Bone Marrow-derived Mononuclear Cells Research Articles

A mouse model of osteonecrotic femoral head induced by methylprednisolone and liposaccharide

Introduction: Osteonecrosis of the femoral head is caused by various factors, including prolonged use of steroid drugs, use of alcohol, vascular injuries and hemoglobinopathies. This study aims to develop a mouse model for glucocorticoid-induced avascular necrosis (AVN) of the femoral head. Methods: Adult mice were randomly divided into two groups: experimental and control. Group A (the experimental group) was given (via intramuscular injection) 10 mg/kg of lipopolysaccharide (LPS) and 30 mg/kg of methylprednisolone (MPS). Each mouse additionally received MPS in divided oral doses of 13 mg/kg for 10 consecutive days. Group B (the control group) received normal saline at the same location and same volume as those in Group A. Histological changes of the femoral heads were observed by electron microscopy at 3, 5, and 7 weeks after the last chemical injection. The percentage of empty lacunae was measured randomly and the expression of fibrocartilage was evaluated using an image analyz­ing system. The expression of CD31 and VEGF-R2 were observed by immunohistochemistry. The bone marrow-derived mononuclear cells were stained with propidium iodide and cell cycle was analyzed by flow cytometry. Results: The results showed that at weeks 3 and 5, mice in Group A showed an increase in body weight. From weeks 5 to 7, mouse body weight in both groups remained constant. No difference in bone morphology was observed at week 7. The percentage of empty lacunae was 5.87 2.49% at week 5 and 21.58 8.10% at week 7. After 7 weeks, chondrocyte degeneration and fibrocartilage expression were observed. Moreover, the density of CD31 and VEGF-R2 markers increased in the femoral head. The rate of apoptosis in the bone marrow increased at week 3 then decreased. Conclusion: The data show that MPS, combined with LPS, can induce in mice features typical of early AVN of the femoral head.

Progesterone modulates endothelial progenitor cell (EPC) viability through the CXCL12/CXCR4/PI3K/Akt signalling pathway.

Progesterone treatment can effectively increase levels of circulating endothelial progenitor cells (EPCs) and improve neurological functional outcome in a traumatic brain injury (TBI) rat model. However, the mechanisms of progesterone's effects on EPC viability remain elusive. The CXCL12/CXCR4 (CXC chemokine ligand 12/CXC chemokine receptor 4) signalling pathway regulates cell proliferation; we hypothesize that it mediates progesterone-induced EPC viability. EPCs were isolated from bone marrow-derived mononuclear cells (BM-MNCs) and treated with progesterone (5, 10 and 100 nm). MTS assay was used to investigate EPC viability. Protein expression was examined by Western blotting, ELISA assay and flow cytometry. Cell membrane and cytoplasm proteins were extracted with membrane and cytoplasm protein extraction kits. CXCR4 antagonist (AMD3100) and phosphatidylinositol 3-kinases (PI3K) antagonist (LY294002) were used to characterize underlying mechanisms. Progesterone-induced EPC viability was time- and dose-dependent. Administration of progesterone facilitated EPC viability and increased expression of CXCL12 and phosphorylated Akt (also known as protein kinase B, pAkt) activity (P < 0.05). Progesterone did not regulate CXCR4 protein expression in cultured EPC membranes or cytoplasm. However, progesterone-induced EPC viability was significantly attenuated by AMD3100 or LY294002. Inhibition of the signalling pathway with AMD3100 and LY294002 subsequently reduced progesterone-induced CXCL12/CXCR4/PI3K/pAkt signalling activity. The CXCL12/CXCR4/PI3K/pAkt signalling pathway increased progesterone-induced EPC viability.

Chronic Myelocytic Leukemia (CML) Patient-Derived Dendritic Cells Transfected with Autologous Total RNA Induces CML-Specific Cytotoxicity.

