Allogeneic Bone Marrow Stromal Cells Research Articles

The possibility of using biomedical cell product in the treatment of chronic glomerulonephritis

The basic principles of rational treatment of glomerulonephritis are considered, based on etiology, clinical manifestations and pathomorphological variants of its different forms. Today it has been established that glomerulonephritis can be primary (the etiology is usually unknown) and secondary, when the disease occurs against the background of concomitant pathology (systemic lupus erythematosus, vasculitis, hepatitis, oncological diseases, chronic viral and bacterial infections). The participation of various pathological, genetic and systemic factors in the development of various forms of glomerulonephritis has been shown. The characteristic clinical picture of manifestations of different forms of the disease and the degree of involvement of different renal structures in them are described. Against the background of the severity of the course and prevalence of various forms of glomerulonephritis, it is especially important to develop therapeutic approaches aimed at the full restoration of kidney function and cure of patients. One of such approaches is the use of biomedical cell products based on allogenic mesenchymal multipotent stromal cells and hematopoietic stromal cells. A number of studies have confirmed that the best results can be achieved with the use of cell products in complex therapies with standard treatment methods (use of cytostatics and steroid anti-inflammatory drugs) and alternative drug approaches (combination of monoclonal antibodies and polyenzyme drugs). At the same time, the use of standard and alternative techniques does not lead to a complete recovery of patients, but only transfers the course of the disease from the exacerbation phase to remission. The relevance of further development of biomedical cell products of allogeneic mesenchymal multipotent bone marrow stromal cells is shown, verification of their safety and efficacy in preclinical and clinical studies.

A CK1δ/CK1ε Regulated Metabolic Circuit Is a Therapeutic Vulnerability for Multiple Myeloma

Multiple Myeloma (MM) remains an incurable malignancy, despite the advent of several new therapeutic agents, including immunomodulatory drugs (IMiDs, e.g., Lenalidomide (Len)) and proteasome inhibitors (PIs, e.g., Bortezomib (Btz)). Accordingly, there is an urgent need to identify new targetable vulnerabilities for MM patients. We developed an ex vivo 384-well platform that allows one to define drug sensitivities of primary patient CD138+ MM cells in the context of a reconstructed tumor microenvironment (TME), including allogeneic bone marrow stromal cells, extracellular matrix and MM patient serum. Using this platform and activity-based proteomic profiling (ABPP), we identified shared signaling pathways induced by the interactions of MM with stromal cells and integrated these data with screens performed using a bank of protein kinase inhibitors (PKI) and current anti-MM therapeutics. These analyses revealed that the serine/threonine kinases casein kinase-1δ (CK1δ) and CK1ε as high priority targets for MM. Indeed, a highly selective and potent dual inhibitor of CK1δ/CK1ε coined SR-3029 is the most potent PKI versus MM. Further, our studies revealed SR-3029 has potent activity in 138/153 primary patient MM specimens tested thus far, including quad and penta-refractory MM. Analysis of RNAseq data of over 600 Moffitt Cancer Center (MCC) MM patients revealed that patients with high expression of CK1ε had worse survival outcomes while no survival difference was seen with CK1δ expression. Importantly, using the established 5TGM1/Kal-Ridge (C57B6/KaLwRijHsd) syngeneic mouse model of multiple myeloma, we show that tumors derived from 5TGM1 MM cells, which rapidly die following exposure to SR-3029 ex vivo, are also sensitive to CK1δ/CK1ε inhibition in vivo, where SR-3029 treatment reduced tumor burden and significantly improved survival. Similar results were observed using NSG immune compromised animals inoculated with human MM1.S multiple myeloma cells (both flank and tail vein models), where SR-3029 treated animals had reduced tumor burden and extended survival. Analysis of RNAseq on patients' samples (on stroma) treated ex vivo with SR-3029 revealed CK1δ/CK1ε inhibition suppressed multiple metabolic pathways (oxidative phosphorylation, glycolysis, xenobiotic metabolism). Interestingly, analyses of MCC MM patient RNAseq data revealed upregulation of the genes identified in these metabolic pathways as patients progress from pre-treatment to relapse, and that patient MM samples that were resistant to CK1δ/CK1ε inhibition had an upregulation of some of these metabolic genes. Functional studies are being performed to define the mechanism(s) by which CK1δ/CK1ε inhibition disables MM metabolism. Collectively, these findings establish CK1ε and/or CK1δ as attractive targets for anti-myeloma therapy that are required to sustain MM metabolism. Disclosures Dai: M2Gen: Employment. Shain:Bristol-Myers Squibb: Membership on an entity's Board of Directors or advisory committees; Takeda: Membership on an entity's Board of Directors or advisory committees; Sanofi Genzyme: Membership on an entity's Board of Directors or advisory committees; AbbVie: Research Funding; Janssen: Membership on an entity's Board of Directors or advisory committees; Adaptive Biotechnologies: Consultancy; Amgen: Membership on an entity's Board of Directors or advisory committees; Celgene: Membership on an entity's Board of Directors or advisory committees.

