J A Andrades

University of Malaga, Málaga, Andalusia, Spain

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Publications (44)126.23 Total impact

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    ABSTRACT: MicroRNAs (miRNAs), small non-coding RNAs, regulate gene expression primarily at the posttranscriptional level. We previously found that miR-335 is critically involved in the regulation and differentiation capacity of human mesenchymal stem cells (hMSCs) in vitro. In this study, we investigated the significance of miR-335 for the therapeutic potential of hMSCs. Analysis of hMSCs in ex vivo culture demonstrated a significant and progressive increase in miR-335 that is prevented by telomerase. Expression levels of miR-335 were also positively correlated with donor age of hMSCs, and were increased by stimuli that induce cell senescence, such as γ-irradiation and standard O2 concentration. Forced expression of miR-335 resulted in early senescence-like alterations in hMSCs, including: increased SA-β-gal activity and cell size, reduced cell proliferation capacity, augmented levels of p16 protein, and the development of a senescent-associated secretory phenotype (SASP). Furthermore, overexpression of miR-335 abolished the in vivo chondro-osseous potential of hMSCs, and disabled their immunomodulatory capacity in a murine experimental model of lethal endotoxemia. These effects were accompanied by a severely reduced capacity for cell migration in response to proinflammatory signals and a marked reduction in Protein Kinase D1 (PRKD1) phosphorylation, resulting in a pronounced decrease of AP-1 activity. Our results demonstrate that miR-335 plays a key role in the regulation of reparative activities of hMSCs and suggests that it might be considered a marker for the therapeutic potency of these cells in clinical applications. Stem Cells 2014
    Stem Cells 03/2014; · 7.70 Impact Factor
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    ABSTRACT: Transforming growth factor-beta (TGF-β) is involved in processes related to the differentiation and maturation of osteoprogenitor cells into osteoblasts. Rat bone marrow (BM) cells were cultured in a collagen-gel containing 0.5% fetal bovine serum (FBS) for 10 days in the presence of rhTGF (recombinant human TGF)-β1-F2, a fusion protein engineered to include a high-affinity collagen-binding decapeptide derived from von Willebrand factor. Subsequently, cells were moderately expanded in medium with 10% FBS for 4 days and treated with a short pulse of rhBMP (recombinant human bone morphogenetic protein)-2 for 4 h. During the last 2 days, dexamethasone and β-glycerophosphate were added to potentiate osteoinduction. Concomitant with an up-regulation of cell proliferation, DNA synthesis levels were determined. Polymerase chain reaction was performed to reveal the possible stemness of these cells. Osteogenic differentiation was evaluated in terms of alkaline phosphatase activity and mineralized matrix formation as well as by mRNA expression of osteogenic marker genes. Moreover, cells were placed inside diffusion chambers and implanted subcutaneously into the backs of adult rats for 4 weeks. Histological study provided evidence of cartilage and bone-like tissue formation. This experimental procedure is capable of selecting cell populations from BM that, in the presence of rhTGF-β1-F2 and rhBMP-2, achieve skeletogenic potential in vitro and in vivo.
    International Journal of Molecular Sciences 01/2014; 15(7):11255-11274. · 2.46 Impact Factor
  • Nano Research 12/2013; · 7.39 Impact Factor
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    ABSTRACT: In vivo testing is a mandatory last step in scaffold development. Agile longitudinal non-invasive real time monitoring of stem cell behaviour in biomaterials implanted in live animals should facilitate the development of scaffolds for tissue engineering. We report on a non-invasive bioluminescence imaging (BLI) procedure for simultaneous monitoring of changes in the expression of multiple genes to evaluate scaffold performance in vivo. Adipose tissue derived stromal mensenchymal cells were dually labelled with Renilla-RFP and Firefly-GFP chimeric reporters regulated by CMV and tissue specific promoters, respectively. Labelled cells were induced to differentiate in vitro and in vivo, by seeding in demineralised bone matrixes (DBMs) and monitored by BLI. Imaging results were validated by RT-PCR and histological procedures. The proposed approach improves molecular imaging and measurement of changes in gene expression of cells implanted in live animals. This procedure, applicable to the simultaneous analysis of multiple genes from cells seeded in DBMs should facilitate engineering of scaffolds for tissue repair.