The oncogenic bcr/abl1 fusion gene is a chronic myelogenous leukemia (CML)-specific antigen which is absent in normal tissues. This makes bcr/abl1 a perfect target for developing CML vaccines that elicit specific immune responses against minimal residual disease while sparing normal tissue. The aim of this study was to use different methods to induce dendritic cells (DCs) derived from patients with CML (CML-DCs) and analyze them for CML-specific tumor cytotoxicity for immune therapy. Bone marrow-derived mononuclear cells from ten CML patients were studied to induce CML-DC differentiation in the presence of recombinant human interleukin-4, rh-granulocyte-macrophage-colony stimulating factor, and tumor necrosis factor-alpha with either a total RNA-lipofectamine complex, total RNA or CML tumor lysate (freeze-thawed). CML-DC maturation, confirmed by expression of CD1α, CD40, CD80, CD83, CD86 and by real-time polymerase chain reaction, validated the CML-origin of these DC cells. CML-DCs stimulated cytotoxic T-cell (CTL) apoptosis, high levels of IL-12 secretion, and had significant inhibitory effect on K562 tumorigenicity in nude mice. CML-DCs pulsed with total RNA by lipofectamine transfection produced the strongest effect in tumor-specific CTL functions. These results indicate that CML-DCs transfected with total RNA by lipofectamine induce the strongest CTL cytotoxicity and have the greatest potential for CML immune therapy. This study holds promise for a DC-based strategy for inducing anti-leukemia responses and establishes a foundation for developing RNA vaccination against CML.

Comparison of Bone Marrow-derived Mononuclear Cells vs. Mesenchymal Stem Cells in Protecting Random-pattern Skin Flap Survival in Rats

Both cultured Bone Marrow Mesenchymal Stem Cells (BMSCs) and non-cultured Bone Marrow Mononuclear Cells (BM-MNCs) can improve random-pattern skin flap survival. However, whether prior culture expansion of bone marrow cells is beneficial for this therapy remains unclear. In the current study, the protective effects of BM-MNCs and BMSCs derived from identical bone marrow aspirates were compared in a random-pattern skin flap rat model. The mean skin flap survival rates were 71.6 ± 8.4% in the BM-MNC-treated group and 66.2 ± 3.1% in the BMSCtreated group, both of which were significantly higher than the control group (55.9 ± 3.4%). The protective effects were confirmed by blood perfusion analysis and vessel density assay. However, no significant difference was observed between the cell transplanted groups. These results indicate that the current method for pre-culture of BMSCs does not bring therapeutic benefits in skin flap protection. Therefore, BM-MNCs without pre-culture might be more practicable in the clinical setting.

Treatment of Large Bone Defects with a Vascularized Periosteal Flap in Combination with Biodegradable Scaffold Seeded with Bone Marrow-Derived Mononuclear Cells: An Experimental Study in Rats.

The surgical treatment of large bone defects continues to pose a major challenge in modern trauma and orthopedic surgery. In this study we test the effectiveness of a tissue engineering approach, using three-dimensional (3D) β-tricalcium phosphate (β-TCP) scaffolding plus bone marrow-derived mononuclear cells (BM-MNCs), combined with a vascularized periosteal flap, in a rat femur critical size defect model. Eighty rats were randomly allocated into four equal groups. Under general anesthesia, critical size defects were created on their femurs and were treated with (1) Vascularized periosteal flap alone, (2) Vascularized periosteal flap+β-TCP scaffolding, (3) Vascularized periosteal flap+β-TCP scaffolding+ligated vascular pedicle, and (4) Vascularized periosteal flap+β-TCP scaffolding+BM-MNCs. After 4 and 8 weeks animals were euthanized and the bone defects were harvested for analysis of new bone formation, vascularization, and strength using histology, immunohistology, micro-CT, and biomechanical testing, respectively. Group 1: (P. flap) Increase in new bone formation and vascularization. Group 2: (P. flap+scaffold) Increase in new bone formation and vascularization. Group 3: (P. flap+scaffold+ligated vascular pedicle) No new bone formation and no vascularization. Group 4: (P. flap+scaffold+BM-MNCs) A significant (p < 0.05) increase was seen in new bone formation, vascularization, and strength in bones treated with flaps, scaffold, and BM-MNCs, when compared with the other treatment groups. Combining a vascularized periosteal flap with tissue engineering approach (β-TCP scaffolding and BM-MNC) results in significantly improved bone healing in our rat femur critical size bone defect model.

