals has been found to be neuroprotective. However, the mechanism by which the BMMCs act remains unclear. We hypothesize that BMMCs may provide neuroprotection to the epileptic brain through trophic support. To test our hypothesis, we studied the temporal expression of neurotrophins after BMMC transplantation in the epileptic rat hippocampus. Chronically epileptic rats were intravenously transplanted with 1 × 10(7) BMMCs isolated from GFP transgenic mice. Expression levels of BDNF, GDNF, NGF, VEGF, and TGF-β1, and their receptors, were evaluated by ELISA and/or qRT-PCR analysis. Our data revealed increased protein expression of BDNF, GDNF, NGF, and VEGF and reduced levels of TGF-β1 in the hippocampus of transplanted epileptic animals. Additionally, an increase in the mRNA expression of BDNF, GDNF, and VEGF, a reduction in TGF-β1, and a decrease in mRNA levels of the TrkA and TGFR-β1 receptors were also observed. The gain provided by transplanted BMMCs in the epileptic brain may be related to the ability of these cells in modulating the network of neurotrophins and angiogenic signals.

Bone marrow characteristics associated with changes in infarct size after STEMI: a biorepository evaluation from the CCTRN TIME trial.

Despite significant interest in bone marrow mononuclear cell (BMC) therapy for ischemic heart disease, current techniques have resulted in only modest benefits. However, selected patients have shown improvements after autologous BMC therapy, but the contributing factors are unclear. The purpose of this study was to identify BMC characteristics associated with a reduction in infarct size after ST-segment-elevation-myocardial infarction. This prospective study comprised patients consecutively enrolled in the CCTRN TIME (Cardiovascular Cell Therapy Research Network Timing in Myocardial Infarction Evaluation) trial who agreed to have their BMCs stored and analyzed at the CCTRN Biorepository. Change in infarct size between baseline (3 days after percutaneous coronary intervention) and 6-month follow-up was measured by cardiac MRI. Infarct-size measurements and BMC phenotype and function data were obtained for 101 patients (mean age, 56.5 years; mean screening ejection fraction, 37%; mean baseline cardiac MRI ejection fraction, 45%). At 6 months, 75 patients (74.3%) showed a reduction in infarct size (mean change, -21.0±17.6%). Multiple regression analysis indicated that infarct size reduction was greater in patients who had a larger percentage of CD31(+) BMCs (P=0.046) and in those with faster BMC growth rates in colony-forming unit Hill and endothelial-colony forming cell functional assays (P=0.033 and P=0.032, respectively). This study identified BMC characteristics associated with a better clinical outcome in patients with segment-elevation-myocardial infarction and highlighted the importance of endothelial precursor activity in regenerating infarcted myocardium. Furthermore, it suggests that for these patients with segment-elevation-myocardial infarction, myocardial repair was more dependent on baseline BMC characteristics than on whether the patient underwent intracoronary BMC transplantation. http://www.clinicaltrials.gov. Unique identifier: NCT00684021.

Autologous bone marrow transplantation in decompensated liver: Systematic review and meta-analysis.

To evaluate the efficacy of autologous bone marrow mononuclear cell transplantation in decompensated liver disease. Medline, EMBASE, PubMed, Science Direct, and the Cochrane Library were searched for relevant studies. Retrospective case-control studies were included along with randomized clinical trials. Meta-analysis was performed in line with recommendations from the Cochrane Collaboration software review manager. Heterogeneity was assessed using a random-effects model. Four randomized controlled trials and four retrospective studies were included. Cell transplantation increased serum albumin level by 1.96 g/L (95%CI: 0.74-3.17; P = 0.002], 2.55 g/L (95%CI: 0.32-4.79; P = 0.03), and 3.65 g/L (95%CI: 0.76-6.54; P = 0.01) after 1, 3, and 6 mo, respectively. Patients who had undergone cell transplantation also had a lower level of total bilirubin [mean difference (MD): -1.37 mg/dL; 95%CI: -2.68-(-0.06); P = 0.04] after 6 mo. This decreased after 1 year when compared to standard treatment (MD: -1.26; 95%CI: -2.48-(-0.03); P = 0.04]. A temporary decrease in alanine transaminase and aspartate transaminase were significant in the cell transplantation group. However, after 6 mo treatment, patients who had undergone cell transplantation had a slightly longer prothrombin time (MD: 5.66 s, 95%CI: 0.04-11.28; P = 0.05). Changes in the model for end-stage liver disease score and Child-Pugh score were not statistically significant. Autologous bone marrow transplantation showed some benefits in patients with decompensated liver disease. However, further studies are still needed to verify its role in clinical treatment for end-stage liver disease.