    Tissue Engineering Part A 09/2012; · 4.64 Impact Factor
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    ABSTRACT: Spinal arthrodesis consists of a combination of a system of mechanical stabilisation of one or more vertebral segments with a biological substance that promotes osteoneogenesis, with aim of achieving the permanent fusion between areas more or less the same size of these segments. In spinal arthrodesis, the biological support par excellence is the autograft. However, obtaining this involves a high incidence of morbidity and, in cases of arthrodesis of more than one intervertebral space, the quantity available is usually insufficient. The extraction and implantation time prolongs the surgery, increasing the exposure to and risk of bleeding and infection. For these reasons, there is a search for substances that possess the properties of the autograft, avoiding the morbidity and added surgical time required to extract the autograft. The biomechanical-biological interaction in vertebral arthrodesis has been studied in this article.
    Revista Espanola de Cirugia Ortopedica y Traumatologia 05/2012; 56(3):227-44.
  • Revista Española de Cirugía Ortopédica y Traumatología 05/2012; 56(3):227-244.
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    ABSTRACT: Articular cartilage (AC) is an avascular tissue with precise polarity and organization. The three distinct zones are: surface, middle and deep. The production and accumulation of the superficial zone protein (SZP), also known as lubricin, by the surface zone is a characteristic feature of AC. To date, there is a wealth of evidence showing differentiation of AC from mesenchymal stem cells. Most studies that described chondrogenic differentiation did not focus on AC with characteristic surface marker SZP/lubricin. The present investigation was initiated to determine the induction of SZP/lubricin in skeletal muscle-derived mesenchymal stem/progenitor cells (MDMSCs) by transforming growth factor-β1 (TGF-β1) and bone morphogenetic protein-7 (BMP-7). MDMSCs were cultured as a monolayer at a density of 1 × 105 cells/well in 12-well tissue culture plates. Cell cultures were treated for 3, 7 and 10 days with TGF-β1 and BMP-7. The medium was analyzed for SZP. The cells were used to isolate RNA for RT-PCR assays for SZP expression. The SZP/lubricin increased in a time-dependent manner on Days 3, 7 and 10 in the medium. As early as Day 3, there was a three-fold increase in response to 3 ng/ml of TGF-β1 and 300 ng/ml of BMP-7. This was confirmed by immunochemical localization of SZP as early as Day 3 after treatment with TGF-β1. The expression of SZP mRNA was enhanced by TGF-β1. The present investigation demonstrated the efficient and reproducible induction of SZP/lubricin accumulation by TGF-β1 and BMP-7 in skeletal MDMSCs. Optimization of the experimental conditions may permit the utility of MDMSCs in generating surface zone-like cells with phenotypic markers of AC and, therefore, constitute a promising cell source for tissue engineering approaches of superficial zone cartilage.
    Arthritis research & therapy 04/2012; 14(2):R72. · 4.27 Impact Factor
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    ABSTRACT: Stem cell transplantation therapy using mesenchymal stem cells (MSCs) is considered a useful strategy. Although MSCs are commonly isolated by exploiting their plastic adherence, several studies have suggested that there are other populations of stem and/or osteoprogenitor cells which are removed from primary culture during media replacement. Therefore, we developed a three-dimensional (3D) culture system in which adherent and non-adherent stem cells are selected and expanded. Here, we described the characterization of 3D culture-derived cell populations in vitro and the capacity of these cells to differentiate into bone and/or cartilage tissue when placed inside of demineralized bone matrix (DBM) cylinders, implanted subcutaneously into the backs of rat for 2, 4 and 8 weeks. Our results demonstrates that 3D culture cells were a heterogeneous population of uncommitted cells that express pluripotent, hematopoietic, mesenchymal and endothelial specific markers in vitro and can undergo osteogenic differentiation in vivo.
    Cell Transplantation 04/2012; · 4.42 Impact Factor
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    Regenerative Medicine and Tissue Engineering - Cells and Biomaterials, 08/2011; , ISBN: 978-953-307-663-8
  • A Hari Reddi, José Becerra, José A Andrades
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    ABSTRACT: Osteoarthritis (OA) is a major clinical and scientific challenge. The degradation of articular cartilage in the joints is a common manifestation of painful arthritis. The regeneration of articular cartilage in OA is an unmet clinical need. The assembly of articular cartilage by tissue engineering toward complete regeneration is the goal of most scientists and surgeons. The key ingredients for regeneration are signals, stem cells, and scaffolds. This brief review focuses on the scaffold, with special emphasis on hydrogels and nanomaterials for the assembly of tissue-engineered cartilage, and ultimately leading to the total regeneration of articular cartilage in the joints.
