Injection Of Marrow-derived Mesenchymal Stem Cells Research Articles

Autologous Bone Marrow-Derived Mesenchymal Stem Cells in the Reversal of Unobstructed Azoospermia in Rats.

Non-obstructive azoospermia (NOA) is an important cause of male infertility. This study is being proposed to assess the efficacy of autologous bone marrow-derived mesenchymal stem cells (MSCs) in the reversal of busulfan-induced NOA in rats. Twenty adult 3-month-old male rats were divided into two groups: a control group and a study group. In the study group, bone marrow was aspirated to culture MSCs. NOA was created by stopping endogenous spermatogenesis in all the animals by injecting two doses of busulfan 10 mg/kg body weight with a 3week interval. Four weeks after the last dose of busulfan, two animals were euthanized and the testes were studied histologically to confirm complete azoospermia. In the study group, five million MSCs in 1 mL normal saline were injected into seminiferous tubules; and in the control group, 1 mL of normal saline was injected. After 4weeks of MSC injection, all the rats were euthanized and epididymis tails and testes were harvested and sent for measurement of serological indices, including luminal, cellular, and total diameters, luminal, cellular, and cross-sectional areas, number of tubules per unit area of testis, numerical density of the tubules, and spermatogenesis index, pre- and post-MSC transplantation. The effect of busulfan on the testicular tissue was universally devastating. In the control group, there was variable length and width of markedly necrotic seminiferous tubules, whereas in the group treated with autologous bone marrow-derived MSCs there was variable height of germinal epithelium in seminiferous tubules, with active spermatogenesis, showing spermatogonia, spermatocytes, and sperm. MSC injection in the testis has the potential to reverse the testicular function of spermatogenesis after cytotoxic therapy. Human trials should be undertaken to confirm our findings and bring the results into clinical practice.

Autologous bone marrow derived mesenchymal stem cells are safe for the treatment of Achilles tendinopathy

Achilles tendinopathy is a disabling condition that affects more than 50% of runners. Pre-clinical studies in a large animal model of naturally-occurring tendinopathy similar to human Achilles tendinopathy has shown benefits of autologous bone marrow-derived mesenchymal stem cell (MSC) implantation. However, MSCs are advanced therapies medicinal products (ATMPs), with strict regulatory requirements. Guided by the regulator we carried out a first in man study to assess the safety and efficacy of autologous MSC injection in human patients with non-insertional Achilles tendinopathy. Ten patients, mean age 47 with mid-portion Achilles tendon pain and swelling for more than 6 months, underwent autologous cultured cell injections (median 12.2 × 106, range 5–19 × 106 cells) into their Achilles tendon. At 24 weeks follow-up, no serious adverse reactions or important medical events were observed. MOXFQ, EQ-5D-5L, and VISA-A scores improved clinically at 12 and 24 weeks. VAS pain improved increasingly at 6, 12 and 24 weeks. MOXFQ Pain and VISA-A Scores improved > 12 points from baseline to 24 weeks in 8 patients. Maximum anteroposterior tendon thickness as measured by greyscale US decreased by mean 0.8 mm at 24 weeks. This phase IIa study demonstrated the safety of autologous MSC injection for non-insertional Achilles tendinopathy and provides proof-of-concept of the technique in patients, all of whom had previously failed conservative treatments for chronic disease and leads the way for a larger randomised controlled trial.

Role of bone marrow-derived mesenchymal stem cells in treatment avascular necrosis of femoral head

Avascular necrosis (AVN) of the femoral head (FH) is a debilitating and painful disease with multiple etiologic risk factors head shows an increasing tendency and most commonly affects younger or middle-aged adults. Fifty patients (55 hips) with stage I, II or III avascular necrosis of femoral head were treated by autologous Bone marrow-derived mesenchymal stem cells injection. The patients were followed up clinically and radiologically for a minimum of 2 years at baseline, three, six, 12, and 24 months. The functional outcome was assessed in terms of Harris hip score, and disease progression was assessed radiologically by comparing the preoperative and follow-up MRI at the end of 2 years. On 2-year follow-up, there was considerable improvement in the hip function as measured by the Harris hip score (p = 0.041). On MRI, there was a decrease in the size of the lesion in group A (p = 0.03). Our findings showed that autologous Bone marrow-derived mesenchymal stem cells in avascular necrosis of femoral head is a safe and effective procedure and has better outcome.

