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| Carrier-free epithelial cell sheets prepared by enzymatic degradation of collagen gel.
July 7, 2010 at 12:59 PM
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| | Carrier-free epithelial cell sheets prepared by enzymatic degradation of collagen gel. J Tissue Eng Regen Med. 2010 Jul 5; Authors: Ke Q, Wang X, Gao Q, Wu Z, Wan P, Zhan W, Ge J, Wang Z Limbal stem-cell deficiency by ocular trauma or disease causes corneal opacification and vision loss. Conventional tissue engineering using biodegradable scaffolds has met with limited success. In this study, we developed a novel method for preparing carrier-free epithelial cell sheets, which have potential for use in repairing defects of the ocular surface. Stratified corneal epithelial cell sheets were prepared in culture dishes coated with biodegradable type I collagen. Haematoxylin and eosin staining, electron microscopy and immunohistochemistry were performed to characterize the cell sheets. Then, carrier-free epithelial sheets were successfully engineered using specific collagenase to degrade the collagen gel. Cell sheets of four to six cell layers after culture for 14 days were similar to natural rabbit corneal epithelia, as shown by pathological examination. Microvillus, tight cell-cell junctions and desmosome junctions were observed via electron microscopy. K3 and basement membrane components, such as type IV collagen and laminin, were expressed in the cells sheets and integrin beta1 was maintained in basal cells. This novel method of using collagenase to degrade collagen gel is both simple and effective in preparing intact carrier-free epithelial cell sheets. Such sheets have great potential for application during in vivo corneal regeneration. Copyright (c) 2010 John Wiley & Sons, Ltd. PMID: 20603893 [PubMed - as supplied by publisher] | | |
| Interaction between electrical stimulation, protein coating and matrix elasticity: a complex effect on muscle fibre maturation.
July 7, 2010 at 12:59 PM
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| | Interaction between electrical stimulation, protein coating and matrix elasticity: a complex effect on muscle fibre maturation. J Tissue Eng Regen Med. 2010 Jul 5; Authors: Boonen KJ, van der Schaft DW, Baaijens FP, Post MJ In skeletal muscle tissue engineering, it remains a challenge to produce mature, functional muscle tissue. Mimicking the in vivo niche in in vitro culture might overcome this problem. Niche components include, for example, extracellular matrix proteins, neighbouring cells, growth factors and physical factors such as the elasticity of the matrix. Previously, we showed the effects of matrix stiffness and protein coating on proliferation and differentiation of muscle progenitor cells in a two-dimensional (2D) situation. In the present study we have investigated the additional effect of electrical stimulation. More precisely, we investigated the effect of electrical stimulation on primary myoblast maturation when cultured on top of Matrigel()- or laminin-coated substrates with varying elasticities. The effect of electrical stimulation on differentiation and maturation was found to be dependent on coating and stiffness. Although electrical stimulation enhanced myoblast maturation, the effect was mild. We therefore conclude that, with the current regimen, electrical stimulation is not essential to create functional, mature muscle tissue. Copyright (c) 2010 John Wiley & Sons, Ltd. PMID: 20603881 [PubMed - as supplied by publisher] | | |
| Tissue spheroid fusion-based in vitro screening assays for analysis of tissue maturation.
July 7, 2010 at 12:59 PM
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| | Tissue spheroid fusion-based in vitro screening assays for analysis of tissue maturation. J Tissue Eng Regen Med. 2010 Jul 5; Authors: Hajdu Z, Mironov V, Mehesz AN, Norris RA, Markwald RR, Visconti RP Organ printing or computer-aided robotic layer-by-layer additive biofabrication of thick three-dimensional (3D) living tissue constructs employing self-assembling tissue spheroids is a rapidly evolving alternative to classic solid scaffold-based approaches in tissue engineering. However, the absence of effective methods of accelerated tissue maturation immediately after bioprinting is the main technological imperative and potential impediment for further progress in the development of this emerging organ printing technology. Identification of the optimal combination of factors and conditions that accelerate tissue maturation ('maturogenic' factors) is an essential and necessary endeavour. Screening of maturogenic factors would be most efficiently accomplished using high-throughput quantitative in vitro tissue maturation assays. We have recently reviewed the formation of solid scaffold-free tissue constructs through the fusion of bioprinted tissue spheroids that have measurable material properties. We hypothesize that the fusion kinetics of these tissue spheroids will provide an efficacious in vitro assay of the level of tissue maturation. We report here the results of experimental testing of two simple quantitative tissue spheroid fusion-based in vitro high-throughput screening assays of tissue maturation: (a) a tissue spheroid envelopment assay; and (b) a tissue spheroid fusion kinetics assay. Copyright (c) 2010 John Wiley & Sons, Ltd. PMID: 20603872 [PubMed - as supplied by publisher] | | |
| Tensile strain and magnetic particle force application do not induce MAP3K8 and IL-1B differential gene expression in a similar manner to fluid shear stress in human mesenchymal stem cells.