Intraportal Infusion of Bone Marrow Mononuclear or CD133+ Cells in Patients With Decompensated Cirrhosis: A Double-Blind Randomized Controlled Trial.

The present study assessed the effects of intraportal infusions of autologous bone marrow-derived mononuclear cells (MNCs) and/or CD133+ cells on liver function in patients with decompensated cirrhosis. We randomly assigned 27 eligible patients to a placebo, MNCs, and/or CD133+ cells. Cell infusions were performed at baseline and month 3. We considered the absolute changes in the Model for End-Stage Liver Disease (MELD) scores at months 3 and 6 after infusion as the primary outcome. The participants and those who assessed the outcomes were unaware of the treatment intervention assignments. After 6 months, 9 patients were excluded because of liver transplantation (n=3), hepatocellular carcinoma (n=1), loss to follow-up (n=3), and death (n=2). The final analysis included 4 patients from the CD133+ group, 8 from the MNC group, and 6 from the placebo group. No improvement was seen in the MELD score at month 6 using either CD133+ cells or MNC infusions compared with placebo. However, at month 3 after infusion, a trend was seen toward a higher mean absolute change in the MELD score in patients who had received CD133+ cells compared with placebo (-2.00±1.87 vs. -0.13±1.46; p=.08). No significant adverse events occurred in the present study. A transient improvement in the MELD score was observed in subjects treated with CD133+ cells but not in the MNC or placebo group. Although the study was not powered to make definitive conclusions, the data justify further study of CD133+ therapy in cirrhotic patients. Cell therapy is a new approach in liver disease. Several clinical experiments have been reported on the safety of bone marrow-derived stem cells to treat liver disorders. However, the effectiveness of these approaches in the long-term follow-ups of patients initiated controversial discussions among the scientific community. A double-blind randomized controlled trial was designed to address this concern scientifically. A transient improvement in the patients' signs occurred; however, for a sustainable result, more work is needed. The results of multiple administrations of cells reported in the present study can be compared with the results from other single-injection studies.

Abstract 18016: Cell and Hydrogel-based Therapies Employing Cathelicidin Related Antimicrobial Peptide (CRAMP) Attenuate Cardiac Dysfunction in a Mouse Model of Myocardial Infarction: Potential Therapeutic Targets in Ischemic Heart Disease

Introduction: Limited stem cell retention following intracoronary administration has reduced the clinical efficacy of this novel therapy. Cathelicidins related antimicrobial proteins (CRAMPs) have been shown to prime bone marrow Derived mononuclear Cells (BMMNC) migration towards low gradients of SDF-1 suggesting a potential role in BMMNC retention. Here, we assessed the therapeutic efficacy of CRAMP in the context of BMMNC recruitment and retention via intracardiac delivery of CRAMP-treated BMMNCs or CRAMP-packed hydrogels (HG) post-myocardial infarction (MI). Methods and Results: During the in vivo cell studies, mice were randomized into 3 groups: MI followed by injection of PBS, BMMNCs alone, and BMMNCs incubated with CRAMP. During the in vivo HG studies, BM GFP chimera mice were randomized into 4 groups: MI followed by injection of HG alone, HG + SDF-1, HG + SDF-1 + CRAMP, HG + CRAMP. Changes in infarct size and cardiac function between 5 days and 5 weeks after MI were assessed using cardiac MRI (CMR) and echocardiography respectively. Treatment of BMMNCs with CRAMP enhanced their migration towards low, yet physiological, levels of SDF-1 in vitro (Fig 1A). In vivo, BMMNC+CRAMP therapy was associated with significant increase in capillary density as compared to the BMMNC-alone and control groups (Fig 1B). Moreover, BMMNC+CRAMP administration led to improvement in cardiac function and reduction in infarct size as compared to other groups (Fig 1C-D). Similar to the BMMNCs+CRAMP cell therapy, HG + SDF-1 + CRAMP administration post-MI led to improvement of cardiac function and reduction in infarct size compared with other groups (Fig 1E-F). Conclusion: Cathelicidins enhance BMMNC retention and recruitment after intramyocardial administration post-MI resulting in improvements in heart physiology and recovery. Therapies employing these strategies may represent an attractive method for improving outcomes of regenerative therapies in human studies.

