Autologous Bone Marrow-derived Mesenchymal Stem Research Articles

Autologous bone marrow-derived mesenchymal stem cell transplantation promotes liver regeneration after portal vein embolization in cirrhotic rats

BackgroundPreexisting cirrhosis usually leads to an inadequate and delayed regeneration of the future liver remnant (FLR) after portal vein embolization (PVE). Bone marrow-derived mesenchymal stem cells (BMSC) are promising candidates for therapeutic applications in liver diseases. In this study, the efficacy of autologous BMSCs transplantation to promote FLR regeneration was investigated in a rat cirrhotic model. MethodsAutologous BMSCs were expanded and labeled with PKH26, and then were injected immediately into nonembolized lobes after PVE through portal vein in cirrhotic rat. At 7, 14, and 28 d after this, liver weight and Ki-67 labeling index were measured, and blood analysis was performed. Cirrhotic degree of FLR was assessed by hydroxyproline content assay and histopathology. Gene expressions of vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), interleukin-10 (IL-10), and matrix metalloproteinase-9 (MMP-9) were detected with real-time reverse transcriptase-polymerase chain reaction. Distribution and hepatocyte differentiation of BMSCs in FLR were determined by confocal microscopy. ResultsAutologous BMSCs significantly increased the FLR weight ratio to the total liver and the Ki-67 labeling index, and serum albumin levels were significantly higher and total bilirubin levels were significantly lower in the BMSCs group compared with the controls without BMSCs transplantation 14 and 28 d post-PVE. BMSCs significantly decreased the hydroxyproline content and collagen accumulation, up-regulated the expressions of HGF, IL-10, VEGF, and MMP-9 28 d post-PVE, and expressed hepatocyte-specific markers, such as α-fetoprotein, cytokeratin 18, and albumin in a time-dependent manner in FLR. ConclusionsAutologous BMSCs can differentiate into hepatocyte and promote FLR regeneration after PVE in cirrhotic liver, which may be through improving local microenvironment by decreasing cirrhosis, up-regulating the gene expressions of VEGF, HGF, IL-10, and MMP-9.

A clinical trial report of autologous bone marrow-derived mesenchymal stem cell transplantation in patients with spinal cord injury

Spinal cord injury (SCI) is a severe neurological disease. An effective strategy for the treatment of SCI is urgently required. Stem cell transplantation has emerged as a viable therapeutic option with great potential for restoring neurological function lost following SCI. From 2009 to 2010, a total of 20 SCI patients were enrolled in a clinical trial by Wuhan Hongqiao Brain Hospital; all patients completed and signed informed consent prior to autologous bone marrow-derived mesenchymal stem cell transplantation. Analysis of subsequent treatment results indicated significant improvements in sensory, motor and autonomic nerve function as assessed by the American Spinal Injury Association’s impairment scale. Thirty days after transplantation, a total of 15 patients (75%) demonstrated improvement, including four of the eight patients (50%) with grade A SCI, three of the four patients (75%) with grade B injury and all eight patients (100%) with grade C injury. The most common adverse events, fever and headache, disappeared within 24–48 h without treatment.

Topical Delivery of Cultured Stem Cells to Human Non-Healing Wounds: GMP Facility Development in an Academic Setting and FDA Requirements for an IND and Human Testing

With increasing emphasis on translational research, the need for appropriate regulatory oversight and approval has become essential. The requirements of the Food and Drug Administration (FDA) for Investigational New Drug (IND) exemption in studies that are investigator-initiated have become increasingly stringent. Moreover, academic institutions have not had substantial experience in establishing Good Manufacturing Practice (GMP) facilities required for manipulating human cells in vitro and for chemical or biochemical manufacturing. GMP regulations are established by the FDA under the authority of the Federal Food, Drug and Cosmetic Act. In this report, the authors outline the general strategy and some critical steps that an investigator and the institution may find helpful in developing a GMP facility, especially in an academic center. Also, more specifically and as proof of principle, we describe our approach to culturing autologous bone marrow-derived human mesenchymal stem cells (MSCs) and delivering them to non healing wounds. The lessons learned in this often lengthy and challenging process may be helpful to other academic institutions and investigators embarking on manipulating and delivering viable cells for human experimentation.