Role of bone marrow-derived mesenchymal stem cells in treatment avascular necrosis of femoral head

Avascular necrosis (AVN) of the femoral head (FH) is a debilitating and painful disease with multiple etiologic risk factors head shows an increasing tendency and most commonly affects younger or middle-aged adults. Fifty patients (55 hips) with stage I, II or III avascular necrosis of femoral head were treated by autologous Bone marrow-derived mesenchymal stem cells injection. The patients were followed up clinically and radiologically for a minimum of 2 years at baseline, three, six, 12, and 24 months. The functional outcome was assessed in terms of Harris hip score, and disease progression was assessed radiologically by comparing the preoperative and follow-up MRI at the end of 2 years. On 2-year follow-up, there was considerable improvement in the hip function as measured by the Harris hip score (p = 0.041). On MRI, there was a decrease in the size of the lesion in group A (p = 0.03). Our findings showed that autologous Bone marrow-derived mesenchymal stem cells in avascular necrosis of femoral head is a safe and effective procedure and has better outcome.

Intracerebral Administration of Autologous Mesenchymal Stem Cells as HSV-TK Gene Vehicle for Treatment of Glioblastoma Multiform: Safety and Feasibility Assessment.

Widespread investigation has revealed the promising ability of suicidal genes in the treatment of glioma tumors; nevertheless, promoting their effects relies on the ability to apply suitable vehicles and techniques. In this study, the safety and feasibility of using bone marrow-derived mesenchymal stem cells (MSCs) in combination with prodrug for treatment of patients with primary and secondary glioblastoma multiform (GBM) was assessed. Five GBM patients were recruited. Following gross total resection of the tumor and adjuvant radiotherapy and chemotherapy, intracerebral injection of autologous MSCs transduced with lentivirus containing herpes simplex virus thymidine kinase (HSV-TK) was performed followed by intravenous administration of ganciclovir for 2weeks. The treatment was well tolerated by all patients. Mild-to-moderate fever, headache, and cerebrospinal fluid leukocytosis were evident in three, two, and one patient, respectively. The progression-free survival (PFS) and overall survival (OS) of patients were 95.79 ± 51.40 and 128.85 ± 48.81weeks, respectively. The 1-year PFS and OS were 60% and 100%, respectively, among our patients, and two patients had more than 3years of OS and more than 2years of PFS. It seems that intracerebral administration of bone marrow MSC containing the HSV-TK gene in combination with intravenous ganciclovir would be safe and feasible in the treatment of patients with GBM.

Effects of Bone Marrow and Adipose-Derived Mesenchymal Stem Cells on microRNA Expressions in Acute Alkaline Corneal Burn.

Purpose: The aim of this study was to investigate the microRNA (miRNA) expressions of the corneal tissue after an alkaline burn and to compare the efficiency of adipose- and bone marrow-derived mesenchymal stem cells (MSCs) on expressions. Methods: Thirty-two rats were divided into 4 groups. No intervention was made in the control group. A chemical burn was created by applying 4 μL NaOH soaked in 6 mm filter paper to the right eye of each animal in the other groups. Whereas only subconjunctival 0.1 mL phosphate-buffered saline (PBS) was injected to in the group 1, 2 × 106 adipose- or bone marrow-derived MSC in 0.1 mL PBS was injected subconjunctivally to the animals in the remaining groups (groups 2 and 3, respectively). Tissue samples were collected for miRNA analysis on the third day after the burn. Results: When group 1 was compared with the control group, the expression of 3 of 93 miRNAs increased significantly, whereas the expression of 50 miRNAs decreased significantly. Significant changes in miRNA expressions were observed when group 1 was compared with groups 2 and 3. Although a significant change was observed in the expression of 6 miRNAs in the adipose-derived MSC group, it was found that the expression of 65 miRNAs significantly changed in the bone marrow-derived MSC group. Conclusion: This study shows that there are significant changes in some miRNA expressions after corneal alkaline burn and these changes can be reversed with the subconjunctival injection of MSCs.