Abstract 17510: The First Prediction Model for Response to Autologous Bone Marrow-derived Mononuclear Cells After Myocardial Infarction Based on the REPAIR-AMI Trial

Background: Administration of autologous bone marrow-derived mononuclear cells (BM-MNC) resulted in favorable outcomes after myocardial infarction (MI) compared to placebo in the REPAIR-AMI trial. Pre-clinical studies have shown that individual risk factors negatively influence cell function. To date, it is not known how these risk factors modify treatment effect in clinical trials and if potential (non-)responders can be identified. Aim: To investigate the effect of individual risk factors on functional outcome after BM-MNC therapy and to establish a prediction model for treatment response in patients with MI. Methods: Data from the REPAIR-AMI trial were used, consisting of 186 patients who had complete baseline and follow-up measurements. We performed univariable and multivariable linear regression with 18 predefined baseline characteristics using the difference in baseline EF and after 4 months (ΔEF4) as primary outcome. Our main goal was to identify interaction terms of predictors with BM-MNC treatment; i.e. effect modifiers of treatment response. An individual estimate of treatment effect over placebo (ΔΔEF4) was created by extrapolating an ‘untreated’ reference value for ΔEF4, based on the placebo-arm and comparing it to the observed value after treatment. Treatment response was defined as a ΔΔEF4 of &gt;5% (so 5% over a predicted placebo value). Discrimination was quantified by the area under the ROC-curve (AUC). Results: The predictors age, weight, baseline EF and ESV showed an interaction with cell treatment in multivariable analysis with backward selection. Subsequently, the prediction model for individual ΔΔEF4 included age (-0.18/year, p=0.008), weight (+0.16/kg, p=0.01), baseline EF (-0.46/%, p=0.0001) and ESV (-0.09/ml, p=0.06). The predictive capacity for response to therapy showed an AUC of 79.2% (95% CI 69.7-88.6), which retained 74.8% (95% CI 65.0-84.5) after shrinkage. Conclusion: Response to BM-MNC administration after MI is influenced by age, weight, and baseline cardiac parameters. Using these continuous predictors, potential treatment responders can be accurately identified. Interestingly, an adverse risk-factor profile was associated with greater response, with potential implications for future patient selection.

Abstract 17998: Impairment of Vascular Bone Marrow Niche Predicts Death in Patients With Chronic Heart Failure

Background: The bone marrow vascular niche is essential for hematopoietic regeneration. Patients with chronic heart failure are characterized by myelosuppresion and anemia. Therefore, in order to investigate whether chronic heart failure is associated with an impairment in the vascular niche of the bone marrow, we measured in bone marrow plasma levels of the prototypical angiogenic factor PlGF (placental growth factor), as well as, its soluble inhibitor, sFlt-1 (Fms-like tyrosine kinase 1), which acts as a decoy receptor for PlGF, as a measure of the functional integrity of the vascular bone marrow niche. Methods and Results: Bone marrow plasma levels of sFlt-1 and PlGF were determined by ELISA in 187 patients (mean age: 62.2±11.3years) with chronic heart failure, who were enrolled in the Frankfurt Bone Marrow-Derived Mononuclear Cell Therapy Registry. Fifty-one patients died within the 5-year follow-up period. Patients with increased bone marrow-derived sFlt-1/PlGF ratio had a 1.35-fold increased risk for death (95%CI: 1.08 to 1.68, p=0.007). This association was robust after adjustment for age, gender, left ventricular ejection fraction (LVEF), eGFR and NT-proBNP serum levels (HR:1.3, 95%CI: 1.014 to 1.678, p=0.038) or the Seattle Heart Failure score (HR:1.38, 95%CI: 1.112 to 1.723, p=0.004). Importantly, this association remained significant even after adjustment for colony-forming unit capacity (HR:1.33, 95%CI: 1.051 to 1.675, p=0.018) and SDF-1-induced migratory capacity of bone marrow-derived cells (HR:1.35, 95%CI: 1.087 to 1.678, p=0.007). Conclusions: The present study for the first time discloses that an impairment of the angiogenic potential of the bone marrow microenvironment, defined as increased sFlt-1 to PlGF ratio in the bone marrow plasma, is independently associated with mortality in patients with chronic heart failure. These data further corroborate the importance of the bone marrow-heart axis to predict clinical outcome in patients with chronic heart failure.

