Autologous Bone Marrow Mesenchymal Stem Cells Research Articles

Improvement in Left Ventricular Function with Intracoronary Mesenchymal Stem Cell Therapy in a Patient with Anterior Wall ST-Segment Elevation Myocardial Infarction

Background/AimsThe progression and development of congestive heart failure is still considered a large problem despite the existence of revascularization therapies and optimal, state-of-the-art medical services. An acute myocardial infarction (AMI) is a major cause of congestive heart failure, so researchers are investigating techniques to complement primary percutaneous coronary intervention (PCI) or thrombolytic therapy to prevent congestive heart failure after AMI.MethodsTwenty-six patients with successful PCI for acute ST-segment elevation anterior wall myocardial infarction were assigned to either a control group (n = 12) or a bone marrow mesenchymal stem cells (BM-MSC) group (n = 14). The control group received optimum post-infarction treatment, and the BMSC group received intracoronary delivery of autologous BMSC at 1 month after PCI with the optimum medical treatment. The primary endpoint was a left ventricular ejection fraction (LVEF) change from baseline to 4-month follow-up, as determined via myocardial single-photon emission computed tomography (SPECT).ResultsThe global LVEF at baseline (determined 3.5 ± 1.5 days after PCI) was 35.4 ± 3.0% in the control group and 33.6 ± 4.7% in the BM-MSC group. BMSC transfer enhanced left ventricular systolic function primarily in anterior wall myocardial segments adjacent to the LAD infarcted area. Four months later, via SPECT, global LVEF had increased by 4.8 ± 1.9% in the control group and 8.8 ± 2.9% in the BM-MSC group (p = 0.031). The cell transfer did not increase the risk of adverse clinical events, in-stent restenosis, or proarrhythmic effects. The echocardiographic evaluation also revealed a significant increase in the LVEF value from baseline to the 4-month (9.0 ± 4.7 and 5.3 ± 2.6%, p = 0.023) and 12-month (9.9 ± 5.2% and 6.5 ± 2.7%, p = 0.048) follow-up in the BM-MSC group but not in the control group.ConclusionsIntracoronary administration of autologous BM-MSC was tolerable and safe with significant improvement in LVEF at 4-month (SPECT and echocardiography result) and 12-month (echocardiography result only) follow-up in patients with anterior AMI.

Bone marrow mesenchymal stem cells do not enhance intra-synovial tendon healing despite engraftment and homing to niches within the synovium

BackgroundIntra-synovial tendon injuries display poor healing, which often results in reduced functionality and pain. A lack of effective therapeutic options has led to experimental approaches to augment natural tendon repair with autologous mesenchymal stem cells (MSCs) although the effects of the intra-synovial environment on the distribution, engraftment and functionality of implanted MSCs is not known. This study utilised a novel sheep model which, although in an anatomically different location, more accurately mimics the mechanical and synovial environment of the human rotator cuff, to determine the effects of intra-synovial implantation of MSCs.MethodsA lesion was made in the lateral border of the lateral branch of the ovine deep digital flexor tendon within the digital sheath and 2 weeks later 5 million autologous bone marrow MSCs were injected under ultrasound guidance into the digital sheath. Tendons were recovered post mortem at 1 day, and 1–2, 4, 12 and 24 weeks after MSC injection. For the 1-day and 1–2-week groups, MSCs labelled with fluorescent-conjugated magnetic iron-oxide nanoparticles (MIONs) were tracked with MRI, histology and flow cytometry. The 4, 12 and 24-week groups were implanted with non-labelled cells and compared with saline-injected controls for healing.ResultsThe MSCs displayed no reduced viability in vitro to an uptake of 20.0 ± 4.6 pg MIONs per cell, which was detectable by MRI at minimal density of ~ 3 × 104 cells. Treated limbs indicated cellular distribution throughout the tendon synovial sheath but restricted to the synovial tissues, with no MSCs detected in the tendon or surgical lesion.The lesion was associated with negligible morbidity with minimal inflammation post surgery. Evaluation of both treated and control lesions showed no evidence of healing of the lesion at 4, 12 and 24 weeks on gross and histological examination.ConclusionsUnlike other laboratory animal models of tendon injury, this novel model mimics the failed tendon healing seen clinically intra-synovially. Importantly, however, implanted stem cells exhibited homing to synovium niches where they survived for at least 14 days. This phenomenon could be utilised in the development of novel physical or biological approaches to enhance localisation of cells in augmenting intra-synovial tendon repair.