Effect on the expression of nestin and synapsin I after spinal cord injury in rats

Objective To investigate the effects and significance of site transplantation of allogenic bone marrow mononuclear cells （BMMNCs） on the expression of nestin and synapsin I in the local mieroenvironment of transected injury zone in spinal cord. Methods From a pool of 120 clean grade Wistar rats （ Irrespective of gender） , 87 rats were randomly （ random number） selected and subdivided into three groups. Sham operation group （n = 29 ） had lamineetomy done without inducing spinal cord injury. The model control group （n = 29 ） had laminectomy done followed by induction of complete spinal cord transeeted injury. Cell transplantation group （n = 29） had BMMNCs transplantation initiated after induction of complete spinal cord transected injury. Nestin and Synapsin expressions in all rats of each group were assessed on day 5, week 1, week 2 and week 4, respectively by using immunohistochemical and RT-PCRtechniques. Spread of Nestin positive and Synapsin I positive products along the spinal cord were assessed by use of MIAS-2000 image processing system analysis. Optical density of these products was then measured. Statistical data analysis was done using SPSS17. 0. Uniform variance analysis was done for inter group comparison using completely random design data, while paired sample comparison done with the Q test. For non-uniform variance, multiple samples were compared using design rank sum test with Kruskal-Wallis method, whereas paired samples rank sum test with Nemenyi method. P value of P 〈 0. 05 was regarded statistically significant, and P 〈 0.01 regarded highly statistically significant. Results Immunohistochemistry, RT-PCR and image processing system analysis showed model control group had increased nestin expression within the damaged zone 5 days after injury, reaching peak level at 2 weeks then significantly down regulated 4 weeks later. In cell transplantation group, nestin expression around spinal cord injury zone increased 5 days after injury and reached the peak level at 1 week, then subsequently significantly down regulated 4 weeks later. At each interval, there were significantly statistical differences between sham surgery group and model control group and cell transplantation group （P 〈 0.05 ）. Synapsin I expression appeared within the damaged area 5 days after injury, but at each interval, the expression of synapsin I in the model control group and cell transplantation group was lower than that in sham operation group. However, synapsin I in cell transplantation group was higher than that in model control group （ P 〈 0. 01）. Conclusions Allogenic bone marrow mononuclear cells transplantation may alter the local microenvironment and up-regulate the expression of nestin and synapsin I in the spinal cord injury model. Key words: Bone marrow mononuclear cells ; Spinal cord injury ; Cell transplantation; Nestin; Synaptophysin I; Spinal cord injury; Immunohistochemistry; RT-PCR

Effects of Bone Marrow Mononuclear Cells Delivered through a Graft Vessel for Patients with Previous Myocardial Infarction and Chronic Heart Failure: An Echocardiographic Study of Left Ventricular Function.

The graft of stem cells to treat ischemic cardiomyopathy is popular in many clinical trials. The aim of this study was to evaluate the effectiveness of isolated coronary artery bypass graft combined with bone marrow mononuclear cells (BMMNC) delivered through graft vessels to improve left ventricular function of patients with previous myocardial infarction and chronic heart failure using echocardiography. Forty-two patients with previous myocardial infarction and chronic heart failure were randomly allocated to one of the two groups: CABG only (18 in CABG group), or CABG with BMMNC transplantation (24 in CABG+BMMNC group). Echocardiographic parameters of systolic function were measured on B-mode imaging, tissue Doppler imaging (TDI), two-dimensional (2D) strain imaging, and 8 parameters were measured totally. Echocardiographic parameters of diastolic function were measured on pulsed-wave Doppler imaging, TDI, and 2D strain rate imaging; 17 parameters were measured totally. Postoperative left ventricular ejection fraction (LVEF) versus preoperative LVEFwere 49.083±1.914% versus 36.042±1.185% (P<0.05) in CABG+BMMMNC group and 41.389±2.210% versus 34.667±1.369% (P<0.05) in CABG group, global longitudinal strain were-12.542±0.512% versus -7.083±0.583% (P<0.05) in CABG+BMMMNC group and -9.278±0.591% versus -7.000±0.673% (P<0.05) in CABG group, mLsr1 were -0.108±0.018/sec versus -0.039±0.017/sec (P<0.05) in CABG+BMMMNC group and -0.048±0.021/sec versus0.004±0.020/sec (P<0.05) in CABG group, mLsr2 were -0.055±0.013/sec versus-0.009±0.015/sec (P<0.05) in CABG+BMMMNC group and 0.004±0.015/sec versus0.024±0.017/sec (P<0.05) in CABG group, and Aa1 were 7.303±0.479cm/sec versus 5.131±0.381cm/sec (P<0.05) in CABG+BMMMNC group and 7.908±0.553cm/sec versus 6.764±0.440cm/sec (P<0.05) in CABG group. Parameters above were significantly improved postoperatively in both groups. The degree of the improvement was significantly different between the two groups with the CABG+BMMNC group improved more versus the group of CABG only (P<0.05). The improvement of left ventricular function in CABG+BMMNC group is better than CABG group. 2D strain and strain rate imaging is a more sensitive tool to evaluate left ventricular function.