A translational approach in using cell sheet fragments of autologous bone marrow-derived mesenchymal stem cells for cellular cardiomyoplasty in a porcine model

Based on a porcine model with surgically created myocardial infarction (MI) as a pre-clinical scheme, this study investigates the clinical translation of cell sheet fragments of autologous mesenchymal stem cells (MSCs) for cellular cardiomyoplasty. MSC sheet fragments retaining endogenous extracellular matrices are fabricated using a thermo-responsive methylcellulose hydrogel system. Echocardiographic observations indicate that transplantation of MSC sheet fragments in infarcted hearts can markedly attenuate the adverse ventricular dilation and preserve the cardiac function post MI, which is in contrast to the controlled groups receiving saline or dissociated MSCs. Additionally, histological analyses suggest that administering MSC sheet fragments significantly prevents the scar expansion and left ventricle remodeling after MI. Immunohistochemistry results demonstrate that the engrafted MSCs can differentiate into endothelial cells and smooth muscle cells, implying that angiogenesis and the subsequent regional perfusion improvement is a promising mechanism for ameliorating post-infarcted cardiac function. However, according to the data recorded by an implantable loop recorder, the transplanted MSCs may provoke arrhythmia. Nevertheless, the proposed approach may potentially lead to the eventual translation of MSC-based therapy into practical and effective clinical treatments.

Proliferation and chondrogenic differentiation of human adipose-derived mesenchymal stem cells in sodium alginate beads with or without hyaluronic acid

Human adipose-derived mesenchymal stem cells (AD-MSCs) have attracted much interest as an alternative to autologous chondrocytes and bone marrow-derived mesenchymal stem cells for cell-based therapy to repair cartilage defects. Sodium alginate (SA) beads have been widely known as a conventional stem cell delivery system for cartilage repair. Hyaluronic acid (HA) has been known to induce cell proliferation and chondrogenic differentiation. Herein, we prepared AD-MSCs-encapsulating SA beads with HA (SA–HA beads) and without HA (SA beads). Then, the morphology, proliferation, and chondrogenic differentiation of AD-MSCs cultured in SA–HA beads or SA beads with a conventional chondrogenic media were evaluated. There was no discernible difference in the morphology of AD-MSCs between SA–HA and SA beads. However, the proliferation (MTT optical density and DNA contents) and chondrogenic differentiation (s-GAG contents and type II collagen staining) of AD-MSCs were significantly enhanced in SA–HA beads as compared to SA beads. The present results suggest that HA can be added to SA beads-based cell delivery systems of AD-MSCs in order to improve their chondrogenesis-inducing capacity for repair of cartilage defects.

The characteristic of minor cell populations in multiple sclerosis patients after cell therapy

Event Abstract Back to Event The characteristic of minor cell populations in multiple sclerosis patients after cell therapy Darya Nizheharodava1*, Alexay Borisov2, Ekatherina Nazarova3, Ekatherina Rudakovskaya1 and Maryna Zafranskaya1 1 Belarussian Medical Academy of Post-Graduated Education, Immunology Department, Belarus 2 Belarusian State Medical University, Neurology Department, Belarus 3 Minsk City Hospital N9, Transplantology Department, Belarus Introduction. Minor cell populations demonstrate functional diversity, from effector action to immunoregulation, depending on pathological conditions. Our previous results showed significant differences in numbers and functions of minor cell populations including γδT-cells, СD8lowT-cells, NK-cells and TNK-cells in multiple sclerosis (MS) patients compared with donors. Objective. To estimate the influence of cell therapy upon minor cell populations’ quantity in peripheral blood of MS patients. Materials and methods. Peripheral blood mononuclear cells (PBMC) were obtained from relapse-remitting MS patients (n=15) with expanded disability status scale (EDSS) score median of 2,5 before and after intravenous infusion of autologous bone marrow-derived mesenchymal stem cells. PBMC were harvested at baseline and during 1 year after cell therapy and monitored with flow cytometer FC500 using appropriate antibodies. Results. γδT-cells, which are able to regulate myelin-specific T-cell response, increased in peripheral blood of MS patients on day 10 after cell therapy (p<0,01) and continued elevation during 1 year of treatment (p<0,05) without any significant changes in СD8+γδT-cells population with potential cytotoxic activity. The percent of СD8lowT-cells reduced after 1 month of cell infusion (p<0,01) and the decrease of СD8lowCD45RO+T-cells and СD8lowCD119+T-cells populations was observed during 9 months after treatment (p<0,01). The last one correlated with activated СD3+CD25+T-cells’ percent (R=0,59, p<0,05). The number of NK-cells correlated with EDSS before treatment (R=0,54, p<0,01) and decreased after 1 month of cell therapy (p<0,05) without significant differences in TNK-cells percentage. Conclusion. Cell therapy established immunoregulatory profile of minor cell populations in peripheral blood of MS patients following 1 year after treatment. Acknowledgements This work was supported by Ministry of Health Care of Republic of Belarus grants N20081618 and N20112544. Keywords: Multiple Sclerosis, cell therapy, minor cell populations, myelin-specific response, Mesenchymal Stem Cells Conference: 15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013. Presentation Type: Abstract Topic: Immune-mediated disease pathogenesis Citation: Nizheharodava D, Borisov A, Nazarova E, Rudakovskaya E and Zafranskaya M (2013). The characteristic of minor cell populations in multiple sclerosis patients after cell therapy. Front. Immunol. Conference Abstract: 15th International Congress of Immunology (ICI). doi: 10.3389/conf.fimmu.2013.02.00590 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 29 May 2013; Published Online: 22 Aug 2013. * Correspondence: Dr. Darya Nizheharodava, Belarussian Medical Academy of Post-Graduated Education, Immunology Department, Minsk, Belarus, nzh@tut.by Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Darya Nizheharodava Alexay Borisov Ekatherina Nazarova Ekatherina Rudakovskaya Maryna Zafranskaya Google Darya Nizheharodava Alexay Borisov Ekatherina Nazarova Ekatherina Rudakovskaya Maryna Zafranskaya Google Scholar Darya Nizheharodava Alexay Borisov Ekatherina Nazarova Ekatherina Rudakovskaya Maryna Zafranskaya PubMed Darya Nizheharodava Alexay Borisov Ekatherina Nazarova Ekatherina Rudakovskaya Maryna Zafranskaya Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Your Cells or Mine?