    Tissue Engineering Part B Reviews 05/2011; 17(5):301-5. · 4.64 Impact Factor
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    ABSTRACT: Articular cartilage (AC) has no or very low ability of self-repair, and untreated lesions may lead to the development of osteoarthritis. One method that has been proven to result in long-term repair or isolated lesions is autologous chondrocyte transplantation. However, first generation of these cells' implantation has limitations, and introducing new effective cell sources can improve cartilage repair. AC provides a resilient and compliant articulating surface to the bones in diarthrodial joints. It protects the joint by distributing loads applied to it, so preventing potentially damaging stress concentrations on the bone. At the same time it provides a low-friction-bearing surface to enable free movement of the joint. AC may be considered as a visco- or poro-elastic fiber-composite material. Fibrils of predominantly type II collagen provide tensile reinforcing to a highly hydrated proteoglycan gel. The tissue typically comprises 70% water and it is the structuring and retention of this water by the proteoglycans and collagen that is largely responsible for the remarkable ability of the tissue to support compressive loads.
    Tissue Engineering Part B Reviews 12/2010; 16(6):617-27. · 4.64 Impact Factor
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    ABSTRACT: Recent advances in stem cell research have highlighted the role played by such cells and their environment (the stem cell niche) in tissue renewal and homeostasis. The control and regulation of stem cells and their niche are remaining challenges for cell therapy and regenerative medicine on several tissues and organs. These advances are important for both, the basic knowledge of stem cell regulation, and their practical translational applications into clinical medicine. This article is primarily concerned with the mesenchymal stem cells (MSCs) and it reviews the current aspects of their own niche. We discuss on the need for a deeper understanding of the identity of this cell type and its microenvironment in order to improve the effectiveness of any cell therapy for regenerative medicine. Ex vivo reproduction of the conditions of the natural stem cell niche, when necessary, would provide success to tissue engineering. The first challenge of regenerative medicine is to find cells able to replace and/or repair the lost function of tissues and organs by disease or aging and the trophic and immunomodulatory effects recently found for MSCs open up for new opportunities. If MSCs are pericytes, as it has been proposed, perhaps it may explain the ubiquity of these cells and their possible role in miscellaneous repairs throughout the body opening for new chances for extensive tissue repair.
    Stem cell reviews 10/2010; 7(2):248-55. · 5.08 Impact Factor
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    ABSTRACT: The utility of recombinant human bone morphogenetic protein-2 (rhBMP-2) in inducing bone formation in fractures of bone is well known. However, the influence of the mechanical environment on the actions of rhBMP-2 on fracture healing is not clear. An experimental model of fractures of the tibia in rabbits was developed and utilized to investigate the role of mechanical environment on rhBMP-2 action. A 1 mm osteotomy gap was stabilized by either a low- or high-stiffness fixator (LSF or HSF, respectively), and local treatment with rhBMP-2 in an absorbable collagen sponge (ACS) was evaluated. The results of the investigation were analysed by both histomorphometry and biomechanics. The LSF caused an increase in mineralized periosteal callus compared to HSF, the rhBMP-2 in ACS accelerated fracture healing only in the LSF group but not in the HSF group. The area of mineralized tissue in interfragmentary callus was determined by fixation stiffness and not by BMP treatment. rhBMP-2 caused higher bone resorption in the endosteal callus during the late stages of fracture healing, but these histological differences did not affect the mechanical properties. Biomechanical evaluation showed only differences at 3 weeks between LSF-rhBMP-2 and LSF-ACS. The bending and torsional properties were higher in the rhBMP-2/ACS group compared to ACS alone at 3 weeks.
    Journal of Tissue Engineering and Regenerative Medicine 03/2010; 4(7):543-52. · 4.43 Impact Factor
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    ABSTRACT: We report the spinal results of the use of bone morphogenetic protein (BMP-7) and pluripotent stem cells differentiated to the osteogenic lineage. The results of this randomized, controlled clinical trial show that BMP-7 achieved similar bone formation to the use of autograft. Equally, in another controlled prospective cohort, greater bone formation was found when BMP-7 was used with allograft compared with allograft alone. No adverse effects were found. Experimental studies of cell therapy show promising results. The current foundations of cell therapy are discussed.
    Precambrian Research - PRECAMBRIAN RES. 01/2010; 54:11-18.