The effect of transplanted bone marrow mononuclear cells under different transplant paths on neovascularization in ischemic cerebral infarction rats

Objective To investigate the effect of treatment with transplanted bone marrow mononuclear cells under different transplant paths on neovascularization in ischemic cerebral infarction rats. Methods The MCAO models were established by intraluminal vascular suture method. All the successful models were randomly divided into control group, intravenous transplantation group and subarachnoid transplantation group according to the random number table. The Extraction, isolation and culture of bone marrow mononuclear cells were extracted, isolated and cultured. The models in the control group were injected 200 μl PBS through the femoral vein. The models in the intravenous transplantation group were injected 200 μl containing 2×107 bone marrow mononuclear cells through the femoral vein. The models in the subarachnoid transplantation group were injected 200 μl containing 2×107 bone marrow mononuclear cells through the subarachnoid. The VEGF levels in cerebrospinal fluid of rats were detected using quantitative ELISA method. The vascular regenerations in the infarct area were observed by using quantitative immunofluorescence. Results After transplantation of bone marrow mononuclear cells 7 d, 14 d and 28 d, the VEGF levels of rat cerebrospinal fluid in subarachnoid transplantation group were (207.4±8.9)pg/ml, (171.2±10.3)pg/ml and (143.8±13.8)pg/ml, respectively, which were all higher than the VEGF levels in intravenous transplantation group, the differences were all statistically significant (P<0.05). After transplantation 14 d and 28 d, the BrdU-positive cells in subarachnoid transplantation group were (2043.8±514.2) and (1834.8±307.4), respectively, which were higher than the intravenous transplantation group, the differences were all statistically significant (P<0.05). The microvessel counts in subarachnoid transplantation group were (384.6±45.1) and (514.8±51.3), respectively, which were higher than that in the intravenous transplantation group, and the differences were all statistically significant (P<0.05). Conclusions Bone marrow mononuclear cells transplantation can increase VEGF secretion, and promote angiogenesis infarct. The effect of subarachnoid transplantation is better. Key words: Cerebral infarction; Bone marrow mononuclear cells; Transplantation; Angiogenesis

Sustained effect of bone marrow mononuclear cell therapy in axonal regeneration in a model of optic nerve crush

In adult mammals, the regeneration of the optic nerve is very limited and at the moment there are several groups trying different approaches to increase retinal ganglion cell (RGC) survival and axonal outgrowth. One promising approach is cell therapy. In previous work, we performed intravitreal transplantation of bone-marrow mononuclear cells (BMMCs) after optic nerve crush in adult rats and we demonstrated an increase in RGC survival and axon outgrowth 14 days after injury. In the present work, we investigated if these results could be sustained for a longer period of time. Optic nerve crush was performed in Lister-hooded adult rats and BMMC or saline injections were performed shortly after injury. Neuronal survival and regeneration were evaluated in rats׳ retina and optic nerve after 28 days. We demonstrated an increase of 5.2 fold in the axon outgrowth 28 days after lesion, but the BMMCs had no effect on RGC survival. In an attempt to prolong RGC survival, we established a new protocol with two BMMC injections, the second one 7 days after the injury. Untreated animals received two injections of saline. We observed that although the axonal outgrowth was still increased after the second BMMC injection, the RGC survival was not significantly different from untreated animals. These results demonstrate that BMMCs transplantation promotes neuroregeneration at least until 28 days after injury. However, the effects on RGC survival previously observed by us at 14 days were not sustained at 28 days and could not be prolonged with a second dose of BMMC.

Prospective, randomized, double-blinded trial of bone marrow cell transplantation combined with coronary surgery - perioperative safety study.