A new phase I/II study compares autologous and allogeneic mesenchymal stem cells for treating ischemic cardiomyopathy.

Autologous Mesenchymal Stem Cell Therapy in Progressive Multiple Sclerosis: An Open Label Study

Despite updating knowledge and a growing number of medications for multiple sclerosis (MS), no definite treatment is available yet for patients suffering from progressive forms of the disease. Autologous bone marrow derived mesenchymal stem cell (BM-MSC) transplantation is a promising method proposed as a therapy for MS. Although the safety of these cells has been confirmed in hematological, cardiac and inflammatory diseases, its efficacy in MS treatment is still under study. Patients with progressive MS (expanded disability status scale score: 4.0 -6.50) unresponsive to conventional treatments were recruited for this study. Twenty-five patients [f/m: 19/6, mean age: 34.7±7] received a single intrathecal injection of ex-vivo expanded MSCs (mean dose: 29.5×10(6) cells). We observed their therapeutic response for 12 months. Associated short-term adverse events of injection consisted of transient low-grade fever, nausea /vomiting, weakness in the lower limbs and headache. No major delayed adverse effect was reported. 3 patients left the study for personal reasons. The mean (SD) expanded disability status scale (EDSS) score of 22 patients changed from 6.1 (0.6) to 6.3 (0.4). Clinical course of the disease (measured by EDSS) improved in 4, deteriorated in 6 and had no change in 12 patients. In MRI evaluation, 15 patients showed no change, whereas 6 patients showed new T2 or gadolinium enhanced lesions (1 lost to follow-up). It seems that MSC therapy can improve/stabilize the course of the disease in progressive MS in the first year after injection with no serious adverse effects. Repeating the study with a larger sample size, booster injections and longer follow-up using a controlled study design is advised.

A Novel, Minimally-Invasive Technique of Cartilage Repair in the Human Knee Using Arthroscopic Microfracture and Injections of Mesenchymal Stem Cells and Hyaluronic Acid—A Prospective Comparative Study on Safety and Short-Term Efficacy