Allogenic chondrocyte/osteoblast-loaded β-tricalcium phosphate bioceramic scaffolds for articular cartilage defect treatment

The medical community has expressed significant interest in the treatment of cartilage defect. Successful repair of articular cartilage defects remains a challenge in clinics. Due to the huge advantages of 3D micro/nanomaterials, 3D artificial micro/nano scaffolds have been widely developed and explored in the tissue repair of articular joints. In this study, chondrocyte/osteoblast-loaded β-tricalcium phosphate (β-TCP) bioceramic scaffold and chondrocyte-loaded β-TCP bioceramic scaffold were prepared by micromass stem cell culture and bioreactor-based cells-loaded scaffold culture for articular cartilage defect treatment. The results demonstrate chondrocyte and osteoblast can be successfully induced from allogeneic bone marrow stromal cells using micromass stem cell culture. Further, chondrocyte-loaded β-TCP scaffold and osteoblast-loaded β-TCP scaffold can be successfully prepared by bioreactor-based cells-loaded scaffold culture. Finally, the scaffolds are applied for Beagle articular cartilage defect treatment. The relative cartilage regeneration abilities on Beagle femoral trochleae were as follows: Chondrocyte/osteoblast-loaded β-TCP bioceramic scaffold group > chondrocyte-loaded β-TCP bioceramic scaffold group > β-TCP bioceramic scaffold. Therefore, micromass stem cell culture and bioreactor-based cells-loaded scaffold culture can be applied to prepare chondrocyte/osteoblast-loaded β-TCP bioceramic scaffold for articular cartilage defect treatment. It suggests allogenic chondrocyte/osteoblast-loaded β-TCP bioceramic scaffold could be potentially used in the treatment of patients with cartilage defects.

Effects of inhalation and intravenous administration of allogeneic mesenchymal bone marrow stromal cells in a bleomycin-induced model of pulmonary fibrosis in rabbits

Aim:to perform a comparative analysis of the effi cacy of the inhaled and intravenous delivery of equivalent doses of bone marrow mesenchymal stem cells (BMMSCs) in rabbits according to the standard model of bleomycin pulmonary fi brosis.Materials and methods.After bronchoscopic instillation of bleomycin, 5 rabbits received intravenous transplantation of 2 × 106 allogeneic BMMSCs, other 5 rabbits – 2 × 107 MSCs inhaled via compressor nebulizer; control healthy and bleomycin group included 5 animals each.Results.Both groups treated with BMMSCs had a signifi cantly lower Ashcroft fi brosis index than the bleomycin control group. Expression of collagen in lung tissue in all groups with bleomycin injury was superior to healthy controls, but in animals underwent intravenous BMMSC transplantation collagen score was 0.74 points, and in inhaled treated group – 0.51 points, while in bleomycin controls – 2.1 point. Levels of TNF-α and TGF-β1 in BAL fl uids tended to decrease in treatment groups, but did not differ signifi cantly from control. A similar picture was observed in the cytological analysis of BAL.Conclusion.In general, both methods of delivering of BMMSCs to the lungs demonstrated similar therapeutic effects in inhibiting the development of experimental fi brosis, indicating that both intravenous and inhalational way of introduction can be used for subsequent clinical studies.