Autologous Bone Marrow Mononuclear Cells Exert Broad Effects on Short- and Long-Term Biological and Functional Outcomes in Rodents with Intracerebral Hemorrhage.

Autologous bone marrow-derived mononuclear cells (MNCs) are a potential therapy for ischemic stroke. However, the effect of MNCs in intracerebral hemorrhage (ICH) has not been fully studied. In this study, we investigated the effects of autologous MNCs in experimental ICH. ICH was induced by infusion of autologous blood into the left striatum in young and aged male Long Evans rats. Twenty-four hours after ICH, rats were randomized to receive an intravenous administration of autologous MNCs (1 × 10(7) cells/kg) or saline. We examined brain water content, various markers related to the integrity of the neurovascular unit and inflammation, neurological deficit, neuroregeneration, and brain atrophy. We found that MNC-treated young rats showed a reduction in the neurotrophil infiltration, the number of inducible nitric oxide synthase-positive cells, and the expression of inflammatory-related signalings such as the high-mobility group protein box-1, S100 calcium binding protein B, matrix metalloproteinase-9, and aquaporin 4. Ultimately, MNCs reduced brain edema in the perihematomal area compared with saline-treated animals at 3 days after ICH. Moreover, MNCs increased vessel density and migration of doublecortin-positive cells, improved motor functional recovery, spatial learning, and memory impairment, and reduced brain atrophy compared with saline-treated animals at 28 days after ICH. We also found that MNCs reduced brain edema and brain atrophy and improved spatial learning and memory in aged rats after ICH. We conclude that autologous MNCs can be safely harvested and intravenously reinfused in rodent ICH and may improve long-term structural and functional recovery after ICH. The results of this study may be applicable when considering future clinical trials testing MNCs for ICH.

Anticoagulants on yield of bone marrow-derived mononuclear cells harvested from dogs

Cell therapy with bone marrow-derived mononuclear cells is an alternative to therapy with mesenchymal stem cell cultures. The aim of the present research was the comparison of the yield of bone marrow-derived mononuclear cells harvested from dogs with two different anticoagulants. Bone marrow was harvested from the iliac crest of five healthy dogs aged between 15 and 30 months, and the effect of two anticoagulant solutions, CPDA-1 (citrate phosphate dextrose adenine-1) and heparin, on the isolation of mononuclear cells was compared. Mononuclear cells were isolated in a density gradient and stained for CD9 and CD44 for characterization by flow cytometry. Means were compared using Student's paired t-test. Samples harvested with CPDA-1 yielded an average of 5.16x106 (±1.76x106) to 20.20x106 (±1.55x106) mononuclear cells/mL, whereas the yield of samples harvested with heparin varied between 4.56x106 (±0.69x106) and 24.30x106 (±2.12x106) mononuclear cells mL-1. By flow cytometry, mean percentage of double-stained cells varied from 1.96% (±0.64%) to 5.01% (±0.73%) for CPDA-1 and from 2.23% (±0.70%) to 7.27% (±0.97%) for heparin. No significant statistical differences were observed on yield or CD9 and CD44 expression. Further studies are recommended to assess efficacy of CPDA on mononuclear cell isolation.

Improved outcome with repeated intracoronary injection of bone marrow-derived cells within a registry: rationale for the randomized outcome trial REPEAT.