Most current cell-based cartilage repair techniques require some form of scaffolds and 2 separate surgical procedures. We propose a novel, scaffold-less technique of cartilage repair in the human knee that combines arthroscopic microfracture and outpatient intra-articular injections of autologous bone marrow-derived mesenchymal stem cells (MSCs) and hyaluronic acid (HA). Seventy matched (age, sex, lesion size) knees with symptomatic cartilage defects underwent cartilage repair with the proposed technique (n = 35) or an open technique (n = 35) in which the MSCs were implanted beneath a sutured periosteal patch over the defect. Prospective evaluation of both groups were performed using the International Cartilage Repair Society (ICRS) Cartilage Injury Evaluation Package, which included questions from the Short-Form (SF-36) Health Survey, International Knee Documentation Committee (IKDC) subjective knee evaluation form, Lysholm knee scale, and Tegner activity level scale. Postoperative magnetic resonance imaging (MRI) evaluation was also performed at 1 year for most patients. There were no clinically significant adverse events reported through the course of our study. At the fi nal follow-up (mean = 24.5 months), there was significant improvement in mean IKDC, Lysholm, SF-36 physical component score and visual analogue pain scores in both treatment groups. In the short term, the results of this novel technique are comparable to the open procedure with the added advantages of being minimally invasive and requiring only a single operation under general anaesthesia. Its safety has been validated and its efficacy is currently being evaluated in an ongoing randomised controlled trial.

A Platelet-Rich Plasma-Based Membrane as a Periosteal Substitute with Enhanced Osteogenic and Angiogenic Properties: A New Concept for Bone Repair

The periosteum plays a pivotal role during bone development and repair contributing to bone vascularization and osteoprogenitor cells source. We propose a periosteal substitute engineered using a platelet-rich plasma (PRP) membrane incorporating autologous bone marrow-derived mesenchymal stem cells (PRP/BMSC gel membrane) to be wrapped around an osteoconductive scaffold for regeneration of compromised bone defects. The PRP/BMSC gel membrane was optimized using different compositions for optimal release of vascular endothelial growth factor (VEGF) and platelet derived growth factor-BB (PDGF-BB). Survival and proliferation of cells in the PRP gel membrane with time were confirmed in addition to their osteogenic capacity. Furthermore, to evaluate the possible effects of the PRP/BMSC gel membrane on surrounding progenitor cells in the injury area, we found that the PRP gel membrane products could significantly induce the migration of human endothelial cells in vitro, and increased the expression of bone morphogenetic protein 2 in cultured BMSC. These cells also secreted significant amounts of soluble proangiogenic factors, such as PDGF-BB, VEGF, and interleukin-8 (IL-8). Finally, the functionality of the PRP/BMSC gel membrane periosteal substitute for bone regeneration was tested in vivo both in an ectopic mouse model as well as in a rabbit segmental bone defect model providing evidence of its capacity to biomimic a periosteal response enhancing bone regeneration.

Effect of intracoronary (IC) transplantation of autologous bone marrow-derived mesenchymal stem cells (BMSC) in patients with advanced left ventricular systolic dysfunction

In the absence of cardiac transplantation programs and high cost of CRT-ICD in our community, large numbers of no-option HF patients (pts) for revascularization are still markedly symptomatic. IC-BMSC appears as promising option. 33 (27 males) with chronic HF, LVEF ⩽ 35 and mean age 46 y divided into 2 groups, 17 BMSC pts and 16 pts control. Both were maintained on maximum tolerated medications (follow-up 258 + 47 days). NYHA function class (NYHAFC), six minutes walking test (6MWT), LVESD, LVEDD, 2D-LVEF, systolic (S) and earlydiastolic (E) mitral annulus velocities by TDI evaluated. BMSC were obtained and selectively IC injected. Both groups showed improvement in NYHAFC from 3 to 2, P < 0.001 in BMSC, P < 0.04 for control. 6MWT marginally improved in BMSC from 348 ± 91 to 406 ± 87 meters (m) P < 0.06 while no improvement was noticed in control (361 ± 47 to 336 ± 110M, PNS). LVESD significantly decreased from 6.1 ± 0.8 to 5.7 ± 0.7 cm in BMSC, P < 0.028 while no changes occurred in the control group from 6.7 ± 1.0 to 6.6 ± 1.1 cm, PNS. No changes occurred in LVEDD from 7.1 ± 0.9 to 6.9 ± 0.7 cm in BMSC and from7.6 ± 1.1 to 7.56 ± 1.1 cm in control, PNS for both. No significant changes in LVEF occurred in control (26 ± 6 to 27 ± 7% PNS), while a trend for improvement occurred BMSC from 29 ± 6 to 33 ± 10%, P < 0.059. Mitral S wave showed trend for increase in MBSC (from 4.8 ± 1.5 to 5.3 ± 1.2 cm/s P < 0.058 while no changes occurred in control (5.2 ± 1.7 to 5.2 ± 1.5 cm/s). Mitral E wave showed non-significant increase from 5.7 ± 2.5 to 6.3 ± 2.6 cm/s in BMSC, non-significant decrease was noticed in control from 7.6 ± 2.4 to 7.1 ± 2.7 cm/s, PNS for both. There was no procedure related morbidity or mortality in the BMSC group. On top of standard and maximum tolerated medical treatment for chronic systolic HF, the addition of intracoronary autologous BMSC is feasible, safe and associated with subjective and objective functional improvement with trend towards improvement in parameters of LV performance.