Experimental therapy of graft-versus-host disease by mesenchymal stromal cells grown on oxide nanocoatings

Immune aggression to transplanted allogeneic bone marrow, i.e. the graft-versus-host disease (GVHD), could be decreased by the suppression of effector and/or activation of T- regulatory cells (Treg). This task could be solved by co-transplantaiton of allogeneic bone marrow and mesenchymal stromal cells (MSCs). This study demonstrated the elevated immune modulating activity of MSCs by their culturing in vitro on Al₂0₃ oxide nanocoatings. Introduction of the cells to the animals with GVHD resulted in an increased content of Treg in the spleen of bone marrow recipients, reduced severity of the pathology, and higher survival of animals. Thefindings could be the basis for developing the new approaches to optimize the GVHD treatment methods involving the oxide nanocoating cultured MSCs.

Experimental bladder regeneration using a poly-l-lactide/silk fibroin scaffold seeded with nanoparticle-labeled allogenic bone marrow stromal cells.

In the present study, a poly-l-lactide/silk fibroin (PL-SF) bilayer scaffold seeded with allogenic bone marrow stromal cells (BMSCs) was investigated as a potential approach for bladder tissue engineering in a model of partial bladder wall cystectomy in rabbits. The inner porous layer of the scaffold produced from silk fibroin was designed to promote cell proliferation and the outer layer produced from poly-l-lactic acid to serve as a waterproof barrier. To compare the feasibility and efficacy of BMSC application in the reconstruction of bladder defects, 12 adult male rabbits were divided into experimental and control groups (six animals each) that received a scaffold seeded with BMSCs or an acellular one, respectively. For BMSC tracking in the graft in in vivo studies using magnetic resonance imaging, cells were labeled with superparamagnetic iron oxide nanoparticles. In vitro studies demonstrated high intracellular incorporation of nanoparticles and the absence of a toxic influence on BMSC viability and proliferation. Following implantation of the graft with BMSCs into the bladder, we observed integration of the scaffold with surrounding bladder tissues (as detected by magnetic resonance imaging). During the follow-up period of 12 weeks, labeled BMSCs resided in the implanted scaffold. The functional activity of the reconstructed bladder was confirmed by electromyography. Subsequent histological assay demonstrated enhanced biointegrative properties of the PL-SF scaffold with cells in comparison to the control graft, as related to complete regeneration of the smooth muscle and urothelium tissues in the implant. Confocal microscopy studies confirmed the presence of the superparamagnetic iron oxide nanoparticle-labeled BMSCs in newly formed bladder layers, thus indicating the role of stem cells in bladder regeneration. The results of this study demonstrate that application of a PL-SF scaffold seeded with allogenic BMSCs can enhance biointegration of the graft in vivo and support bladder tissue regeneration and function.

Bone Marrow Stromal Cell Intraspinal Transplants Fail to Improve Motor Outcomes in a Severe Model of Spinal Cord Injury.