Transcriptional Profiling of Mesenchymal Stem Cells Identifies Distinct Neuroimmune Pathways Altered by CNS Disease.

Background and ObjectivesBone marrow mesenchymal stem cells (BM-MSCs) are an attractive cell based therapy in the treatment of CNS demyelinating diseases such as multiple sclerosis (MS). Preclinical studies demonstrate that BM-MSCs can effectively reduce clinical burden and enhance recovery in experimental autoimmune encephalomyelitis (EAE), a commonly used animal model of MS. However, a number of recent clinical trials have not shown significant functional benefit following BM-MSC infusion into MS patients. One possibility for the discrepancy between animal and human studies is the source of the cells, as recent studies suggest BM-MSCs from MS patients or animals with EAE lack reparative efficacy compared to naïve cells. We sought to define important transcriptional and functional differences between diseased and naïve MSCs.Methods and ResultsWe utilized RNA Sequencing (RNA-Seq) to assess changes in gene expression between BM-MSCs derived from EAE animals and those derived from healthy controls. We show that EAE alters the expression of a large number of genes in BM-MSCs and changes in gene expression are more pronounced in chronic versus acute disease. Bioinformatic analysis revealed extensive perturbations in BM-MSCs in pathways related to inflammation and the regulation of neural cell development. These changes suggest that signals from EAE derived BM-MSCs inhibit rather than enhance remyelination, and in-vitro studies showed that conditioned medium from EAE MSCs fails to support the development of mature oligodendrocytes, the myelinating cells of the CNS.ConclusionsThese data provide insight into the failure of autologous BM-MSCs to promote recovery in MS and support the concept of utilizing non-autologous MSCs in future clinical trials.

Efficacy and long-term longitudinal follow-up of bone marrow mesenchymal cell transplantation therapy in a diabetic patient with recurrent lower limb bullosis diabeticorum

Bullosis diabeticorum is a rare presentation of cutaneous manifestation most commonly affecting the lower limbs in patients with diabetes. The appearance, often as insidious as its resolution, is characterized by tense blisters on the skin surfaces of the lower limbs and the feet. The cause still remains unclear, but it may relate to microangiopathy and neuropathy. In this report, we present a case of a 64-year-old male with multiple episodes of blistering in the left lateral lower limb after a traumatic fall who was subsequently diagnosed with type 2 diabetes mellitus. The patient had a history of poorly controlled blood glucose and subsequently developed vasculopathy and peripheral neuropathy. Despite appropriate glycemic control and antibiotics therapy, the patient developed recurrent bullosis diabeticorum on five separate occasions during a 2-year span from 2005 to 2007. Building on our success with ischemic diabetic foot, we used bone marrow mesenchymal stem cell (BMMSC) transplantation therapy for bullosis diabeticorum. After a 9-month treatment, this patient developed another episode of cellulitis in the same lower limb which was successfully treated with antibacterial therapy. It is interesting that the patient reported no recurrence in the next 10-year follow-up span. This study demonstrates that bullosis diabeticorum could appear even before the onset of diabetes, and vascular insufficiency predisposes to the occurrence of bullosis diabeticorum. Our findings suggest that autologous BMMSC transplantation therapy may be an effective measure for recurrent bullosis diabeticorum; however, this will require further investigation to be conclusive. Early identification of diabetes and its complications and appropriate treatment may improve clinical outcomes and prevent lower limb amputation.Trial registration: ClinicalTrials.gov Identifier: NCT00955669. Registered on August 10, 2009.

The safety and efficacy of magnetic targeting using autologous mesenchymal stem cells for cartilage repair.