Retinal function after intravitreal injection of autologous bone marrow-derived mesenchymal stromal cells in advanced glaucoma

To report electroretinographic (ERG) findings in advanced glaucoma treated with a single intravitreal injection of bone marrow-derived mesenchymal stem cells (MSCs). Intravitreal injection of autologous MSCs (1 × 106 cells) was performed in 2 eyes from 2 patients with open-angle glaucoma in advanced stage of optic neuropathy (ClinicalTrials.gov, NCT02330978, 01.05.2015): cup/disk ratio worse than 0.9, visual field mean deviation index lower than -15dB, visual acuity of light perception, but controlled intraocular pressure. ERG tests were recorded at baseline and week 1, 4 and 48 after injection, using DTL electrodes following the ISCEV standard: After dark adaptation, ERG was elicited using white flashes of 0.01cd.s/m2 and 3.0cd.s/m2, followed by 10-min light adaptation (30cd/m2) and stimuli of 3.0cd.s/m2 and 30Hz flicker. Patients did not show improvement on visual acuity or visual field after treatment. At baseline, ERG responses showed typical findings for advanced glaucoma, with a- and b-wave amplitude and latency within normative range, but reduced photopic negative responses. No noteworthy changes were observed on ERG responses for both cases up to 1week after treatment, but at day 15, one patient showed retinal detachment with proliferative vitreoretinopathy and was removed from the trial. The other patient kept ERG responses stable throughout study period. Although no ERG response changes were observed after MSCs injection in one case, the complication observed on the second one, along with the lack of visual function improvement, warrants further studies involving modified MSCs to treat ocular disorders, including glaucoma. ClinicalTrials.gov, NCT02330978- missed in pdf.

Intravenous transplantation of human muse cells improves blood perfusion in a mouse model of limb ischemia

Abstract Introduction Critical limb ischemia (CLI) is the end-stage of peripheral artery disease caused by atherosclerosis and inflammation. Despite advances in treatment of CLI, substantial number of patients with CLI suffer from severe pain, ulceration, and gangrene. Although the advent of stem cell-based therapy has opened a new avenue for the treatment of various diseases, accumulating evidence suggests current cell-based therapies are safe, but their efficacy is modest in CLI patients. Multilineage-differentiating stress enduring (Muse) cells were first reported by one of the authors and colleagues as endogenous pluripotent-like stem cells residing in the bone marrow, peripheral blood and connective tissue of many organs. Previous studies have demonstrated that systemic administration of exogenous Muse cells improves functional recovery after ischemic organ injury such as myocardial infarction. However, their therapeutic potential in CLI remains unclear. Objective The goal of this study is to elucidate the efficacy and safety of exogenous Muse cell transplantation with animal models of CLI. Methods Muse cells were isolated from human bone marrow-derived mesenchymal stem cells (MSCs) by fluorescence-activated cell sorting with anti-SSEA-3 antibody. To establish an animal model of CLI, 12–14-week-old male BALB/c mice were anesthetized and subjected to left femoral artery and vein resection. At 24 hours after establishment of hindlimb ischemia, mice were randomly divided into 5 groups and treated as follows: Group 1, intravenous injection of PBS as control group; group 2, intravenous injection of 3 × 104 SSEA3-negative MSCs (non-Muse MSCs); group 3, intravenous injection of 3 × 104 Muse cells; group 4, intramuscular injection of 3 × 104 Muse cells into ischemic limb; group 5, intramuscular injection of 2 × 105 mouse bone marrow-derived mononuclear cells (BM-MNCs) into ischemic limb (n=5 for each group). Hindlimb blood flow was evaluated by laser Doppler flowmetry for 14 days and expressed as the ratio of ischemic to non-ischemic hindlimb. Results We found that intramuscular injection of xenogeneic Muse cells without immunosuppression significantly improved the blood flow in the ischemic hindlimb compared to control at day 7 and 14 (0.67 vs. 0.52, p=0.0002 and 0.74 vs. 0.53, p<0.0001, respectively) similar to intramuscular injection of allogenic BM-MNCs. In addition, we also found that intravenous injection of xenogeneic Muse cells without immunosuppression significantly improved the blood flow in the ischemic hindlimb compared to control at day 7 and 14 (0.72, p<0.0001 and 0.67, p=0.004, respectively). In contrast, intravenous injection of xenogeneic non-Muse MSCs did not improve blood flow in the ischemic hindlimb. Conclusion Our result suggests that Muse cell-based regenerative therapy could be a novel, less invasive, cost-effective, and subsequently more effective treatment for CLI patients. Funding Acknowledgement Type of funding source: Public grant(s) – National budget only. Main funding source(s): Grant-in-Aid for Scientific Research (KAKENHI)