Bone marrow stromal cells (BMSCs) have been reported to exert potential neuroprotective properties in models of neurotrauma, although precise mechanisms underlying their benefits are poorly understood. Despite this lack of knowledge, several clinical trials have been initiated using these cells. To determine whether local mechanisms mediate BMSC neuroprotective actions, we grafted allogeneic BMSCs to sites of severe, compressive spinal cord injury (SCI) in Sprague-Dawley rats. Cells were administered 48 h after the original injury. Additional animals received allogeneic MSCs that were genetically modified to secrete brain-derived neurotrophic factor (BDNF) to further determine whether a locally administered neurotrophic factor provides or extends neuroprotection. When assessed 2 months post-injury in a clinically relevant model of severe SCI, BMSC grafts with or without BDNF secretion failed to improve motor outcomes. Thus, allogeneic grafts of BMSCs do not appear to act through local mechanisms, and future clinical trials that acutely deliver BMSCs to actual sites of injury within days are unlikely to be beneficial. Additional studies should address whether systemic administration of BMSCs alter outcomes from neurotrauma.

Local delivery of allogeneic bone marrow and adipose tissue-derived mesenchymal stromal cells for cutaneous wound healing in a porcine model

Wound healing remains a major challenge in modern medicine. Bone marrow- (BM) and adipose tissue- (AT) derived mesenchymal stromal/stem cells (MSCs) are of great interest for tissue reconstruction due to their unique immunological properties and regenerative potential. The purpose of this study was to characterize BM and AT-MSCs and evaluate their effect when administered in a porcine wound model. MSCs were derived from male Göttingen Minipigs and characterized according to established criteria. Allogeneic BM- or AT-MSCs were administered intradermally (1 x 10(6) cells) into partial-thickness wounds created on female animals, and covered with Vaseline® gauze or fibrin in a randomized pattern. Animals were euthanized at 7, 10, 14 and 21 days. Tissues were analyzed visually for healing and by microscopic examination for epidermal development and remodelling. Polymerase chain reaction (PCR) was used to detect the presence of male DNA in the specimens. All wounds were healed by 14 days. MSC-injected wounds were associated with improved appearance and faster re-epithelialization compared to saline controls. Evaluation of rete ridge depth and architecture showed that MSC treatment promoted a faster rate of epidermal maturation. Male DNA was detected in all samples at days 7 and 10, suggesting the presence of MSCs. We showed the safety, feasibility and potential efficacy of local injection of allogeneic BM- and AT-MSCs for treatment of wounds in a preclinical model. Our data in this large animal model support the potential use of BM- and AT-MSC for treatment of cutaneous wounds through modulation of healing and epithelialization.

P352 The effectiveness and safeness of allogeneic mesenchymal bone marrow stromal cells in patients with refractory Crohn's disease – 5 years of observation

P352 The effectiveness and safeness of allogeneic mesenchymal bone marrow stromal cells in patients with refractory Crohn's disease – 5 years of observation

Cell therapy with bone marrow stromal cells after intracerebral hemorrhage: impact of platelet-rich plasma scaffolds

Cell therapy using bone marrow stromal cells (BMSCs) has been considered a promising strategy for neurologic sequelae after intracerebral hemorrhage (ICH). However, after intracerebral administration of BMSCs, most of the cells die, partly because of the absence of extracellular matrix. Intracerebral transplantation of BMSCs, supported in a platelet-rich plasma (PRP) scaffold, optimizes this type of cell therapy. ICH was induced by stereotactic injection of 0.5 IU of collagenase type IV in the striatum of adult Wistar rats (n = 40). Two months later, the rats were subjected to intracerebral administration of 5 × 10(6) allogeneic BMSCs embedded in a PRP scaffold (n = 10), 5 × 10(6) allogeneic BMSCs in saline (n = 10), PRP-derived scaffold only (n = 10) or saline only (n = 10). Functional improvements in each group over the next 6 months were assessed using Rotarod and Video-Tracking-Box tests. Endogenous neurogenesis and survival of transplanted BMSCs were examined at the end of follow-up. Our study demonstrated neurologic improvement after BMSC transplantation and significantly better functional improvement for the group of animals that received BMSCs in the PRP-derived scaffold compared with the group that received BMSCs in saline. Histologic results showed that better functional outcome was associated with strong activation of endogenous neurogenesis. After intracerebral administration of BMSCs, donor cells were integrated in the injured tissue and showed phenotypic expression of glial fibrillary acidic protein and neuronal nucleus. PRP-derived scaffolds increase the viability and biologic activity of BMSCs and optimize functional recovery when this type of cell therapy is applied after ICH.