Stem cell therapy in chronic liver disease

To provide an overview of the current status of liver regeneration therapies for liver cirrhosis and future prospects. Various clinical studies for liver disease have been reported, including hepatic administration of autologous CD34-positive cells induced by granulocyte colony-stimulating factor, portal vein administration of CD133-positive mononuclear cells, and administration of autologous bone marrow-derived mesenchymal stem cells. Effectiveness of these approaches has been shown in some patients. We have also reported improved liver fibrosis and function with infusion of autologous bone marrow cells in a basic study with mice, and on the basis of those results started autologous bone marrow cell infusion (ABMi) therapy for liver cirrhosis. The efficacy and safety of ABMi therapy has also been reported by other institutions. Results of recent clinical studies strongly suggest that liver function-improving effects can be achieved using infusion of bone marrow (stem) cells for cirrhosis. New treatment methods using less-invasive bone marrow-derived cultured cells need to be developed.

Transplantation With Autologous Mesenchymal Stem Cells After Acute Myocardial Infarction Evaluated by Magnetic Resonance Imaging

The purpose of this study was to track and investigate the effects of autologous bone marrow-derived mesenchymal stem cells (MSCs) transplantation after acute myocardial infarction in swine assessed by magnetic resonance imaging (MRI). Twenty-four Chinese mini-pigs (27±3 kg) were divided into 4 groups, including control groups (groups 1 and 3) and MSCs transplantation groups (group 2, super paramagnetic iron oxide labeled and group 4, 4',6-diamidino-2-phenylindole labeled). Super paramagnetic iron oxide-labeled and 4',6-diamidino-2-phenylindole-labeled MSCs (3.0×10⁶ cells/mL) with a volume of 10 mL were injected into the left anterior descending artery by a catheter at 1 week after acute myocardial infarction, respectively. Cell distribution, cardiac functions, and scar tissue were quantitatively assessed by MRI. The reduction of the T2* value in the myocardium, spleen, and liver in group 2 was significantly greater than that in group 1. MRI showed that function and scar size at baseline and 3 days after cell infusion were not significantly different between groups 1 and 2. Six weeks later left ventricular ejection fraction (P<0.0001), end-systolic volume (P<0.05), the number of dyskinetic segments (P<0.0001), left ventricular weight index (P<0.0001), and the infarcted size (P<0.0001) in group 4 were all improved comparing with those in group 3. The majority of MSCs entrapped by the extracardial organs were mainly in the spleen. Catheter-based delivery of autologous bone marrow-derived MSCs into infarcted myocardium is feasible and effective.

Transplantation of autologous bone marrow-derived mesenchymal stem cells for traumatic brain injury.

Results from the present study demonstrated that transplantation of autologous bone marrow-derived mesenchymal stem cells into the lesion site in rat brain significantly ameliorated brain tissue pathological changes and brain edema, attenuated glial cell proliferation, and increased brain-derived neurotrophic factor expression. In addition, the number of cells double-labeled for 5-bromodeoxyuridine/glial fibrillary acidic protein and cells expressing nestin increased. Finally, blood vessels were newly generated, and the rats exhibited improved motor and cognitive functions. These results suggested that transplantation of autologous bone marrow-derived mesenchymal stem cells promoted brain remodeling and improved neurological functions following traumatic brain injury.