A new cell delivery system using magnetic force, termed magnetic targeting, was developed for the accumulation of locally injected cells in a lesion. The aim of this study was to assess the safety and efficacy of mesenchymal stem cell (MSC) magnetic targeting in patients with a focal articular cartilage defect in the knee. MSC magnetic targeting for five patients was approved by the Ministry of Health Labour and Welfare of Japan. Autologous bone marrow MSCs were cultured and subsequently magnetized with ferucarbotran. The 1.0-T compact magnet was attached to a suitable position around the knee joint to allow the magnetic force to be as perpendicular to the surface of the lesion as possible. Then 1 × 107 MSCs were injected into the knee joint. The magnet was maintained in the same position for 10min after the MSC injection. The primary endpoint was the occurrence of any adverse events. The secondary endpoints were efficacy assessed by magnetic resonance imaging (MRI) T2 mapping and clinical outcomes using the International Knee Documentation Committee (IKDC) Subjective Knee Evaluation and the Knee Injury and Osteoarthritis Outcome Score (KOOS). No serious adverse events were observed during the treatment or in the follow-up period. Swelling of the treated knee joint was observed from the day after surgery in three of the five patients. The swelling resolved within 2weeks in two patients. MRI showed that the cartilage defect areas were almost completely filled with cartilage-like tissue. MOCART scores were significantly higher 48weeks postoperatively than preoperatively (74.8 ± 10.8 vs 27.0 ± 16.8, p = 0.042). Arthroscopy in three patients showed complete coverage of their cartilage defects. Clinical outcome scores were significantly better 48weeks postoperatively than preoperatively for the IKDC Subjective Knee Evaluation (74.8 ± 17.7 vs 46.9 ± 17.7, p = 0.014) and knee-related quality-of-life (QOL) in the KOOS (53.8 ± 26.4 vs 22.5 ± 30.8, p = 0.012). Magnetic targeting of MSCs was safely performed and showed complete coverage of the defects with cartilage-like tissues and significant improvement in clinical outcomes 48weeks after treatment. The magnetic targeting of MSCs is useful as a minimally invasive treatment for cartilage repair. IV.

Efficacy and Safety of Autologous Bone Marrow Mesenchymal Stem Cell Transplantation in Patients with Diabetic Retinopathy

Background/Aims: The treatment options for diabetic retinopathy (DR) are limited. Mesenchymal stem cells (MSCs) are a promising treatment option for diabetes and its complications. In this pilot clinical trial, we evaluated the safety and efficacy of intravenous autologous bone marrow MSCs (ABMSC) for the treatment of DR. Methods: In total, 34 eyes with non-proliferative or proliferative DR (NPDR, n = 19; PDR, n = 15) from 17 patients were analyzed. Treatment involved one intravenous infusion of 3 × 10⁶/kg ABSMCs. The patients’ vital signs were monitored, along with immune and allergic reactions. Treatment efficacy was evaluated via measurements of the following parameters at baseline, and at 1, 3, and 6 months after treatment: the levels of fasting blood glucose (FBG), Hemoglobin A1C (HbA_1C), interleukin-6 (IL-6), and hypersensitive C-reactive protein (CRP); best corrected visual acuity (BCVA); and central macular and subfield thickness (via optical computed tomography). Results: ABMSC infusion led to a significant decrease in FBG and CRP levels (P < 0.05). There were no significant differences in HbA_1C or IL-6 levels. Sub-group analysis revealed that only eyes in the NPDR group had the macular thickness reductions and a significant improvement in BCVA from baseline (P = 0.006 at 3 months and 0.027 at 6 months), while those in the PDR group did not. There were no acute reactions during the treatment or severe adverse events during the follow-up period. Conclusion: ABSMCs are a potentially safe and effective treatment option for DR, and the optimum therapeutic window appears to be during the NPDR stage.

Clinical Outcomes of Repeated Intraventricular Transplantation of Autologous Bone Marrow Mesenchymal Stem Cells in Chronic Haemorrhagic Stroke. A One-Year Follow Up

Object:Stroke, one of the most devastating diseases, is a leading cause of death and disability throughout the world and is also associated with emotional and economic problems. The main goal of this study was to investigate the clinical outcome of the intraventricular transplantation of bone marrow mesenchymal stem cells (BM-MSCs) in post-haemorrhagic stroke patients.Method:This study was done consisting of eight patients with supratentorial haemorrhagic stroke, who had undergone 24 weeks of standard treatment of stroke with stable neurological deficits. All of the patients received stem cell transplantation intraventricularly using autologous BM-MSCs. Six months and Twelve months after stem cells treatment, the clinical outcomes were measured using the National Institute of Health Stroke Scale (NIHSS) and adverse effect also observed.Result:The results of this study showed improvement of NIHSS score values before and after the treatment in five patients. No adverse effects or complications were detected during the 1-year observation.Conclusion: Intraventricular transplantation of BM-MSCs has shown benefits in improving the functional status of post-haemorrhagic stroke patients with no adverse effect.