Regenerative therapies have evolved as a promising new option in the treatment of post-infarction heart failure. A major limitation of intracoronary application of autologous bone marrow-derived mononuclear cells (BM-MNCs) is that homing of the applied cells is profoundly reduced in patients with post-infarction heart failure compared with patients with acute myocardial infarction. However, early pilot and also randomized controlled trials have demonstrated significant improvements in overall cardiac function. The aim of the present analysis was to quantify a potential mortality risk reduction and reduced hospitalization in order to provide data for a prospective outcome trial. The results of an ongoing single-centre registry including 297 post-infarction heart failure patients suggest that repeated intracoronary application of autologous bone marrow-derived cells is associated with a significant better 2-year survival compared with a single BM-MNC application (2-year survival 93.6 vs. 84.0%, P = 0.03). Likewise, mortality is significantly lower at 2-year follow-up compared with the mortality estimated by the use of the Seattle Heart Failure Model (SHFM) in patients receiving repeated BM-MNC application (observed mortality 6.4%, predicted mortality 16.2%, P = 0.02). Although the trend persisted at 3-year follow-up, the mortality reduction was no longer statistically significant between single and repeated treatment (mortality 21.9 vs. 13.7%, P = 0.06). Repeated intracoronary administration of BM-MNC appears to be associated with improved clinical outcome compared with single treatment at 2 years. This registry provides the rationale for the design of the multicentre randomized, controlled, open-label REPEAT trial, which prospectively compares the effects of single vs. repeated intracoronary application of autologous BM-MNC on total and SHFM-predicted mortality in patients with chronic post-infarction heart failure.

Quality of Life After Treatment with Autologous Bone Marrow Derived Cells in No Option Severe Limb Ischemia

Quality of life (QoL) is an important outcome in evaluating treatment effect in severe limb ischemia. The randomized, double blind, placebo controlled JUVENTAS trial, investigating the effect of bone marrow derived mononuclear cell (BMMNC) administration in no option severe limb ischemia, showed an improved QoL at 6 months compared with baseline in both the treatment and placebo groups. The aim of the present study was to evaluate whether the improved QoL persisted beyond 6 months' follow up, whether this differed in both trial arms, and if major amputation influenced QoL. Short form 36 (SF-36) and EuroQol 5D (EQ5D), including the EQ Visual Analogue Scale (EQ-VAS), questionnaires were sent to JUVENTAS trial participants. In the JUVENTAS trial, a norm based scoring method was applied to report the results of the SF-36. The results of the long-term follow up were compared with baseline and 6 month follow up and the results of both trial arms were compared, as were the results of patients with and without amputation. One hundred and nine patients (86.5% of surviving patients) responded to the questionnaires. Median follow up after inclusion was 33 months (interquartile range [IQR] 21.2-50.6) for the BMMNC and 36 months (IQR 21.4-50.9) for the placebo group. The improvement in QoL at 6 months persisted in both arms at a median follow up of 35 months. The long-term QoL did not differ between the BMMNC and placebo group in any of the SF-36 or EQ5D domains. Patients with and without a major amputation had similar QoL scores. The increased QoL in patients with no option severe limb ischemia persisted until 3 years after inclusion, but did not differ between the BMMNC and placebo arms or between patients with and without a major amputation.

Effect of cell seeding on neotissue formation in a tissue engineered trachea

BackgroundSurgical management of long segment tracheal disease is limited by a paucity of donor tissue and poor performance of synthetic materials. A potential solution is the development of a tissue-engineered tracheal graft (TETG) which promises an autologous airway conduit with growth capacity. MethodsWe created a TETG by vacuum seeding bone marrow-derived mononuclear cells (BM-MNCs) on a polymeric nanofiber scaffold. First, we evaluated the role of scaffold porosity on cell seeding efficiency in vitro. We then determined the effect of cell seeding on graft performance in vivo using an ovine model. ResultsSeeding efficiency of normal porosity (NP) grafts was significantly increased when compared to high porosity (HP) grafts (NP: 360.3±69.19×103 cells/mm2; HP: 133.7±22.73×103 cells/mm2; p<0.004). Lambs received unseeded (n=2) or seeded (n=3) NP scaffolds as tracheal interposition grafts for 6weeks. Three animals were terminated early owing to respiratory complications (n=2 unseeded, n=1 seeded). Seeded TETG explants demonstrated wound healing, epithelial migration, and delayed stenosis when compared to their unseeded counterparts. ConclusionVacuum seeding BM-MNCs on nanofiber scaffolds for immediate implantation as tracheal interposition grafts is a viable approach to generate TETGs, but further preclinical research is warranted before advocating this technology for clinical application.