Allogeneic bone marrow stromal cell transplantation after cerebral hemorrhage achieves cell transdifferentiation and modulates endogenous neurogenesis

When a severe neurologic lesion occurs as a consequence of intracerebral hemorrhage (ICH), there is no effective treatment available for improving the outcome. However, cell therapy has opened new perspectives on reducing neurologic sequels subsequent to this disease. In this study, ICH was induced by stereotactic injection of 0.5 U collagenase type IV in the striatum of adult Wistar rats, and 2 h later a group of animals (n = 48) was subjected to intracerebral injection of 2 × 10(6) allogeneic bone marrow stromal cells (BMSC), while a control group (n = 48) received saline only. Eight animals from each group were killed at 48 h, 72 h, 7 days, 14 days, 21 days and 28 days. At these time-points, endogenous neurogenesis and survival of transplanted BMSC were studied. Our findings show that after allogeneic BMSC transplantation, donor cells can survive in the brain tissue expressing neuronal and astroglial markers. Furthermore, BMSC transplantation enhances endogenous neurogenesis and inhibits apoptosis of newborn neural cells. Although these results should be extrapolated to human disease with caution, it is obvious that cell therapy using allogeneic BMSC transplantation offers great promise for developing novel and efficacious strategies in patients suffering ICH.

Exogenous VEGF-C Augments the Efficacy of Therapeutic Lymphangiogenesis Induced by Allogenic Bone Marrow Stromal Cells in a Rabbit Model of Limb Secondary Lymphedema

To determine the effect of bone marrow stromal cells transplantation and vascular endothelial growth factor C administration as a treatment for secondary lymphedema. Bone marrow stromal cells and/or vascular endothelial growth factor C protein were injected into a rabbit model of limb lymphedema. Water displacement volumetry was performed to measure limb volume changes. Immunohistochemistry was performed to detect VEGFR-3 and to count lymph vessel. Western blot analysis was performed to detect vascular endothelial growth factor C. Before treatment, rabbits had an average volume of edema in the limb of 61.25 ± 5.28, 62.37 ± 4.97, 60.58 ± 7.18 and 61.79 ± 4.33 ml (P = 0.753), respectively, in the BMSC + VEGF-C, bone marrow stromal cell, vascular endothelial growth factor C and control groups. With therapy, this was reduced to an average volume of 7.60 ± 3.02, 12.78 ± 3.41, 31.55 ± 3.51 and 62.33 ± 6.59 ml, respectively, in the four groups 6 months after treatment. Quantitative analysis showed that the vessel numbers were significantly increased in the BMSC + VEGF-C, bone marrow stromal cell and vascular endothelial growth factor C groups compared with the control group at 28 days after the operation (P< 0.05). Western blot analysis demonstrated that expression of vascular endothelial growth factor C was higher in the BMSC + VEGF-C and BMSC groups. The combined treatment with bone marrow stromal cell transplantation and vascular endothelial growth factor C administration is superior to bone marrow stromal cell transplantation alone in the treatment of limb lymphedema. Bone marrow stromal cell transplantation and vascular endothelial growth factor C administration could enhance the therapeutic effect of each other.