Tissue-engineered composite scaffold of poly(lactide-co-glycolide) and hydroxyapatite nanoparticles seeded with autologous mesenchymal stem cells for bone regeneration.

A new therapeutic strategy using nanocomposite scaffolds of grafted hydroxyapatite (g-HA)/ poly(lactide-co-glycolide) (PLGA) carried with autologous mesenchymal stem cells (MSCs) and bone morphogenetic protein-2 (BMP-2) was assessed for the therapy of critical bone defects. At the same time, tissue response and in vivo mineralization of tissue-engineered implants were investigated. A composite scaffold of PLGA and g-HA was fabricated by the solvent casting and particulate-leaching method. The tissue-engineered implants were prepared by seeding the scaffolds with autologous bone marrow MSCs in vitro. Then, mineralization and osteogenesis were observed by intramuscular implantation, as well as the repair of the critical radius defects in rabbits. After eight weeks post-surgery, scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDX) revealed that g-HA/PLGA had a better interface of tissue response and higher mineralization than PLGA. Apatite particles were formed and varied both in macropores and micropores of g-HA/PLGA. Computer radiographs and histological analysis revealed that there were more and more quickly formed new bone formations and better fusion in the bone defect areas of g-HA/PLGA at 2-8 weeks post-surgery. Typical bone synostosis between the implant and bone tissue was found in g-HA/PLGA, while only fibrous tissues formed in PLGA. The incorporation of g-HA mainly improved mineralization and bone formation compared with PLGA. The application of MSCs can enhance bone formation and mineralization in PLGA scaffolds compared with cell-free scaffolds. Furthermore, it can accelerate the absorption of scaffolds compared with composite scaffolds.

Bone marrow mesenchymal stem cells combined with biomimetic hydrogel for repair of articular cartilage defect in porcine

Objective To investigate the feasibility of bone marrow mesenchymal stem cells (BMSCs) combined with type Ⅱ collagen- hyaluronic acid- oxidized chondroitin sulfate (Col Ⅱ-HA-OCS) biomimetic hydrogel to repair articular cartilage defect in porcine and the role of the transplanted cells played in the process of cartilage repair. Methods A articular cartilage defect model which remaining cartilage calcified zone was created in the knee of Bama minipigs, the autologous BMSCs was used as seeds for transplantation and was labeled by the 5, 6-carboxyfluorescein diacetate succinimidyl ester (CFDA SE). Animals were randomly divided into three groups: Group A (blank group) was left untreated, group B (cell-free biomimetic hydrogel group) was filled with biomimetic hydrogel and group C (BMSCs combined with biomimetic hydrogel group) was filled with the CFDA SE labeled autologous BMSCs combined with biomimetic hydrogel. One month after the operation, BrdU labeled liquid was injected intravenously into the animals 24 h and 48 h before specimens were taken from the executed animals. Partial cartilage repaired tissue in Group C was taken, cryosectioned and stained with DAPI and BrdU immunofluorescence. Confocal laser scanning microscope was used to observe and count the cells. Specimens of the three groups were analyzed through gross observation and histological staining, and scored according to the international cartilage repair society (ICRS) gross morphological score and ICRS histological score. Results Laser scanning confocal microscopy showed (97.3±2.6)% of the cells were derived from the implanted BMSCs in repaired tissue and that the ratio of these cells with proliferative capacity was (76.6±2.5)%. Gross observation suggested most of the cartilage defect areas in Group C were filled with ivory tissue, but those in Group A and B were still obvious depression. Histological staining showed the cartilage defect areas in Group C were filled with cartilage like tissue, which was well integrated with the surrounding normal cartilage, presented a few cartilage lacunas could be seen, and had contents of Col Ⅱ and glycosaminoglycan similar with the adjacent normal cartilage. There was almost no filler in the defect area in Group A. There was little fibrous tissue in the defect area in Group B. ICRS gross score was (8.3±1.0)points in Group C, higher than that in Group A [(0.5±0.6)points] and Group B [(2.3±0.5)points] (P<0.05). ICRS histological score was (10.3±2.4)points in Group C, higher than that in Group A [(0.5±0.6)points] and Group B [(4.5±1.0)points] (P<0.05). Conclusions BMSCs combined with Col Ⅱ-HA-OCS biomimetic hydrogel for repairing porcine articular cartilage defects can achieve satisfactory results. Implanted BMSCs are the main component of the cell composition in the repaired tissue and gradually differentiated into chondrocytes. Key words: Cartilage; Mesenchymal stem cells; Collagen type Ⅱ; Bone regeneration

Preparation of Plasma Membrane Vesicles from Bone Marrow Mesenchymal Stem Cells for Potential Cytoplasm Replacement Therapy.