Amelioration of Autism by Autologous Bone Marrow Mononuclear Cells and Neurorehabilitation: A Case Report

Autism is a clinically and etiologically heterogeneous disorder characterized by deficits in social interaction, communication, behavior and cognitive skills. The etiological basis of autism still remains poorly understood despite several attempts to decipher its neuropathology from different perspectives. Presently available treatment modalities address only limited autism-associated symptoms and are at best palliative. In this report, we present the case of a 7 year old boy with autism treated with intrathecal administration of autologous bone marrow derived mononuclear cells (BMMNCs). On regular follow ups conducted at 3 and 6 months post-treatment, clinically significant behavioral, social, communication and cognitive improvements were reported. These findings were well supported by objective improvements on the Indian Scale of Assessment of Autism (ISAA), Childhood Autism Rating Scale (CARS), Clinical Global Impression (CGI) and Pediatric Functional Independence Measure (WeeFIM). The ISAA score improved from 131 to 112, CARS improved significantly from 40.5 (severely autistic) to 32 (mild to moderate autism), along with an improved WeeFIM score from 31 to 36. Severity of illness on CGI (CGI I) changed from 4 (moderately ill) to 3 (mildly ill). Global improvement on CGI (CGI II) was measured at a score of 2 (much improved), along with an efficacy index (CGI III) of 5 showing moderate therapeutic effect. No adverse events were reported throughout the course of the treatment. Through this case report, we demonstrate that treatment with autologous BMMNCs is safe, feasible and has the potential to ameliorate autism.

Editorial: Stem cells and progenitor cells in ischemic stroke—fashion or future?

Editorial: Stem cells and progenitor cells in ischemic stroke—fashion or future?

Bisphenol A (BPA) stimulates the interferon signaling and activates the inflammasome activity in myeloid cells

Environmental factors contribute to the development of autoimmune diseases, including systemic lupus erythematosus (SLE), which exhibits a strong female bias (female-to-male ratio 9:1). However, the molecular mechanisms remain largely unknown. Because a feedforward loop between the female sex hormone estrogen (E2) and type I interferon (IFN-α/β)-signaling induces the expression of certain p200-family proteins (such as murine p202 and human IFI16) that regulate innate immune responses and modify lupus susceptibility, we investigated whether treatment of myeloid cells with bisphenol A (BPA), an environmental estrogen, could regulate the p200-family proteins and activate innate immune responses. We found that treatment of murine bone marrow-derived cells (BMCs) and human peripheral blood mononuclear cells with BPA induced the expression of ERα and IFN-β, activated the IFN-signaling, and stimulated the expression of the p202 and IFI16 proteins. Further, the treatment increased levels of the NLRP3 inflammasome and stimulated its activity. Accordingly, BPA-treatment of BMCs from non lupus-prone C57BL/6 and the lupus-prone (NZB×NZW)F1 mice activated the type I IFN-signaling, induced the expression of p202, and activated an inflammasome activity. Our study demonstrates that BPA-induced signaling in the murine and human myeloid cells stimulates the type I IFN-signaling that results in an induction of the p202 and IFI16 innate immune sensors for the cytosolic DNA and activates an inflammasome activity. These observations provide novel molecular insights into the role of environmental BPA exposures in potentiating the development of certain autoimmune diseases such as SLE.