Late transplantation of allogeneic bone marrow stromal cells improves neurologic deficits subsequent to intracerebral hemorrhage

Stem cell therapy seems to be a promising therapeutic tool for treating central nervous system (CNS) injuries. Bone marrow stromal cell (BMSC) transplantation influences functional outcome subsequent to intracerebral hemorrhage (ICH), and enhances endogenous neurogenesis in acute condition studies. We investigated whether late administration of BMSC improves functional deficits subsequent to ICH. Experimental ICH was induced by stereotactic injection of 0.5 IU collagenase type IV in the striatum of adult female Wistar rats, and 2 months later intralesional administration of 5 × 10(6) allogeneic BMSC from male donors rats in saline (n = 10), or saline only (n = 10), was performed. In the following 6 months, functional outcome was evaluated in each animal by rotarod, modified neurologic severity score (mNSS) and video-tracking box (VTB) tests. To study the behavior of BMSC after transplantation, in situ hybridization studies were performed, with double labeling of the chromosome Y-linked SrY-gene, and neuronal nuclei (NeuN) protein or gliofibrillary acidic protein (GFAP). The assessment test revealed significant improvements in functional outcome for the BMSC-treated animals after 2 months of follow-up. Histologic results showed that functional outcome was associated with strong reactivation of endogenous neurogenesis. Furthermore, intralesional BMSC not only integrated in the injured tissue but also showed phenotypic expression of GFAP and NeuN. Late intracerebral transplantation of allogeneic BMSC induces functional recovery after ICH. The possibility of using this type of cell therapy to reverse the consequences of hemorrhagic stroke in humans should be considered.

SCID-Synth-Hu: a Novel Multiple Myeloma Model for In Vivo Expansion of Primary Cells

AbstractAbstract 452The critical role of the human bone marrow microenvironment (HuBMM) in the pathogenesis of Multiple Myeloma (MM) has recently allowed the design of novel therapeutical approaches targeting not only MM cells but also their specific HuBMM. However, the lack of adequate mouse models, capable to recapitulate a HuBMM, has restrained large scale in vivo screening of investigational drugs. In fact, only the SCID-hu model, in which human MM cells are grown in human fetal bone chips previously implanted in SCID mice, addresses this specific requirement. However the poor availability of human fetal bone chips, the allogeneic nature of the fetal BM milieu versus MM cells and the heterogeneity of implanted human bone chips are important restrains of this system. Here we report the development of a novel in vivo model of human MM (SCID-synth-hu), which is based on the implant of a three-dimensional (3-D) poly-ε-caprolactone polymeric scaffold (PCLS) into SCID mice as recipient to allow growth of MM cells in a reconstituted HuBMM. This biosynthetic scaffold has been designed to resemble the micro-architecture of a normal human adult bone and was characterized by 3-D interconnected large and small pores suitable for engraftment of bone marrow-derived cells. Human bone marrow stromal cells (BMSCs) were collected from BM aspirates from MM patients and firstly used for coating 3D internal surface of PCLSs. We performed in vitro dynamic seeding of BMSCs into PCLSs using a suspension of 106 cells in 500 μl of growth medium. Before implantation, PCLSs were incubated in complete medium at 37°C, in 5% CO2 for 24 hours, in order to allow cell adhesion on 3D surfaces. Then, PCLSs were implanted subcutaneously into SCID mice. CD138+ immune-selected primary MM cells, obtained by MM patient with a different disease status, were directly injected into PCLSs, which have been previously coated with allogeneic BMSCs 2–3 weeks after the in vivo implant. By this experimental approach, we achieved engraftment of primary MM cells in PCLSs within a non autologous bone marrow milieu. In a subsequent series of experiments, bone marrow mononuclear cells (BMMCs), obtained by Ficoll gradient separation and containing primary unselected CD138+ and their autologous BMSCs, were directly seeded in vitro into PCLSs which were implanted in SCID mice after 24 hours of incubation. At different time points, H&E and CD138 or κ/λ staining demonstrated engraftment and filling of 3-D spaces by primary MM cells within the autologous bone marrow milieu in PCLSs retrieved from SCID-synth-hu mice. Neo-synthesized extracellular matrix and angiogenesis were also shown by H&E and immune histochemical staining in retrieved PCLSs. Angiogenesis mostly occurred within areas of MM infiltration, suggesting its role in our system. In vivo MM growth was monitored by ELISA measuring of human monotypic immunoglobulins (Igs) in mouse sera 4 to 10 weeks after cell injection. To demonstrate the usefulness of our SCID-synth-hu model as an experimental platform for in vivo testing of investigational drugs, mice bearing human MM implants were treated intraperitoneally with bortezomib plus dexamethasone (Bort+Dex). As expected, SCID-synth-hu mice treated with Bort+Dex exhibited a significant decrease of monotypic light chains in mice sera and induction of apoptosis of MM cells in retrieved PCLSs, as compared to untreated control mice. Our experimental findings demonstrate that the SCID-synth-hu is the first experimental system which allows the in vivo expansion of human primary MM cells within their autologous adult HuBMM. The unlimited availability and the low cost of PCLSs, as well as the potential for dissecting the biological events within the HuBMM, represent a clear improvement over the available preclinical in vivo models of MM. We conclude that the SCID-synth-hu is a unique tool for large scale in vivo preclinical screening of novel agents targeting MM in its autologous HuBMM, and a novel resource for translational research in the experimental treatment of this still incurable disease. Disclosures:No relevant conflicts of interest to declare.