We have previously reported on the generation of plasma membrane vesicles (PMVs) through the mechanical extrusion of mammalian cells. The fusion of PMVs with mitochondrial deficient Rho0 cells restored mitotic activity under normal culture conditions. Atherosclerosis, type 2 diabetes, Alzheimer's disease, and cancer are age-related diseases that have been reported to be associated with multiple mechanical and functional defects in the cytosol and organelles of a variety of cell types. Bone marrow mesenchymal stem cells (BMSCs) represent a unique cell population from the bone marrow that possess self-renewal capabilities while maintaining their multipotency. The supplementation of senescence cells with young cytoplasm from autologous BMSCs via the fusion of PMVs provides a promising approach to ameliorate or even reverse age-associated phenotypes. This protocol describes how to prepare PMVs from BMSCs via extrusion through a polycarbonate membrane with 3 µm pores, determine the existence of mitochondria and examine the maintenance of membrane potential within PMVs using a confocal microscope, concentrate PMVs by centrifugation, and carry out the in vivo injection of PMVs into the gastrocnemius muscle of mice.

Enhanced critical-sized bone defect repair efficiency by combining deproteinized antler cancellous bone and autologous BMSCs

Previously we have demonstrated that calcinated antler cancellous bone (CACB) has great potential for bone defect repair, due to its highly similar composition and architecture to natural extracellular bone matrix. This study is aiming at seeking for an optimal strategy of combined application of CACB and bone marrow mesenchymal stem cells (BMSCs) in bone defect repair. In vitro study demonstrated that CACB promoted the adhesion, spreading and viability of BMSCs. Increased extracellular matrix production and expression of osteogenic markers in BMSCs were observed when seeded on CACB scaffolds. The cells ceased to proliferation in the dual effect of CACB and osteogenic induction at the early stage of incubation. Hence synergistic effect of CACB combined with autologous undifferentiated BMSCs in rabbit mandible critical-sized defect repair was further evaluated. Histological analysis results showed that loading the CACB with autologous BMSCs resulted in enhanced new bone formation and angiogenesis when compared with implanted CACB alone. These findings indicate that the combination of CACB and autologous BMSCs should become potential routes to improve bone repair efficiency

Low-level laser therapy with helium-neon laser improved viability of osteoporotic bone marrow-derived mesenchymal stem cells from ovariectomy-induced osteoporotic rats.

The purpose of this study was to evaluate the influences of helium–neon (He–Ne) and infrared (IR) lasers on the viability and proliferation rate of healthy and ovariectomy-induced osteoporotic (OVX) bone marrow mesenchymal stem cells (BMMSCs) in vitro. MSCs harvested from the BM of healthy and OVX rats were culture expanded. He–Ne and IR lasers were applied three times at energy densities of 0.6, 1.2, and 2.4??J/cm2 for BMMSCs. BMMSCs viability and proliferation rate were evaluated by MTT assay on days 2, 4, 6, 14, and 21. The results showed that healthy BMMSCs responded optimally to 0.6??J/cm2 using an IR laser after three times of laser radiation. Moreover, it was found that OVX-BMMSCs responded optimally to 0.6??J/cm2 with He–Ne laser and one-time laser radiation. It is concluded that the low-level laser therapy (LLLT) effect depends on the physiological state of the BMMSCs, type of the laser, wavelength, and number of laser sessions. The biostimulation efficiency of LLLT also depends on the delivered energy density. LLLT can enhance the viability and proliferation rate of healthy and especially osteoporotic autologous BMMSCs, which could be very useful in regenerative medicine.