Investigating the treatment of silicosis with autologous bone marrow-derived mesenchymal stem cells

To explore the safety and curative effects of autologous bone marrow-derived mesenchymal stem cells (BMSCs) in the treatment of silicosis. The protocol was approved by the Ethics Committee of the hospital, and ten patients with silicosis who had given written consent were enrolled in this study. BMSCs isolated from 100 ml of bone marrow for each case were purified and cultured. In each case the 3rd generation of qualified BMSCs (5 × 10(7)) were intravenously administered weekly for 3 weeks. Three cases among 10 patients were treated with BMSCs modified by hepatocyte growth factor (HGF) gene. The clinical symptoms, chest films, chest CT, pulmonary functions, T cells, serum IgG and ceruloplasmin (CP) were observed in 6 or 9 months after treatment. No obvious sub-effect was observed in cases treated with BMSCs, the clinical symptoms (such as cough, sputum and chest tightness) basically disappeared in 9 months after treatment. Pulmonary function tests showed that FVC increased from 71.2% ± 17.0% to 84.0% ± 10.9% (P < 0.01) and FEV1.0 increased from 67.5% ± 17.7% to 80.6% ± 14.9% (P < 0.01). The levels of serum CP and IgG significantly decreased (P < 0.01). Further, the chest films and CT in cases treated with autologous BMSCs modified by HGF gene were improved to different extent. Treatment with autologous BMSCs modified by HGF gene exhibit a beneficial effect on silicosis.

Clinical and laboratory evaluation of patients with end-stage liver cell failure injected with bone marrow-derived hepatocyte-like cells

One of the defining features of the liver is the capacity to maintain a constant size despite injury. Extrahepatic stem cells especially bone marrow-derived stem cells are thought to undertake an important role in liver repopulation. This study was carried out to evaluate the outcome of autologous bone marrow-derived hepatocytes transplantation in patients with end-stage liver cell failure due to chronic hepatitis C. Forty patients were included, divided into two groups. Group I: 20 patients receiving autologous bone marrow-derived mesenchymal stem cells stimulated to hepatic lineage. They were subdivided into two groups regarding the route of transplantation: intrasplenic (10) and intrahepatic (10). Group II: included 20 patients who received traditional supportive treatment. Patients were followed up using examination, laboratory investigations, abdominal ultrasonography, and evaluated by Child score, Model for End Stage Liver Disease score, fatigue scale, and performance status. The results showed significant improvement in group I regarding ascites, lower limb edema, and serum albumin, over the control group. Group I also showed statistically significant improvement in Child score, Model for End Stage Liver Disease score, fatigue scale, and performance status over the controls. No difference was observed between intrahepatic and intrasplenic groups. This study demonstrated the safety and short-term efficacy of autologous bone marrow-derived mesenchymal stem cell injection in liver cell failure. Further study is necessary to standardize the cell dose, determine the life span of the injected cells, and detect the appearance of long-term complications.

Proliferation and chondrogenic differentiation of human adipose-derived mesenchymal stem cells in porous hyaluronic acid scaffold

Human adipose-derived mesenchymal stem cells (AD-MSCs) attracted much interest as a promising alternative to autologous chondrocytes and bone marrow-derived mesenchymal stem cells for cartilage regeneration. Developing a suitable culture technique to direct AD-MSCs into the chondrogenic lineage could be a crucial prerequisite for the cartilage defect repair application of AD-MSCs. Herein, we prepared the PEGDG-crosslinked porous three-dimensional (3D) hyaluronic acid (HA) scaffold and evaluated for its feasibility to induce proliferation and chondrogenic differentiation of the AD-MSCs. In addition, the effect of bone-morphogenetic protein-2 (BMP-2) and platelet-derived growth factor (PDGF) on chondrogenic differentiation was further investigated. Proliferation and chondrogenic differentiation were evaluated by cell morphology, DNA contents, s-GAG contents, and level of mRNA expression of relevant marker genes. When cultured with reference chondrogenic medium (RCM; serum-free DMEM-HG supplemented with 10 ng/mL of transforming growth factor-β1 (TGF-β1), 50 nM ascorbate, 100 nM dexamethasone, and 5 μg/mL of ITS), better proliferation and chondrogenic differentiation of AD-MSCs were obtained in the 3D HA scaffold culture as compared to the micromass culture, a standard 3D culture system. Moreover, the level of chondrogenic differentiation of AD-MSCs in the HA scaffold-RCM culture system was further increased by BMP-2, and decreased by PDGF. These results suggested that the HA scaffold with RCM was a promising chondrogenic culture system of AD-MSCs, and that BMP-2 could potentially serve as a chondrogenic supplement for AD-MSCs. However, PDGF was determined to be an inappropriate supplement based on its inhibition of the chondrogenic differentiation of AD-MSCs.