Administration of allogeneic mesenchymal stem cells in lengthening phase accelerates early bone consolidation in rat distraction osteogenesis model

BackgroundDistraction osteogenesis (DO) is a surgical technique to promote bone regeneration which may require long duration for bone consolidation. Bone marrow-derived mesenchymal stem cells (MSCs) have been applied to accelerate bone formation in DO. However, the optimal time point for cell therapy in DO remains unknown. This study sought to determine the optimal time point of cell administration to achieve early bone consolidation in DO. We hypothesized that the ratio of circulating MSCs to peripheral mononuclear cells and the level of cytokines in serum might be indicators for cell administration in DO.MethodsUnilateral tibial osteotomy with an external fixator was performed in adult Sprague Dawley rats. Three days after osteotomy, the tibia was lengthened at 0.5 mm/12 h for 5 days. At first, 5 rats were used to analyze the blood components at 6 different time points (3 days before lengthening, on the day lengthening began, or 3, 6, 10, or 14 days after lengthening began) by sorting circulating MSCs and measuring serum levels of stromal cell-derived factor 1 (SDF-1) and interleukin 1β. Then, 40 rats were used for cell therapy study. A single dose of 5 × 105 allogeneic MSCs was locally injected at the lengthening site on day 3, 6, or 10 after lengthening began, or 3 doses of MSCs were injected at the three time points. Sequential X-ray radiographs were taken weekly. Endpoint examinations included micro-computed tomography analysis, mechanical testing, histomorphometry, and histology.ResultsThe number of circulating MSCs and serum level of SDF-1 were significantly increased during lengthening, and then decreased afterwards. Single injection of MSCs during lengthening phase (on day 3, but not day 6 or 10) significantly increased bone volume fraction, mechanical maximum loading, and bone mineralization of the regenerate. Triple injections of MSCs at three time points also significantly increased bone volume and maximum loading of the regenerates.ConclusionThis study demonstrated that bone consolidation could be accelerated by a single injection of MSCs during lengthening when the ratio of peripheral MSCs to mononuclear cells and the serum SDF-1 presented at peak levels concurrently, suggesting that day 3 after lengthening began may be the optimal time point for cell therapy to promote early bone consolidation.

Effect of Mesenchymal Stem Cells Therapy on Cardiovascular Risk in Experimental Diabetic Kidney Disease

Abstract Background Bone Marrow-derived Mesenchymal stem cells (MSC)s were experimentally used in treating diabetes mellitus (DM) and its complications, particularly, diabetic kidney disease (DKD). Cardiovascular diseases are the leading cause of mortality in diabetic patients, which has been strongly associated with progression of DKD. Aim This study was designed to explore the effect of MSCs in modulating cardiovascular risk factors that develop with DKD. Study Design 60 adult female albino rats were allocated into three groups: control group, untreated diabetic kidney disease group (un-DKD) and MSCs treated diabetic kidney disease group (MSC-DKD). DM was induced by a single dose of Streptozotocin at a dose of 35mg/kg, then rats were sacrificed 8 weeks later. MSC-DKD rats were treated by IV injection of bone marrow-derived MSCs 4 weeks after DM induction. DKD establishment was confirmed by histopathological kidney changes and elevated plasma creatinine and urea in both un-DKD and MSC-DKD rats. Methods Blood Pressure and lipid profile were measured, and atherogenic index was calculated to assess cardiovascular risk accompanying the progression of the diabetic renal damage. Moreover, aortic reactivity studies were performed in vitro. Results A single MSCs injection to MSC-DKD rats resulted in significant decrease in final blood glucose, mean arterial blood pressure and atherogenic index, together with a significant increase in plasma HDL level compared to untreated rats, thus minimizing cardiovascular risk factors. MSC-DKD aortae also exhibited significant enhancement of vascular reactivity parameters, particularly vasorelaxation which could partially explain the improvement of blood pressure. On the other hand, DKD didn’t improve by MSCs therapy. These results conformed to tracking labelled MSCs which were found in abundance in both aortic and pancreatic tissues and absent in kidneys. Conclusion MSCs therapy holds hope of improving cardiovascular risk and mortality for diabetic patients with diabetic Kidney disease.

Intraperitoneal Injection of Graphene Oxide Nanoparticle Accelerates Stem Cell Therapy Effects on Acute Kidney Injury.