The role of membrane complement regulatory protein in transplantation of bone marrow mesenchymal stem cell

Objective To explore the effect of complement activation on bone marrow mesenchymal stem cells (BMSCs) and evaluate the effect after transfection of complement regulatory proteins. Methods Bone marrow aspirate was harvested from 10 cases of patients suffered from fractures. Mesenchymal stem cells were isolated, indentified cultured and then experimented in vitro. The complement cytotoxicity on the mesenchymal stem cells in autologous serum was measured by Europium cytotoxicity assay. The samples were divided into BMSCs group, BMSCs+ autologous human serum (AHS) group and BMSCs+ inactivated autologous human serum (iAHS) group. The complement membrane attack complex (MAC) deposited on the membranes was detected by flow cytometry. Finally, the cytotoxicity on BMSCs was measured after transfected with membrane complement regulatory proteins (mCRPs). All samples were divided into BMSCs with mCRPs untransfected group and BMSCs with mCRPs transfected group. Results More than 95% of cells derived from bone marrow were identified to be mesenchymal stem cells through detection of cell surface markers by flow cytometry. The cytotoxicity of untreated cells was 0.41%±1.48%. BMSCs harvested from the 10 patients all had cytotoxicity after incubated with autologous serum, and the cytotoxicity was 32.59%±2.73%, while cytotoxicity after incubated with complement inactivated autologous serum was 2.59%±3.08%, which was similar to control group. Complement attack complex (MAC) could be detected on the BMSCs incubated with autologous serum, which implied the complement activation. After transfection of mCRPs, the cytotoxicity of autologous serum on transfected cells was decreased. The cytotoxicity of untransfected cells (41.70%±4.47%) had significant difference compared to the cells transfected with CD55 (21.87%±2.19%), the cells transfected with CD59 (18.67%±1.42%), and the cells transfected with CD46+CD55+CD59 (28.43%±2.14%). CD55, CD59 and CD46+ CD55+CD59 transfected groups could impair effectively the cytotoxicity from complement. However, the cytotoxicity impairment was less effective in CD46 transfected cells (39.30%±3.96%), which had no significant difference compared to untransfected cells. Conclusion Membrane complement regulatory proteins could effectively protect bone marrow mesenchymal stem cells from attacks by complement. Key words: Bone marrow; Mesenchymal stem cells; Complement membrane attack complex

Repair of segmental mandibular bone defects in sheep using bone marrow stromal cells and autologous serum scaffold: a pilot study.

The aim of this study was to evaluate effective bone regeneration using an autologous serum scaffold (alone or seeded with autologous bone marrow-mesenchymal stem cells, BM-MSCs), when implanted in a 30mm length segmental mandibular defect in sheep. The bone defect was filled either with serum scaffold alone (control group; n=5) or combined with BM-MSCs (experimental group; n=10). Bone regeneration was determined at 12 (T12; 2 control sheep and 4 experimental sheep) and 32weeks (T32; 3 control and 6 experimental sheep), as measured by computed and microcomputed tomography and histological examination. Two sheep of the Experimental group died after surgery. While complete bone union in the control group was only observed at T32, it was observed both at T12 (1/4 sheep) and T32 (3/4 sheep) in the experimental group. When properties/characteristics of new bone where compared, a better bone quality, similar to native bone, was observed in the scaffold combined with BM-MSCs. Based on these results, we conclude that the serum scaffold can promote efficient repair of large bone defects, but the combination with BM-MSCs accelerates this process, increasing significantly the amount and quality of bone formed.

Preliminary study on treatment of mandibular osteoradionecrosis by autologous bone marrow mesenchymal stem cells in miniature pigs

To investigate the treatment effect of mandibular osteoradionecrosis (ORN) by autologous bone marrow mesenchymal stem cells (BMMSC) in miniature pigs. Six miniature pigs with mandibular ORN (25 Gy) were used. BMMSC were separated and cultured in vitro and then implanted on to the premolded hydroxyapatite-tricalcium phosphate (HA-TCP). The BMMSC-HA-TCP complexes were implanted into the defective area of mandibular ORN in 4 animals. Implantation of only HA-TCP in 2 animals served as control. Gross observation, spiral CT, coronary CT and histopathologic examination were carried out. Fistula disappeared in animals with BMMSC-HA-TCP complexes 3 months after implantation, while fistula was found in controls. Spiral CT analysis showed that cortical bone repair were found 4 months after BMMSC-HA-TCP complexes implantation, while bone damage (cortical and cancellous bone fracture) increased in controls. Histopathologic examination revealed that 4 months later after BMMSC-HA-TCP complexes implantation, new bone formation and bone cells could be observed, but there was a large number of fibrous tissue and no new bone in controls. Transplantation of autologous BMMSC with HA-TCP may have therapeutic effect in the treatment of mandibular ORN.

Adult autologous mesenchymal stem cells for the treatment of suspected non-infectious inflammatory diseases of the canine central nervous system: safety, feasibility and preliminary clinical findings.