July 7, 2010 at 12:59 PM
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| | Tensile strain and magnetic particle force application do not induce MAP3K8 and IL-1B differential gene expression in a similar manner to fluid shear stress in human mesenchymal stem cells. J Tissue Eng Regen Med. 2010 Jul 5; Authors: Glossop JR, Cartmell SH Mechanical forces, important in a variety of cellular processes, including proliferation, differentiation and gene expression, are also key in the development, remodelling and maintenance of load-bearing tissues such as cartilage and bone. Thus, there is great interest in using in vitro mechanical conditioning of mesenchymal stem cells (MSCs), multipotent adult stem cells, for tissue engineering of these tissues. In a previous gene expression study, we reported a potentially important role for mitogen-activated protein kinase kinase kinase 8 (MAP3K8) and interleukin-1beta (IL-1B) in MAPK signalling in MSCs exposed to fluid shear stress. In this follow-up study, we examined the expression of these genes in MSCs exposed to other types of mechanical force: uniaxial tensile strain (3% cell elongation) and forces generated through the exposure of magnetic particle-labelled MSCs to an oscillating magnetic field (maximum field strength 90 mT). Exposure to both types of mechanical force for 1 h did not significantly alter the gene expression of MAP3K8 or IL-1B over the 24 h period subsequent to force exposure. These data demonstrate that uniaxial tensile strain and magnetic particle-based forces do not induce MAP3K8-related MAPK signalling in the same manner as does fluid flow-induced shear stress. This illustrates divergence in the process of mechanotransduction in mechanically stimulated MSCs. Copyright (c) 2010 John Wiley & Sons, Ltd. PMID: 20603871 [PubMed - as supplied by publisher] | | |
| Using swept-source optical coherence tomography to monitor the formation of neo-epidermis in tissue-engineered skin.
July 7, 2010 at 12:59 PM
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| | Using swept-source optical coherence tomography to monitor the formation of neo-epidermis in tissue-engineered skin. J Tissue Eng Regen Med. 2010 Jul 5; Authors: Smith LE, Bonesi M, Smallwood R, Matcher SJ, Macneil S There is an increasing need for a robust, simple to use, non-invasive imaging technology to follow tissue-engineered constructs as they develop. Our aim was to evaluate the use of swept-source optical coherence tomography (SS-OCT) to image tissue-engineered skin as it developed over several weeks. Tissue-engineered skin was produced using both de-epithelialized acellular dermis (DED) and amorphous collagen gels. In both cases the epidermis could be readily distinguished from the neodermis, based on a comparison with standard destructive histology of samples. Constructs produced with DED showed more epidermal/dermal maturation than those produced using collagen. The development of tissue-engineered skin based on DED was accurately monitored with SS-OCT over 3 weeks and confirmed with conventional histology. Copyright (c) 2010 John Wiley & Sons, Ltd. PMID: 20603865 [PubMed - as supplied by publisher] | | |
| Multi-component extracellular matrices based on peptide self-assembly.
July 7, 2010 at 12:59 PM
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| | Multi-component extracellular matrices based on peptide self-assembly. Chem Soc Rev. 2010 Jul 5; Authors: Collier JH, Rudra JS, Gasiorowski JZ, Jung JP Extracellular matrices (ECMs) are challenging design targets for materials synthesis because they serve multiple biological roles, and they are composed of multiple molecular constituents. In addition, their composition and activities are dynamic and variable between tissues, and they are difficult to study mechanistically in physiological contexts. Nevertheless, the design of synthetic ECMs is a central consideration in applications such as regenerative medicine and 3D cell culture. In order to produce synthetic matrices having both multi-component construction and high levels of compositional definition, strategies based on molecular self-assembly are receiving increasing interest. These approaches are described in this tutorial review and compared with the structures and processes in native ECMs that serve as their inspiration. PMID: 20603663 [PubMed - as supplied by publisher] | | |
| Alternative splicing and muscular dystrophy.