1α,25-Dihydroxyvitamin D3 inhibits the differentiation and bone resorption by osteoclasts generated from Wistar rat bone marrow-derived macrophages.

The steroid hormone 1α,25-dihydroxyvitamin D3 [1α,25-(OH)2D3] plays an important role in maintaining a balance in calcium and bone metabolism. To study the effects of 1α,25-(OH)2D3 on osteoclast (OC) formation and bone resorption, OC differentiation was induced in bone marrow-derived mononuclear cells from Wistar rats with the addition of macrophage colony stimulating factor and receptor activator for nuclear factor-κB ligand in vitro. Cells were then treated with 1α,25-(OH)2D3 at 10-9, 10-8 or 10-7 mol/l. OCs were identified using tartrate-resistant acid phosphatase staining and activity was monitored in the absorption lacunae by scanning electron microscopy. Expression levels of functional proteins associated with bone absorption, namely carbonic anhydrase II, cathepsin K and matrix metalloproteinase-9 were evaluated by western blot analysis. The results showed that 1α,25-(OH)2D3 inhibited the formation and activation of OCs in a dose-dependent manner and downregulated the expression levels of bone absorption-associated proteins.

Injured hepatocyte-released microvesicles induce bone marrow-derived mononuclear cells differentiation

The ability of bone marrow-derived mononuclear cells (BMMCs) to differentiate into hepatocyte-like cells under different conditions has been demonstrated previously. In the present study, we investigated the effect of CCl4-injured hepatocytes on the differentiation of the non-adherent (NAD) fraction of BMMCs. Differentiation (cell fate) was analyzed after 2, 6 and 24h of co-culture by gene and protein expression and by urea production. We also evaluated the presence of microvesicles (MVs) in the supernatant of differentiated cells, their content and the ability of these cells to absorb them. Hepatocyte-like characteristics were observed in the NAD cells after 24h of co-culture with injured hepatocytes. Cells that were co-cultured with healthy hepatocytes did not present signs of differentiation at any analyzed time point. Analysis of the supernatant from differentiated cells revealed the presence of MVs carrying hepatocyte-specific mRNAs, including Albumin, Coagulation factor V, Alpha-fetoprotein, and Cytokeratin 18. The incorporation of injured hepatocyte-derived MVs by NAD cells was shown at 24h, suggesting a possible role for MVs in the induction of cell plasticity.

Pilot safety study of intrabronchial instillation of bone marrow-derived mononuclear cells in patients with silicosis

BackgroundSilicosis is an occupational disease for which no effective treatment is currently known. Systemic administration of bone marrow-derived mononuclear cells (BMDMCs) has shown to be safe in lung diseases. However, so far, no studies have analyzed whether bronchoscopic instillation of autologous BMDMCs is a safe route of administration in patients with silicosis.MethodsWe conducted a prospective, non-randomized, single-center longitudinal study in five patients. Inclusion criteria were age 18–50 years, chronic and accelerated silicosis, forced expiratory volume in 1 s <60 % and >40 %, forced vital capacity ≥60 % and arterial oxygen saturation >90 %. The exclusion criteria were smoking, active tuberculosis, neoplasms, autoimmune disorders, heart, liver or renal diseases, or inability to undergo bronchoscopy. BMDMCs were administered through bronchoscopy (2 × 107 cells) into both lungs. Physical examination, laboratory evaluations, quality of life questionnaires, computed tomography of the chest, lung function tests, and perfusion scans were performed before the start of treatment and up to 360 days after BMDMC therapy. Additionally, whole-body and planar scans were evaluated 2 and 24 h after instillation.ResultsNo adverse events were observed during and after BMDMC administration. Lung function, quality of life and radiologic features remained stable throughout follow-up. Furthermore, an early increase of perfusion in the base of both lungs was observed and sustained after BMDMC administration.ConclusionAdministration of BMDMCs through bronchoscopy appears to be feasible and safe in accelerated and chronic silicosis. This pilot study provides a basis for prospective randomized trials to assess the efficacy of this treatment approach.Clinical trials.gov identifierNCT01239862 Date of Registration: November 10, 2010