BackgroundNon-infectious inflammatory diseases of the canine central nervous system (CNS) are common idiopathic disorders grouped under the term meningoencephalomyelitis of unknown origin (MUO). Ante mortem diagnosis is achieved via assessment of clinical signs, magnetic resonance imaging (MRI), and cerebrospinal fluid (CSF) analysis, but the definitive diagnosis needs histopathological examination. MUO are mostly considered as autoimmune CNS disorders, so that suppressing the immune reaction is the best management method for patients. Mesenchymal stem cells (MSCs) are under investigation to treat autoimmune and degenerative disorders due to their immunomodulatory and regenerative properties. This study aims to verify the safety, feasibility, and efficacy of MSCs treatment in canine idiopathic autoimmune inflammatory disorders of the CNS.MethodsEight dogs presented with acute onset and rapid progression of multifocal neurological signs were selected to the study. In all patients’ physical and neurological examinations, MRI and CSF analyses were performed. Clinical diagnosis in all cases was MUO. All selected dogs responded initially to immunosuppressive drugs (prednisone and a combination of prednisolone and cytosine arabinoside) but developed undesirable side effects. For all eight dogs, the owners considered euthanasia but accepted cell therapy as a last possibility. Autologous bone marrow MSCs (BMMSCs), isolated, cultured, and expanded, were administered by intrathecal (IT) injection in the cisterna magna intravenously (IV) and by intra-arterial (IA) injection in the right carotid artery. Adverse effects and clinical response were monitored for 6 months up to 2-year follow-up.ResultsThe use of autologous BMMSCs in dogs with MUO was safe for IT, IV, and IA injections. No major short- or long-term adverse effects were registered. All the dogs presented early improvement in their general and neurological conditions, with particular effect on cervical pain. The group of dogs treated by IT+IA administration showed a shorter time of reaction to therapy compared to the group treated by IT+IV administration.ConclusionsMSCs treatment in dogs affected by MOU is safe and feasible. A larger group of dogs is needed to confirm these results as well as CNS histology in order to better understand the underlying mechanisms.

Delayed enrichment for c-kit and inducing cardiac differentiation attenuated protective effects of BMSCs' transplantation in pig model of acute myocardial ischemia.

To investigate the effects of immature cardiomyocytes differentiated from c-kit(+) bone marrow mesenchymal stem cells (BMSCs) against acute myocardial infarction (AMI). Miniswine passage 8 BMSCs were enriched for c-kit and induced by 5 μM 5-azacytidine (AZA) for 14days, and a second enrichment for the dihydropyridine receptor subunit α2δ1 was performed (enriched BMSCs). Thereafter, enriched BMSCs were analyzed by determining cardiac differentiation, secretion function, and the effects of these secreted factors on cardiac stem cells (CSCs). Miniswine with AMI were divided into control, primary BMSCs' (PB), and enriched BMSCs' (EB) groups. Autologous BMSCs were intramyocardially injected into the ischemic regions in PB and EB groups. The following indices were evaluated at different time points, including paracrine of implanted BMSCs, histological and morphological analysis, myocardial perfusion, and cardiac function. As shown by invitro study, enrichment+AZA significantly promoted BMSCs to express cardiac-specific markers and format action potential, but down-regulated the expression of VEGF and bFGF, consequently attenuated BMSCs-inducing CSCs proliferation, migration, and differentiation. The invivo experiments revealed similar results like the invitro 6weeks postoperatively. And in EB group, there were decreased angiogenesis and myocardial perfusion, attenuated resident CSCs-mediated myocardial regeneration, and consequently impaired cardiac function compared with PB group. This pretreatment promoted BMSCs to differentiate into myocardiocytes both invitro and invivo, but impaired their paracrine function and effects on resident CSCs, suggesting that inducing cardiac differentiation alone may not improve protective effects of BMSCs transplantation on AMI.