Topical application of hPDGF-A-modified porcine BMSC and keratinocytes loaded on acellular HAM promotes the healing of combined radiation-wound skin injury in minipigs

Purpose: To evaluate the efficacy of cultured cutaneous substitute (CCS) in accelerating the healing of combined radiation-skin wound injury (CRWI) in minipigs.Material and methods: Autologous porcine bone marrow-derived mesenchymal stem cells (BMSC) and skin-derived keratinocytes (SK) were infected by recombinant retrovirus expressing human (h) platelet-derived growth factor-A (hPDGF-A). CCS was constructed by loading acellular human amniotic membrane (HAM) with normal porcine BMSC and SK (BMSC−/SK−CCS) or with hPDGF-A modified counterparts (BMSC+/SK+CCS). The expression of exogenous hPDGF-A in cells and CCS was assessed by reverse transcription polymerase chain reaction (RT-PCR) and enzyme linked immunosorbent assay (ELISA). The CCS or HAM were grafted to the dorsal CRWI sites (20 Gy local irradiation plus full-thickness skin removal, diameter = 40 mm) of minipigs. Wound healing rate and pathological changes were observed.Results: High levels of hPDGF-A expression were confirmed in gene-modified cells (3780 pg/ml), cultured CCS (506 pg/ml) and transplanted CCS (250 pg/ml). The transplantation of the BMSC+/SK+CCS resulted in a shorter healing time (16–18, days) (P < 0.05 vs. other groups). The healing rates ranked as BMSC+/SK+CCS > BMSC−/SK−CCS > HAM > wound control. Pathologically, there were better granulation formation and re-epithelialisation, and collagen deposition in BMSC+/SK+CCS-treated wound than those in other groups. The angiogenesis ability followed the same order as healing rate of different groups. At day 7, the area densities of vasculature in granulation tissue of group BMSC+/SK+CCS, BMSC−/SK−CCS, HAM, wound only were 15.4, 10.3, 6.0 and 5.7%, respectively, while the number densities of vasculature was 767, 691, 126 and 109 (number/mm2), respectively.Conclusions: Topical transplantation of hPDGF-A modified CCS may be applicable to the management of refractory wounds.

Osteogenic activity of bone marrow-derived mesenchymal stem cells (BMSCs) seeded on irradiated allogenic bone

Allogenic bone grafting, a technique used in orthopaedic surgery, has several problems, including low osteogenic activity. To overcome the problem, this study aimed to determine whether in vivo osteogenesis could be enhanced using allogenic irradiated bone grafts after seeding with autologous bone marrow-derived mesenchymal stem cells (BMSCs). The allogenic bone cylinders were extracted from ACI rats and sterilized by irradiation. Donor BMSCs were obtained from fresh Fischer 344 (F344) rat bone marrow by cell culture. The allogenic bone with or without BMSCs were transplanted subcutaneously into syngeneic F344 rats. At 4 weeks after transplantation, high alkaline phosphatase (ALP) activity, bone-specific osteocalcin mRNA expression and newly formed bone were detected in the allogenic bone with BMSCs. The origin of the newly formed bone was derived from cultured donor BMSCs. However, none of these identifiers of osteogenesis were detected in either the fresh or the irradiated allogenic bone without BMSCs. These results indicate the availability of autologous BMSCs to heighten the osteogenic response of allogenic bone. Our present tissue-engineering method might contribute to a wide variety of allogenic bone grafting techniques in clinical settings.

Safety of autologous bone marrow-derived mesenchymal stem cell transplantation for cartilage repair in 41 patients with 45 joints followed for up to 11 years and 5 months

Among autologous somatic stem cells, bone marrow-derived mesenchymal stem cells (BMSCs) are the most widely used worldwide to repair not only mesenchymal tissues (bone, cartilage) but also many other kinds of tissues, including heart, skin, and liver. Autologous BMSCs are thought to be safe because of the absence of immunological reaction and disease transmission. However, it is possible that they will form tumours during long-term follow-up. In 1988, we transplanted autologous BMSCs to repair articular cartilage, which was the first such trial ever reported. Subsequently we performed this procedure in about 40 patients. Demonstration that neither partial infections nor tumours appeared in these patients provided strong evidence for the safety of autologous BMSC transplantation. Thus, in this study we checked these patients for tumour development and infections. Between January 1998 and November 2008, 41 patients received 45 transplantations. We checked their records until their last visit. We telephoned or mailed the patients who had not visited the clinics recently to establish whether there were any abnormalities in the operated joints. Neither tumours nor infections were observed between 5 and 137 (mean 75) months of follow-up. Autologous BMSC transplantation is a safe procedure and will be widely used around the world.