PurposeGraphene-based nanostructures have shown some degree of stem cell protection against cell death. Acute kidney injury (AKI) is a major cause of mortality in hospitalized patients. Here, graphene oxide (GO) was used to improve the efficacy of bone marrow-derived mesenchymal stem cells (MSCs) in the treatment of AKI induced by cisplatin, a chemotherapy medication used to treat a number of cancers.Materials and MethodsCisplatin-induced AKI was modeled in male rats. Intraperitoneal injection of MSCs mixed with GO, synthesized by graphite powder, H2SO4, and KMnO4 was administered in modeled animals. Biochemical analysis of serum and histological and immunohistochemical (IHC) staining of kidney tissue samples were determined.ResultsAdministration of GO nanoparticles suspended in MSCs reduced serum levels of creatinine (Cr) and blood urea nitrogen (BUN) in cisplatin-induced AKI in the experimental group compared to the control group. Histopathological evaluation also showed an improvement of morphological alterations of kidney, such as cellular proliferation, apoptosis and necrosis, cyst formation and intratubular debris in the experimental group compared to the control group. Our data revealed that GO injection alone without MSCs accelerated the improvement of the kidney injury induced by cisplatin.ConclusionThis study demonstrated that suspended GO could enhance the efficacy of stem cells in the treatment of AKI. GO alone without stem cell accelerates the improvement of cisplatin-induced AKI.

The significant suppressive effect of intravenous injection of autologous bon marrow-derived mesenchymal stem cells on the gene expression and plasma level of CCL5 in patients with refractory rheumatoid arthritis

Background:Rheumatoid arthritis (RA) is the most prevalent autoimmune disease, in which CCL2 and CCL5 are critically involved. The objective was to evaluate the therapeutic effects of bone marrow-derived mesenchymal stem cells (MSCs) on the foregoing chemokines in RA patients.Materials and Methods:Thirteen RA patients were evaluated in terms of clinical manifestations, paraclinical factors, gene expression, and plasma levels of CCL2 and CCL5 prior to treatment and 1 and 6 months after intervention. Real-time-polymerase chain reaction and enzyme-linked immunosorbent assay were employed to assess the gene expression and plasma levels of CCL2 and CCL5 at different time points after MSC therapy. Statistical analysis was performed by SPSS 16 and Prism 7.Results:The CCL2 gene expression had statistically significantly increased (P = 0.034), and its plasma level had insignificantly reduced after 1 month. Furthermore, the gene expression and plasma level of CCL5 had statistically significantly decreased (P = 0.032, P < 0.001). The CCL5 gene expression had statistically significantly increased after 6 months (P = 0.001) and its plasma level had insignificantly reduced.Conclusion:The most significant inhibitory effects of MSC therapy on the gene expression and plasma level of CCL5 were observed at the end of 1 month. The differences between the gene expression and protein levels during the treatment might be related to microRNA effects or the insufficient number of MSC injection.

Transplantation of mesenchymal stem cells causes long-term alleviation of schizophrenia-like behaviour coupled with increased neurogenesis.

Schizophrenia is a neurodevelopmental disease with a mixed genetic and environmental aetiology. Impaired adult hippocampal neurogenesis was suggested both as a pathophysiological mechanism and as a target for therapy. In the present study, we utilized intracerebroventricular transplantation of bone marrow-derived mesenchymal stem cells (MSC) as a means to enhance hippocampal neurogenesis in the ketamine-induced neurodevelopmental murine model for schizophrenia. Syngeneic MSC have successfully engrafted and survived for up to 3 months following transplantation. Improvement in social novelty preference and prepulse inhibition was noted after transplantation. In parallel to behavioural improvement, increased hippocampal neurogenesis as reflected in the numbers of doublecortin expressing neurons in the dentate gyrus and gene expression was noted both 2 weeks following transplantation as well as 3 months later compared with nontreated animals. An independent aging effect was observed for both behaviour and neurogenesis, which was attenuated by MSC treatment. As opposed to MSC treatment, short-term treatment with clozapine was efficient only during treatment and diminished 3 months later. Interestingly, while shortly after transplantation (2 weeks) behavioural improvement was correlated mainly to FGF2 gene expression, 3 months later it was mainly correlated to the expression of the notch ligand DLL1. This suggests that long-term effect during ageing may depend on neural stem cell self-renewal. We conclude that a single intracerebroventricular injection of bone marrow-derived MSC can suffice for long-term reversal of changes in adult hippocampal neurogenesis and improve schizophrenia-like behavioural phenotype inflicted by developmental exposure to ketamine in mice.