Bone marrow derived mesenchymal stem cell restore ovarian activity in cases of premature ovarian failure

Introduction: Approximately 1-5% of women worldwide experience cessation of their menstrual cycle prior to age 40, a condition known as POF.The incidence of POF has increased in recent years. As yet, POF cannot be reversed and though some treatments are currently available, improved treatment strategies are urgently required. Regenerative medicine research suggests that stem cells could be used to treat various human diseases, due to their selfrenewal capacity and multiplex differentiation potential. Mesenchymal stem cells (MSCs) are ideal candidates for this purpose. Aim of Work: The purpose of this study is to assess whether bone marrow stem cells (BMSCs) could enter into ovarian niche and differentiate in to oocytes in a mouse model of ovarian failure. Secondly, to explore the therapeutic potency of autologous bone marrow MSC transplantation for primary and chemotherapy-induced ovarian damage and restoration of ovarian function in a pilot human clinical trial. Study Plan: I. Animal study: 18 adult female rats were subjected to cyclophosphamide injection until induction of ovarian failure. 9 mice were injected by 2x106 MSC/Kg . Hormonal and histologic evaluation was done for evidence of regeneration. II. Pilot human trial: 20 cases of premature ovarian failure were enrolled. Autologous bone marrow derived MSC were injected in to the ovaries laparoscopically. Followup was done by measuring serum FSH& follicle counting by ultrasound. Results and Conclusions: All mice of the stem cell treatment group showed positive ovulation with more than 4 ova/low power field. Human subjects showed nomalization of FSH and E2 with sonographic evidence of ovulation in 5 out of 20 patients. These patients were referred to IVF centers.

Recovery of Mesenchymal Stem Cells Homing to Rabbit Myocardial Ischemic Infarct Area by Cu‐microbubble Treatment

Mobilization and homing of bone-marrow mesenchymal stem cells (BMSCs) were observed in acute myocardial ischemic injury, but disappeared in a long-term ischemic stress along with depressed copper (Cu) concentrations in the heart. Cu is required for hypoxia-inducible transcription factor-1 (HIF-1) regulation of expression of BMSC homing factors. The present study was to test the hypothesis that Cu supplementation recovers BMSC homing signaling system, leading to BMSC homing to the chronic ischemic infarct area of heart. Male adult New Zealand rabbits were subjected to coronary artery ligation to generate myocardial ischemia. The autologous BMSCs were labeled with fluorescence and injected via i.v. to the rabbits 24 hours after myocardial ischemia and identified in the ischemic area; but one month after myocardial ischemia, the labeled cells were not found in the ischemic area. A newly developed ultrasound contrast microbubble composed of Cu-albumin surfaced structure was used to specifically deliver Cu to the infarct area, leading to homing of the labeled BMSCs to the chronic ischemic infarct area. AMD3100, the specific SDF-1/CXCR4 axis blocker, blocked the recovery of Cu-microbubble-induced homing of BMSCs. This study thus demonstrated that Cu supplementation reestablishes the signaling pathways for homing of BMSCs to the chronic ischemic area, which involves the role of Cu-activated HIF-1 transcriptional activity. Supported by National Science Foundation of China (81230004 and 81300109).

Randomized trial of autologous bone marrow mesenchymal stem cells transplantation for hepatitis B virus cirrhosis: regulation of Treg/Th17 cells.

Liver cirrhosis is one of the major consequences of hepatitis B virus (HBV) infection, and transplantation of autologous bone marrow mesenchymal stem cells (ABMSCs) is one of promising therapies for patients with HBV-related liver cirrhosis (HBV-LC). However, the mechanism is unclear. The aim of the current study was to explore the role of Treg/Th17 cells in ABMSCs transplantation in patients with HBV-LC. In this prospective study, 56 patients were enrolled and randomly assigned to transplantation group and control group. After 24-week follow-up, 39 patients completed the study (20 cases in transplantation group and 19 cases in control group). The Model for End-Stage Liver Disease scores, liver function, changes of Treg/Th17 cells, as well as related transcription factors and serum cytokines, were determined. Although patients in both groups showed significant improvement after Entecavir treatment, ABMSC transplantation further improved patients' liver function. Moreover, there was a significant increase in Treg cells and a marked decrease in Th17 cells in the transplantation group compared with control, leading to an increased Treg/Th17 ratio. Furthermore, mRNA levels of Treg-related transcription factor (Foxp3) and Th17-related transcription factor (RORγt) were increased and decreased, respectively. In addition, serum transforming growth factor-β levels were significantly higher at early weeks of transplantation, while serum levels of interleukin-17, tumor necrosis factor-α, and interleukin-6 were significantly lower in patients in the transplantation group compared with control. ABMSCs transplantation was effective in improving liver function in patients with HBV-LC, which was mediated, at least in part, through the regulation of Treg/Th17 cell balance.