July 7, 2010 at 12:59 PM
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| | Alternative splicing and muscular dystrophy. RNA Biol. 2010 Jul 3;7(4) Authors: Pistoni M, Ghigna C, Gabellini D Alternative splicing of pre-mRNAs is a major contributor to proteomic diversity and to the control of gene expression in higher eukaryotic cells. For this reasons, alternative splicing is tightly regulated in different tissues and developmental stages and its disruption can lead to a wide range of human disorders. The aim of this review is to focus on the relevance of alternative splicing for muscle function and muscle disease. We begin by giving a brief overview of alternative splicing, muscle-specific gene expression and muscular dystrophy. Next, to illustrate these concepts we focus on two muscular dystrophy, myotonic muscular dystrophy and facioscapulohumeral muscular dystrophy, both associated to disruption of splicing regulation in muscle. PMID: 20603608 [PubMed - as supplied by publisher] | | |
| Preparing nano-calcium phosphate particles via a biologically friendly pathway.
July 7, 2010 at 12:59 PM
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| | Preparing nano-calcium phosphate particles via a biologically friendly pathway. Biomed Mater. 2010 Jul 6;5(4):041001 Authors: Hu Q, Ji H, Liu Y, Zhang M, Xu X, Tang R It is widely agreed that nano-calcium phosphates (CaP) play an important role in tissue engineering and medical application due to their unique biological characteristics. However, the properties of nano-CaP, including bioactivity, biocompatibility and mechanical properties, are tailored over wide ranges by controlling the size and morphology of particles. Therefore, it is important to develop synthesis methods which can control the particle size distribution and shape uniformly. In this study, we report that polyacrylic acid (PAA) can act as an efficient agent to modulate nano-CaP formation. The dimension of the resultant sphere-like nanoparticles (5-60 nm) can readily be regulated by changing PAA concentrations (75-200 microg ml(-1)). In contrast to other additives, PAA is a water-soluble polymer that has already been used as an excellent biocompatible implant material in vivo. Our in vitro proliferation experiments of bone marrow mesenchymal stem cells (BMSCs) demonstrate that the involvement of PAA does not change the bioactivity of the resultant nano-CaP. In contrast, the nano-CaP fabricated by using another typical control agent, hexadecyl (cetyl) trimethyl ammonium bromide, suppressed the cell proliferation of BMSCs. Thus, we suggest that the biopolymer, PAA, can provide a more biologically friendly pathway to prepare biological nano-CaP for biomedical application. PMID: 20603529 [PubMed - as supplied by publisher] | | |
| Neuroblastoma and bone metastases: Clinical significance and prognostic value of Dickkopf 1 plasma levels.
July 7, 2010 at 12:59 PM
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| | Neuroblastoma and bone metastases: Clinical significance and prognostic value of Dickkopf 1 plasma levels. Bone. 2010 Jul 2; Authors: Granchi D, Corrias MV, Garaventa A, Baglio SR, Cangemi G, Carlini B, Paolucci P, Giunti A, Baldini N The critical role of the Wnt pathway inhibition in sustaining the onset of bone lesions has been demonstrated in a variety of bone diseases and tumors, and it has been associated with cancer aggressiveness. We have previously demonstrated that neuroblastoma cells express Dickkopf 1 (Dkk1), an inhibitor of the canonical Wnt pathway which prevents the differentiation of bone-forming cells. Since Dkk1 is a secreted factor, it could have potential clinical application as tumor marker for detecting bone metastasis and monitoring of disease. In this study, we investigated the diagnostic and prognostic value of Dkk1 plasma levels in 92 children affected by neuroblastoma, including 32 with bone metastases. Fifty-seven children hospitalized for minor surgical problems served as control group. Circulating levels of Dkk1 were higher in healthy children than in normal adults, and were comparable to those found in adult patients with aggressive tumors. No significant differences were found between neuroblastoma patients and controls, and between patients with and without bone metastases. However, when only patients with metastatic neuroblastoma were considered, the highest Dkk1 levels were detected in patients that poorly responded to induction chemotherapy and in subjects with unamplified MYCN and three or more different metastatic sites. The 'Receiver Operating Characteristic' curve enabled us to identify a threshold value to distinguish patients who were unresponsive to induction treatment. The relationship between Dkk1 and drug resistance was supported by in vitro experiments, since an increased sensitivity to doxorubicin was found in neuroblastoma cells releasing low Dkk1 levels, either constitutively or experimentally following the treatment with specific siRNA. In conclusion, Dkk1 is released by neuroblastoma cells and is able to affect the balance between osteoblastogenesis and osteoclastogenesis, thus favoring the onset of osteolytic metastases. Nevertheless, Dkk1 plasma levels do not allow the detection of bone lesions in neuroblastoma, but seem to have a predictive value with regard to the severity and the prognosis of the disease in a subset of patients with metastatic tumor. New knowledge on the biological role of Dkk1 in driving the natural history of neuroblastoma has to be further investigated, and could help to establish specific therapeutic strategies able to target key factors of tumor progression. PMID: 20603237 [PubMed - as supplied by publisher] | | |
| The influence of covalently linked and free polyethylene glycol on the structural and release properties of rhBMP-2 loaded microspheres.