Clinical and laboratory outcomes of stem cell transplantation in patients with decompensated liver cirrhosis: single-arm pilot trial

Introduction The high prevalence of hepatitis C virus-associated liver disease has led to increasing number of Egyptian patients having end-stage liver disease and requiring liver transplantation. Because of several limitations of liver transplantation, new alternative treatment modalities are required for patients with liver cirrhosis. Many study results encourage the use of autologous bone marrow-derived mesenchymal stem cells (MSCs) for liver diseases. Aim The aim of this study was to assess the efficacy of using bone marrow-derived MSC on clinical manifestations, liver function, Child–Pugh score, and Model for End-stage Liver Disease score in patients with decompensated liver cirrhosis. Patients and methods This is a pilot single-arm prospective trial that included 13 patients with hepatitis C virus-associated liver cirrhosis who received transdifferentiated MSC via ultrasound-guided percutaneous portal vein infusion plus regular conventional treatment for patients with cirrhosis. Patients were followed up weekly for the first month and monthly for 6 months. Results There was significant improvement in some clinical manifestations, including frequency of attacks of hematemesis, ascites, and lower limb edema after MSC infusion compared with baseline data. Moreover significant increase was detected in mean serum albumin level as it began with 24.62 ± 3.93 and became 27.81 ± 4.14 (P = 0.009) after 6 months of follow-up. Moreover, the results showed improvement in Child–Pugh, but no changes were detected in Model for End-stage Liver Disease score after MSC infusion compared with baseline. Conclusion Our data show that transdifferentiated MSC injection may improve some clinical manifestations and some indices of liver function in patients with decompensated liver cirrhosis with successful tolerability.

Heat shock pretreatment of mesenchymal stem cells for inhibiting\xa0the apoptosis of ovarian granulosa cells enhanced the repair effect on chemotherapy-induced premature ovarian failure

BackgroundPremature ovarian failure (POF) is a severe complication associated with chemotherapy for female patients of childbearing age. A previous study has shown that bone marrow-derived mesenchymal stem cells (MSCs) can partially repair the damaged ovarian structure and function following chemotherapy. Heat shock (HS) is a pretreatment to enhance cell survival. The present study aimed to demonstrate the repair effect and potential working mechanism of HS MSCs on chemotherapy-induced POF.MethodsRat MSCs were isolated, cultured and identified. At 24 h, 48 h and 72 h after different strengths of HS pretreatment for 30 min, 1 h, 2 h and 3 h, apoptosis of MSCs was detected to determine the optimal conditions. Apoptosis and cell proliferation changes of MSCs were detected under the optimal conditions of HS. Apoptosis of HS preconditioned MSCs was detected after adding phosphamide mustard (PM) to mimic the microenvironment under chemotherapy. Rat granulosa cells (GCs) were isolated and cultured. PM was added and apoptosis of GCs was detected after coculture with the pretreated MSCs. The rat model of chemotherapy-induced POF was established and the pretreated MSCs were injected into bilateral ovaries. Ovarian structure and endocrine function were evaluated by ovary weight, follicle count, estrous cycle and sex hormone levels. Apoptosis of GCs was detected by TUNEL assay.ResultsThe apoptosis rate of MSCs with 1 h of HS pretreatment decreased significantly, so 1 h was considered the optimal duration. Under this condition, the reduction in the apoptosis rate persisted until 120 h after the pretreatment and cell proliferation was accelerated. After HS pretreatment, MSCs displayed an increased tolerance to microenvironment under chemotherapy. After coculture with the HS-pretreated MSCs, PM-induced apoptosis of GCs decreased. Injection of the pretreated MSCs into the rat ovaries caused an increase in ovary weight and the number of follicles at different stages of estradiol levels, and a decrease in follicle stimulating hormone levels and apoptosis of GCs in the POF model.ConclusionHS pretreatment enhanced the repair effect of MSCs on chemotherapy-induced POF. The reason for this may be the further vitality enhancement of MSCs, which led to a greater inhibition of apoptosis of GCs.