We present here a sub-study of our prospective, randomized, double-blinded trial of bone marrow mononuclear cell (BMMC) transplantation with coronary artery bypass surgery (CABG) (ClinicalTrials.gov Identifier: NCT00418418), evaluating our secondary end-point concerning hospital stay as well as perioperative morbidity. Injecting a substantial amount of biologically active cells into a diseased myocardium inspires concerns for safety, a concern overlooked in previous trials. We evaluated the immediate perioperative effects of intramyocardial injection of autologous BMMCs combined with CABG. In a randomized double-blinded manner, 39 patients received injections either of BMMCs (n = 20) or of vehicle medium (n = 19). The patients' haemodynamics, arterial blood gases, systemic vein oxygen level, blood glucose, acid-base balance, lactate, haemoglobin, body temperature and diuresis, as well as medications needed, were recorded in the operating theatre and in the intensive care unit (ICU) every 4 h throughout the first postoperative 24 h. No dissimilarities in these parameters were detectable. In the ICU, the median need for adrenaline was 0.0086 µg/kg/min (first quartile 0.0000, third quartile 0.0204) for controls and 0.0090 µg/kg/min (0.0000, 0.0353) for BMMC patients (P = 0.757); for noradrenaline, 0.0586 µg/kg/min (0.0180, 0.0888) for controls and 0.0279 µg/kg/min (0.0145, 0.0780) for BMMC patients (P = 0.405). The median stay at the ICU was 2 days for both groups (1, 2 for controls; 1, 3 for BMMCs; P = 0.967). Within the first postoperative day, one control patient had an elevated level of creatine kinase-myocardial band fraction mass (CK-MBm) up to >100 µg/l; no BMMC patient showed elevated CK-MBm levels (P = 0.474). Both intramyocardial BMMC and placebo injections appear safe during surgery and immediate ICU stay after treatment of heart failure.

Intravenous administration of bone marrow mononuclear cells alleviates hearing loss after transient cochlear ischemia through paracrine effects

Bone marrow mononuclear cells (BMMCs) are known to enhance recovery from ischemic insults by secreting angiogenic factors and inducing the expression of angiogenic factors from host tissues. Therefore, the transplantation of BMMCs is considered a potential approach to promoting the repair of ischemic damaged organs. Here, we investigated the influence of BMMCs on progressive hair cell degeneration after transient cochlear ischemia in gerbils. Transient cochlear ischemia was produced by extracranial occlusion of the bilateral vertebral arteries immediately before their entry into the transverse foramen of the cervical vertebra. An intravenous injection of BMMCs prevented ischemia-induced hair cell degeneration and ameliorated hearing impairment. A tracking study showed that BMMCs injected into the femoral vein were limited in the spiral artery of the cochlea, suggesting that, although transplanted BMMCs were retained within the spiral ganglion area of the cochlea, they were neither transdifferentiated into cochlear cells nor fused with the injured hair cells and supporting cells in the organ of Corti to restore their functions. We also showed that the protein level of neurotrophin-3 and glial cell line-derived neurotrophic factor in the organ of Corti was upregulated after treatment with BMMCs. These results suggested that BMMCs have therapeutic potential possibly through paracrine effects. Thus, we propose the use of BMMCs as a potential new therapeutic strategy for hearing loss.

CCN1 enhances angiogenic potency of bone marrow transplantation in a rat model of hindlimb ischemia.

Implantation of autologous bone marrow mononuclear cells (BM-MNCs) has been performed in ischemic tissues, for stimulation of angiogenesis, but the limited number of BM-MNCs in patients with hindlimb ischemia disease may offset their overall therapeutic efficacy. CCN1 is a novel and essential regulator during angiogenesis. We evaluated whether CCN1 and BM-MNC are capable of promoting angiogenesis in hindlimb ischemia. In this study, we created the rat model of hindlimb ischemia, and then the rats were randomly divided into four groups: CCN1 infusion plus BM-MNC transplantation (CCN1 + BM-MNCs group), CCN1 infusion plus PBS injection (CCN1 group), vehicle infusion plus BM-MNC transplantation (BM-MNCs group) and vehicle infusion plus PBS injection (control group). The combination of CCN1 and BM-MNC therapy could increase blood perfusion, capillary/muscle fiber ratio and tissue oxygenation in ischemic hindlimb. Moreover, CCN1 could not only inhibit the apoptosis of BM-MNCs, but also enhance the adhesiveness of BM-MNCs to HUVEC. Taken together, CCN1 enhanced angiogenesis of BM-MNC transplantation, and combining CCN1 with BM-MNC transplantation is a useful alternative for ischemic limbs.