July 7, 2010 at 12:59 PM
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| | The influence of covalently linked and free polyethylene glycol on the structural and release properties of rhBMP-2 loaded microspheres. J Control Release. 2010 Jul 2; Authors: Lochmann A, Nitzsche H, von Einem S, Schwarz E, Mäder K The current clinical success of therapies with recombinant human Bone Morphogenetic Protein 2 (rhBMP-2) is limited due to inefficient delivery. The high doses applied have frequently been related to severe adverse effects such as tissue swelling, seroma, inflammatory effects and heterotopic ossification. The controlled delivery of lower doses is supposed to reduce adverse effect incidence as well as costs. In this study, novel polyethylene glycol - poly(lactic-co-glycolic) acid (PEG-PLGA) diblock copolymers were used to produce low dose controlled delivery vehicles for rhBMP-2. A method to fabricate a variety of microsphere formulations with a high encapsulation efficiency in high yields was developed. The influence of PEG as an inner phase cosolvent and linked PLGA as copolymer was investigated. Six different microsphere systems with varying PEG amounts in both core and shell were characterised thoroughly with respect to the specific properties of rhBMP-2. The particle size of the microspheres was investigated with both laser diffraction and environmental scanning electron microscopy. Higher PEG/PLGA ratios showed a tendency to increase in size and a wider distribution. Due to the low rhBMP-2 doses, a profound characterisation was very challenging. The growth factor was covalently attached to rhodamine B for the first time. Studies on drug distribution in the microspheres were performed by means of confocal laser scanning microscopy. The addition of PEG to the inner phase was found to impair the formation of spherical microdomains with localized higher growth factor concentrations. Release profiles, determined with ELISA, were linked to the structural changes that were monitored. Distinct, controlled release profiles were achieved in all formulations and showed that PEG is a versatile tool in the effective control of release rates from microspheres. Higher PEG/PLGA ratios in the polymer were shown to increase the release rate from the microspheres. In contrast, PEG administered to the inner phase decreased the release rate. The biological activity of released protein was shown in vitro in an alkaline phosphatase assay. It was demonstrated that PEG-PLGA microspheres are a promising sustained delivery system which allows a reduction of the required rhBMP-2 dose to limit both adverse effects and costs. Furthermore, the data indicated that the use of PEG as an inner phase cosolvent is not suitable for rhBMP-2 in contrast the reported beneficial effects for other growth factors. PMID: 20603166 [PubMed - as supplied by publisher] | | |
| Multipotent mesenchymal stem cell grafting to treat cutaneous radiation syndrome: development of a new minipig model.
July 7, 2010 at 12:59 PM
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| | Multipotent mesenchymal stem cell grafting to treat cutaneous radiation syndrome: development of a new minipig model. Exp Hematol. 2010 Jun 25; Authors: Agay D, Scherthan H, Forcheron F, Grenier N, Hérodin F, Meineke V, Drouet M OBJECTIVES: Cutaneous radiation syndrome (CRS) is the delayed consequence of localized skin exposure to high doses of ionizing radiation (IR). Recent grafting of three IR-burned patients has suggested the benefit of local bone marrow mesenchymal stem cell (MSC) injection in favour of wound healing and pain control. Here, we have developed a new minipig model of severe CRS to study underlying mechanisms of this cell therapy approach. MATERIALS AND METHODS: Göttingen minipigs were locally irradiated using a (60)Co gamma source as follows: ungrafted 50 and 60 Gy (n=4); grafted 50 and 60 Gy (n=3). Bone marrow MSCs were cultured in MEM with 10% fetal calf serum and basic fibroblast growth factor (2ng.mL(-1)). Autologous MSCs were intradermally injected twice or three times from days 27 to 96 (range 99-128.5x10(6) MSCs per injection). RESULTS: All animals exhibited a clinical evolution similar to human after a latency phase of several weeks: early erythema, hair loss, dry/moist desquamation followed by necrosis during 81 - 222 days post IR. Skin damage in higher exposed animals appeared slightly earlier. Immuno-histology revealed severe skin damage in all animals and rhabdomyolysis in the muscle tissue below the entry area, with the latter being more severe in controls. In grafted animals, the MSCs led to local accumulation of lymphocytes at the dermis/subcutis border and improved vascularization. CONCLUSION: This study establishes a new minipig model that is close to human and allows the development of stem cell therapy strategies that may be applied in treatment of human radiation burns. PMID: 20600578 [PubMed - as supplied by publisher] | | |
| Oxygen and oxygenation in stem-cell therapy for myocardial infarction.