Treatment Efficiency of Different Routes of Bone Marrow-Derived Mesenchymal Stem Cell Injection in Rat Liver Fibrosis Model

Background/Aims: The most appropriate route for bone marrow-derived mesenchymal stem cell (BM-MSC) transplantation in the management of liver fibrosis remains controversial. This study investigated the therapeutic efficacy of intravenous and intrasplenic BM-MSC transplantation on carbon tetrachloride (CCl4)-induced rat liver fibrosis. Methods: Fifty rats were divided into 5 groups (n = 10 rats per group): healthy control group, CCl4 group, CCl4/ recovery group, CCl4/BM-MSC intravenous group, and CCl4/BM-MSC intrasplenic group. BM-MSCs were isolated, labeled with green fluorescent protein (GFP), and injected into fibrotic rats either intravenously or intrasplenically. Gene expression of interleukins (IL-1β and IL-6), interferon (INF)-γ, hepatic growth factor, and the hepatocyte-specific marker cytokeratin 18 was estimated by quantitative real-time reverse transcription-polymerase chain reaction. Vascular endothelial growth factor and connective tissue growth factor was detected by western blot analysis and enzyme-linked immunosorbent assay, respectively. At 2 weeks after intravenous and intrasplenic BM-MSC injections, GFP-positive cells were detected in liver tissue. Results: Both routes achieved a similar enhancement of liver function, which was confirmed by histopathological examination. The intravenous route was more effective than the intrasplenic route in reducing gene expression levels of IL-1β, IL-6, and INF-γ. However, fibrotic changes were still observed in the recovery group. Conclusion: Intravenous BM-MSC injection was an efficient and appropriate route for BM-MSC transplantation for the management of liver fibrosis.

Study design and rationale for ELPIS: A phase I/IIb randomized pilot study of allogeneic human mesenchymal stem cell injection in patients with hypoplastic left heart syndrome

Despite advances in surgical technique and postoperative care, long-term survival of children born with hypoplastic left heart syndrome (HLHS) remains limited, with cardiac transplantation as the only alternative for patients with failing single ventricle circulations. Maintenance of systemic right ventricular function is crucial for long-term survival, and interventions that improve ventricular function and avoid or defer transplantation in patients with HLHS are urgently needed. We hypothesize that the young myocardium of the HLHS patient is responsive to the biological cues delivered by bone marrow-derived mesenchymal stem cells (MSCs) to improve and preserve right ventricle function. The ELPIS trial (Allogeneic Human MEsenchymal Stem Cell Injection in Patients with Hypoplastic Left Heart Syndrome: An Open Label Pilot Study) is a phase I/IIb trial designed to test whether MSC injection will be both safe and feasible by monitoring the first 10 HLHS patients for new major adverse cardiac events. If our toxicity stopping rule is not activated, we will proceed to the phase IIb component of our study where we will test our efficacy hypothesis that MSC injection improves cardiac function compared with surgery alone. Twenty patients will be enrolled in a randomized phase II trial with a uniform allocation to MSC injection versus standard surgical care (no injection). The 2 trial arms will be compared with respect to improvement of right ventricular function, tricuspid valve annulus size, and regurgitation determined by cardiac magnetic resonance and reduced mortality, morbidity, and need for transplantation. This study will establish the safety and feasibility of allogeneic mesenchymal stem cell injection in HLHS patients and provide important insights in the emerging field of stem cell-based therapy for congenital heart disease patients.

Dual isotope simultaneous imaging to evaluate the effects of intracoronary bone marrow-derived mesenchymal stem cells on perfusion and metabolism in canines with acute myocardial infarction.