Repair of Bone Defect in Caprine Tibia Using a Laminated Scaffold With Bone Marrow Stromal Cells Loaded Poly (L-Lactic Acid)/β-Tricalcium Phosphate

Repair of bone defects of a critical size encounters many problems, and many efforts aim to build a porous scaffold loading bone marrow stromal cells (BMSCs) or bone morphogenetic protein (BMP2) to quickly repair bone defects. In this paper, a laminated scaffold was designed and tested for the repair of bone defects in a caprine tibia. Beta-tricalcium phosphate (β-TCP) and poly (L-lactic acid) (PLLA) were fabricated to a sandwich structured composite that was then rolled up to form a cylindrical shaped, porous scaffold. The porosity and bending strength of the PLLA/β-TCP laminated scaffold were around 70% and 1.7 MPa, respectively. Results from in vitro experiments showed that the pH value of the scaffold in water fluctuated between 4.9 and 7.0 during its degradation. When exposed to the simulated body fluid, the scaffold lost its strength after 11 weeks of degradation. After implantation in Chinese caprines' diaphyseal defects with loaded allogeneic BMSCs, the scaffold sped up the bone repair without collapse of the scaffold and the unwanted inflammatory response, and then rapidly degraded and finally disappeared at 12 weeks. Gross examinations and pullout tests showed that the experimental caprines walked normally and the implanted leg could be heavily loaded. X-ray examinations and histological analyses showed new bone tissues formed with similar structures to normal ones. It is suggested that the novel PLLA/β-TCP laminated scaffold with BMSCs loading can regenerate new bones quickly.

Effect of self-assembling peptide, chondrogenic factors, and bone marrow-derived stromal cells on osteochondral repair

The goal of this study was to test the ability of an injectable self-assembling peptide (KLD) hydrogel with or without chondrogenic factors (CF) and allogeneic bone marrow stromal cells (BMSCs) to stimulate cartilage regeneration in a full-thickness, critically-sized, rabbit cartilage defect model in vivo. We used CF treatments to test the hypotheses that CF would stimulate chondrogenesis and matrix production by cells migrating into acellular KLD (KLD+CF) or by BMSCs delivered in KLD (KLD+CF+BMSCs). Three groups were tested against contralateral untreated controls: KLD, KLD+CF, and KLD+CF+BMSCs, n=6-7. Transforming growth factor-β1 (TGF-β1), dexamethasone, and insulin-like growth factor-1 (IGF-1) were used as CF pre-mixed with KLD and BMSCs before injection. Evaluations included gross, histological, immunohistochemical and radiographic analyses. KLD without CF or BMSCs showed the greatest repair after 12 weeks with significantly higher Safranin-O, collagen II immunostaining, and cumulative histology scores than untreated contralateral controls. KLD+CF resulted in significantly higher aggrecan immunostaining than untreated contralateral controls. Including allogeneic BMSCs+CF markedly reduced the quality of repair and increased osteophyte formation compared to KLD-alone. These data show that KLD can fill full-thickness osteochondral defects in situ and improve cartilage repair as shown by Safranin-O, collagen II immunostaining, and cumulative histology. In this small animal model, the full-thickness critically-sized defect provided access to the marrow, similar in concept to abrasion arthroplasty or spongialization in large animal models, and suggests that combining KLD with these techniques may improve current practice.