July 7, 2010 at 12:59 PM
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| | Oxygen and oxygenation in stem-cell therapy for myocardial infarction. Life Sci. 2010 Jun 28; Authors: Khan M, Kwiatkowski P, Rivera BK, Kuppusamy P Myocardial infarction (MI) is caused by deprivation of oxygen and nutrients to the cardiac tissue due to blockade of coronary artery. It is a major contributor to chronic heart disease, a leading cause of mortality in the modern world. Oxygen is required to meet the constant energy demands for heart contractility, and also plays an important role in the regulation of heart function. However, reoxygenation of the ischemic myocardium upon restoration of blood flow may lead to further injury. Controlled oxygen delivery during reperfusion has been advocated to prevent this consequence. Monitoring the myocardial oxygen concentration would play a vital role in understanding the pathological changes in the ischemic heart following myocardial infarction. During the last two decades, several new techniques have become available to monitor myocardial oxygen concentration in vivo. Electron paramagnetic resonance (EPR) oximetry would appear to be the most promising and reliable of these techniques. EPR utilizes crystalline probes which yield a single sharp line, the width of which is highly sensitive to oxygen tension. Decreased oxygen tension results in a sharpening of the EPR spectrum, while an increase results in widening. In our recent studies, we have used EPR oximetry as a valuable tool to monitor myocardial oxygenation for several applications like ischemia-reperfusion injury, stem-cell therapy and hyperbaric oxygen therapy. The results obtained from these studies have demonstrated the importance of tissue oxygen in the application of stem cell therapy to treat ischemic heart tissues. These results have been summarized in this review article. PMID: 20600148 [PubMed - as supplied by publisher] | | |
| Gene therapy for ischemic heart disease.
July 7, 2010 at 12:59 PM
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| | Gene therapy for ischemic heart disease. J Mol Cell Cardiol. 2010 Jul 2; Authors: Lavu M, Gundewar S, Lefer DJ Current pharmacologic therapy for ischemic heart disease suffers multiple limitations such as compliance issues and side effects of medications. Revascularization procedures often end with need for repeat procedures. Patients remain symptomatic despite maximal medical therapy. Gene therapy offers an attractive alternative to current pharmacologic therapies and may be beneficial in refractory disease. Gene therapy with isoforms of growth factors such as VEGF, FGF and HGF induces angiogenesis, decreases apoptosis and leads to protection in the ischemic heart. Stem cell therapy augmented with gene therapy used for myogenesis has proven to be beneficial in numerous animal models of myocardial ischemia. Gene therapy coding for antioxidants, eNOS, HSP, mitogen-activated protein kinase and numerous other anti apoptotic proteins have demonstrated significant cardioprotection in animal models. Clinical trials have demonstrated safety in humans apart from symptomatic and objective improvements in cardiac function. Current research efforts are aimed at refining various gene transfection techniques and regulation of gene expression in vivo in the heart and circulation to improve clinical outcomes in patients that suffer from ischemic heart disease. In this review article we will attempt to summarize the current state of both preclinical and clinical studies of gene therapy to combat myocardial ischemic disease. PMID: 20600100 [PubMed - as supplied by publisher] | | |
| Growth and Differentiation Properties of Mesenchymal Stromal Cell Populations Derived from Whole Human Umbilical Cord.