Stem cell therapy on acute myocardial infarction (AMI) has been performed for over a decade. In the present study, cardiac perfusion, metabolism and function in dogs with AMI treated by intracoronary injection of bone marrow-derived mesenchymal stem cells (MSCs) were evaluated by dual isotope simultaneous acquisition (DISA) of single positron emission computed tomography (SPECT). Dogs (n=12, 20-30 kg) were randomly assigned to two groups: A graft study (n=6) and control group (n=6). Bone marrow mesenchymal aspirate was collected 3 weeks before surgical procedure. Stem cells were induced by 5-azacytidine for differentiation into myocytes. The dog AMI model was produced by blocking the blood stream at 1/3 of the distinct left anterior descending coronary artery for 90 min. For dogs in the grafting group, MSCs were transplanted by intracoronary injection, and for the control group, 0.9% NaCl was injected instead. At 1 and 10 weeks after MSCs were grafted, respectively, SPECT DISA was performed for each dog in the two groups with 99mTc-SPECT MIBI (925 MBq) and 18F-FDG (222 MBq) for evaluation of myocardial perfusion and metabolism. After the dogs were sacrificed, heart tissue was stained by myocyte-specific antibodies for newborn vessels, troponin T and bromodeoxyuridine (BrdU). Following induction by 5-azacytidine, the morphological features with colony formation, microfilament, as well as atrial granules and positive stainings of α-actinin, myosin and troponin I demonstrated strongly that the MSCs differentiated into myocytes. The number of viable myocardial segments was 10 in the grafting group, which was significantly greater compared with the control group. The ejection fraction of the infarcted left ventricle (LVEF,%) increased from 53.80±9.58 to 70.00±7.52 (change, 16.20±2.93) at 1 and 10 weeks after MSCs engraftment, whilst in the control group, LVEF was 50.50±8.02 and 56.50±7.24 (change, 5.50±2.69), respectively. The LVEF difference was statistically significant (P<0.05) between the graft and control groups. Furthermore, immunostaining of all the myocyte-specific antibodies (for newly born vessels, troponin T and BrdU) was positive. In conclusion, direct intracoronary injection of bone marrow MSCs into injured myocardium in the experimental dog AMI model can significantly improve cardiac function with new vessel formation and myocyte-specific biomarker expression, and in particular, the present study further shows that DISA SPECT can be used for the assessment of stem cell transplantation in the heart.

Long-term influence of bone marrow-derived mesenchymal stem cells on liver ischemia-reperfusion injury in a rat model.

The aim of this study was to evaluate the long-term usefulness of intraportal injection of the bone marrow-derived mesenchymal stem cells (BM-MSCs) in limitation of experimentally induced ischemia-reperfusion injury (IRI) in a rat model. Twenty Wistar rats were divided into 3 groups: donor group (n=5), study group (n=10), and control group (n=5). IRI was performed using a modified hanging-weight system after left portal triad occlusion in study group animals. Isolated autologous BM-MSCs were labeled with fluorochrome PKH-26 then intraportally injected into the rats in the study group. Control group animals were intraportally injected with 1 ml of PBS. Follow-up was 3 months, after which animals were sacrificed for histopathological examination. Migration of BM-MSCs into different organs was examined. H&E staining of liver tissue sections from "time zero" biopsies did not show many irregularities in structural or histological construction compared to liver sections from the control group. However, a small amount of centrilobular hepatocyte necrosis and coagulative necrosis with neutrophil infiltration areas was observed in liver sections of the study group. The migration assay of BM-MSCs labeled with PKH-26 showed the highest positive BM-MSCs staining (6%) in the spleen, while few positively stained cells were found (2%) in liver sections. No BM-MSCs were detected in brain, kidney, or lung tissues. These results suggest that intraportal bone marrow-derived mesenchymal stem cell injection is safe and cells do not migrate chaotically to other organs after targeted implementation.

Bone marrow mesenchymal stem cells enhance bone formation in orthodontically expanded maxillae in rats.

To transplant bone marrow-derived mesenchymal stem cells (MSCs) into the interpremaxillary suture after rapid maxillary expansion with the aim of increasing new bone formation in the suture. Nineteen male Wistar rats were divided into two groups (control, n = 9; experimental, n = 10). Both groups were subjected to expansion for 5 days, and 50 cN of force was applied to the maxillary incisors with a helical spring. Pkh67(+) (green fluorescent dye)-labeled MSCs were applied to the interpremaxillary suture after force application into the interpremaxillary suture of rats. Bone formation in the sutural area was histomorphometrically evaluated, including the amount of new bone formation (µm(2)), number of osteoblasts, number of osteoclasts, and number of vessels. Mann-Whitney U-test was used for statistical evaluation at the P < .05 level. After 10 days of retention, Pkh67(+) can be detected in suture mostly in the injection site under fluorescence microscope. Histomorphometric analysis revealed that a single local injection of MSCs into the midpalatal suture increased the new bone formation in the suture by increasing the number of osteoblasts and new vessel formation, compared with controls injected with phosphate-buffered saline. This preclinical study might provide foundations for the underlying potential clinical use of MSCs after maxillary expansion. Given the fact that MSCs are currently in use in clinical trials, this approach might be a feasible treatment strategy to accelerate new bone tissue formation in midpalatal suture and to shorten the treatment period for patients undergoing maxillary expansion reinforcement.