Safety and Efficiency of Transplantation of Allogenic Multipotent Stromal Cells in Surgical Treatment of Dilatated Cardiomyopathy

Clinical study of intracoronary transplantation of allogenic multipotent bone marrow stromal cells was carried out in patients with severe chronic cardiac failure against the background of dilatated cardiomyopathy. The results indicate that intracoronary injection of allogenic multipotent stromal precursors is a safe procedure. No complications and side effects directly or indirectly related to cell transplantation were recorded during the immediate and delayed postoperative periods. The positive effect of cell transplantation developed from week 1 after transplantation and persisted for 6 months. It manifested in reduction of the level of brain natriuretic peptide and improvement of patient's functional status and quality of life. No appreciable changes in the main echocardiographic values were noted. Transplantation of allogenic multipotent stromal cells is effective as a component of combined therapy for chronic cardiac failure at the stage of preparation to surgery as a "bridge to surgical treatment".

Intralesional administration of allogeneic bone marrow stromal cells reduces functional deficits after intracerebral hemorrhage

When a severe neurological lesion occurs as a consequence of intracerebral bleeding, no effective treatment for improving the outcome is currently available. In the present study, intracerebral hemorrhage (ICH) was induced by stereotactic injection of 0.5 U of collagenase type IV in the striatum of adult Wistar rats, and three days later, intralesional administration of 2 x 10(6) allogeneic bone marrow stromal cells (BMSC) in saline (n:10), or saline only (n:10), was performed. In the following 30 days, functional outcome was evaluated in each animal by rotarod and the modified neurological severity score (mNSS) test. Progressive and functional improvement was observed in BMSC-transplanted rats compared with controls, together with morphological images suggesting that intracerebral administration of BMSC increases endogenous neurogenesis at the level of subventricular zone (SVZ). These findings suggest that local administration of allogeneic BMSC could be useful to reduce the neurological deficits caused by intracerebral hemorrhage.

Reducing macrophages to improve bone marrow stromal cell survival in the contused spinal cord

We tested whether reducing macrophage infiltration would improve the survival of allogeneic bone marrow stromal cells (BMSC) transplanted in the contused adult rat thoracic spinal cord. Treatment with cyclosporine, minocycline, or methylprednisolone all resulted in a significant decrease in macrophage infiltration at 3 days postinjury. However, when BMSC were injected at that time point, survival 7 days later was similar between treatment groups and saline-injected controls. In fact, we found that the presence of BMSC resulted in a significant increase in macrophage infiltration into the contusion.

What's New in Cardiac Cell Therapy? Allogeneic Bone Marrow Stromal Cells as “Universal Donor Cells”

Cardiac cell therapies offer distinct and exciting advantages over current treatments to prevent postinfarction heart failure because they can reverse ventricular remodeling and improve function, but only if the implanted stem cells contribute biological functions and achieve prolonged engraftment within the hostile environment of the damaged heart. Unfortunately, function is diminished in autologous stem cells isolated from older patients and those with comorbidities, and so clinical trials testing the implantation of healthy, allogeneic bone marrow-derived stromal cells (MSCs) isolated from young donors are currently underway. MSCs are unique because, in addition to exerting paracrine effects that restore blood flow and recruit endogenous stem cells to the infarct, they exhibit immune-modulating properties in culture that-if retained after allogeneic implantation-imply the cells may escape immune recognition within the heart. At present, the scope of MSC immune modulation after implantation is unclear.