July 7, 2010 at 12:59 PM
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| | Growth and Differentiation Properties of Mesenchymal Stromal Cell Populations Derived from Whole Human Umbilical Cord. Stem Cell Rev. 2010 Jul 2; Authors: Majore I, Moretti P, Stahl F, Hass R, Kasper C Up to 2.8 x 10(7) fibroblast-like cells displaying an abundant presence of mesenchymal stem cell (MSC) markers CD73, CD90, CD105 and a low level of HLA-I expression can be isolated from one whole human umbilical cord (UC) using a simple and highly reproducible explant culture approach. Cells derived from whole UC, similar to cells collected from separate compartments of UC, display a distinct chondrogenic and adipogenic potential. Therefore they are potential candidates for cartilage and adipose tissue engineering. Cell differentiation along the osteogenic pathway is, however, less efficient, even after the addition of 1.25-dihydroxyvitamin D3, a potent osteoinductive substance. Isolated cells are highly proliferative, tolerate cryopreservation with an average survival rate of about 75% and after thawing can be propagated further, at least over 20 population doublings before their proliferative activity begins to decline. More importantly, they synthesize numerous trophic factors including neurotrophins and factors which facilitate angiogenesis and hematopoiesis. In conclusion, cells isolated from whole UC satisfies all requirements essential for the generation of stem cell banks containing permanently available cell material for applications in the field of regenerative medicine. Nevertheless, further studies are needed to improve and adjust the methods which are already employed for adult MSC expansion and differentiation to specific properties and requirements of the primitive stem cells collected from UC. So, our data verify that the choice of individual parameters for cell propagation, such as duration of cell expansion and cell seeding density, has a substantial impact on the quality of UC-derived cell populations. PMID: 20596801 [PubMed - as supplied by publisher] | | |
| Autologous matrix-induced chondrogenesis (AMIC). A one-step procedure for retropatellar articular resurfacing.
July 7, 2010 at 12:59 PM
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| | Autologous matrix-induced chondrogenesis (AMIC). A one-step procedure for retropatellar articular resurfacing. Acta Orthop Belg. 2010 Apr;76(2):260-3 Authors: Benthien JP, Behrens P The objective of this technical note is to describe the autologous matrix induced chondrogenesis (AMIC) procedure and to evaluate its possible role for resurfacing of retropatellar cartilage defects. AMIC is a one-step procedure combining microfracturing with application of a collagen I/III membrane to protect the initial blood clot and to serve as a scaffold for the developing chondrocytes. A retrospective analysis of our experience in three patients followed for 18 months is presented. PMID: 20503954 [PubMed - indexed for MEDLINE] | | |
| 5-Aza-2'-deoxycytidine treatment induces skeletal myogenic differentiation of mouse dental pulp stem cells.
July 7, 2010 at 12:59 PM
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| | 5-Aza-2'-deoxycytidine treatment induces skeletal myogenic differentiation of mouse dental pulp stem cells. Arch Oral Biol. 2010 May;55(5):350-7 Authors: Nakatsuka R, Nozaki T, Uemura Y, Matsuoka Y, Sasaki Y, Shinohara M, Ohura K, Sonoda Y OBJECTIVE: Tissue stem cells in dental pulp are assumed to possess differentiation potentials similar to mesenchymal stem cells (MSCs). The aim of this in vitro study is to examine the differentiation potentials of mouse dental pulp stem cells (DPSCs) and develop the appropriate differentiation assay systems for skeletal myogenic differentiation of these cells. METHODS: Dental pulps were extracted from mandible sections of C57/BL6 mice, and adherent dental pulp cells were isolated in culture. These cells were cultured in osteogenic or adipogenic induction medium to induce osteogenic and adipogenic differentiation. On the other hand, the skeletal myogenic differentiation potential of these cells was investigated using different conditions, such as serum-free medium, Myod1 overexpression, or 5-Aza-2'-deoxycytidine (5-Aza) treatment for DNA demethylation. Muscle-specific transcriptional factor expression was evaluated by RT-PCR, and myotube formation and myosin heavy chain expression were evaluated by phase-contrast microscopy and immunofluorescence staining, respectively. RESULTS: The adherent dental pulp cells exhibited a proliferative capacity and they showed osteogenic and adipogenic differentiation as seen in previous studies. Although the expression of Myod1 mRNA and myotube formation was not detected in serum-free conditions, the forced expression of Myod1 up-regulated the expression of Myogenin and Pax7 mRNA. However, myotube formation was not confirmed. Interestingly, myosin heavy chain expression and myotube formation were observed following 5-Aza treatment of these cells. CONCLUSIONS: These results demonstrated that mouse DPSCs possess MSC-like differentiation potential. DNA demethylation induced by 5-Aza treatment resulted in the skeletal muscle differentiation in mouse DPSCs, suggesting that DNA demethylation might trigger this differential induction of mouse DPSCs. PMID: 20362276 [PubMed - indexed for MEDLINE] | | | | 
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