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    ABSTRACT: Non-invasive bioluminescence imaging (BLI) to monitor changes in gene expression of cells implanted in live animals should facilitate the development of biomaterial scaffolds for tissue regeneration. We show that, in vitro, induction of chondrogenic differentiation in mouse bone marrow stromal cell line (CL1) and human adipose tissue derived mesenchymal stromal cells (hAMSCs), permanently transduced with a procollagen II (COL2A1) promoter driving a firefly luciferase gene reporter (PLuc) (COL2A1p.PLuc), induces PLuc expression in correlation with increases in COL2A1 and Sox9 mRNA expression and acquisition of chondrocytic phenotype. To be able to simultaneously monitor in vivo cell differentiation and proliferation, COL2A1p.PLuc labelled cells were also genetically labelled with a renilla luciferase (RLuc) gene driven by a constitutively active cytomegalovirus promoter, and then seeded in demineralized bone matrix (DBM) subcutaneously implanted in SCID mice. Non-invasive BLI monitoring of the implanted mice showed that the PLuc/RLuc ratio reports on gene expression changes indicative of cell differentiation. Large (CL1) and moderated (hAMSCs) changes in the PLuc/RLuc ratio over a 6 week period, revealed different patterns of in vivo chondrogenic differentiation for the CL1 cell line and primary MSCs, in agreement with in vitro published data and our results from histological analysis of DBM sections. This double bioluminescence labelling strategy together with BLI imaging to analyze behaviour of cells implanted in live animals should facilitate the development of progenitor cell/scaffold combinations for tissue repair.
    Biomaterials 07/2009; 30(28):4986-95. · 8.31 Impact Factor
  • S Claros, M Alonso, J Becerra, J A Andrades
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    ABSTRACT: Bone marrow (BM) has been long established as the main source of pluripotential mesenchymal stem cells (MSCs), and has been so far the main recognized source of osteoprogenitor cells that contribute to the turnover of the skeletal scaffold. The existence of an osteoprogenitor cell in other connective tissues such as skeletal muscle has been reported. In light of its availability and because of the relative ease of muscle cell isolation, skeletal muscle is an attractive source of cells for use in tissue engineering applications. The aim of this study was to explore the potential to differentiate into the chondro--osteoblastic lineage of a plastic adhering cell population, referred t as skeletal muscle-derived cells (SMDCs), obtained from biopsies of rat skeletal muscle. SMDCs displayed a fibroblast-like morphology. Our study revealed that the isolated cell population had a mesenchymal origin as indicated by abundant expression of STRO-1 and CD166. Osteogenic markers like osteocalcin (OC), bone sialoprotein (BSP) and osteopontin (OP) gene expressions were detected by RT-PCR. When these cells were cultured in the presence of an osteo-inductive culture medium, positive staining for alkaline phosphatase (ALP) and formation of mineralized matrix were increased. Furthermore SMDCs formed bone and cartilage tissues in vivo when placed inside of diffusion chambers and in demineralized bone matrix (DBM) cylinders, implanted subdermically into the backs of rat for 28 days. In conclusion, this experimental procedure is capable of selecting a cell population obtained from the skeletal muscle that is able to complete the differentiation pathway leading to the formation of cartilage and bone. In this respect SMDCs resemble BM stromal cells (BMSCs) and have demonstrated a potential application for cartilage and bone tissue engineering.
    Cellular and molecular biology (Noisy-le-Grand, France) 02/2008; 54(1):1-10. · 1.46 Impact Factor
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    ABSTRACT: Regeneration takes place in the body at a moment or another throughout life. Bone, cartilage, and tendons (the key components of the structure and articulation in the body) have a limited capacity for self-repair and, after traumatic injury or disease, the regenerative power of adult tissue is often insufficient. When organs or tissues are irreparably damaged, they may be replaced by an artificial device or by a donor organ. However, the number of available donor organs is considerably limited. Generation of tissue-engineered replacement organs by extracting stem cells from the patient, growing them and modifying them in clinical conditions after re-introduction in the body represents an ideal source for corrective treatment. Mesenchymal stem cells (MSCs) are the multipotential progenitors that give rise to skeletal cells, vascular smooth muscle cells, muscle (skeletal and cardiac muscle), adipocytes (fat tissue) and hematopoietic (blood)-supportive stromal cells. MSCs are found in multiple connective tissues, in adult bone marrow, skeletal muscles and fat pads. The wide representation in adult tissues may be related to the existence of a circulating blood pool or that MSCs are associated to the vascular system.
    Cellular and molecular biology (Noisy-le-Grand, France) 02/2008; 54(1):40-51. · 1.46 Impact Factor
  • M Alonso, S Claros, J Becerra, J A Andrades
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    ABSTRACT: Recent studies have demonstrated that adipose-derived adult stromal cells (ADASCs) offer great promise for cell-based therapies due to their ability to differentiate towards bone, cartilage and fat [corrected] The objective of this study was to investigate whether type I collagen would elicit in vivo bone formation of passaged rat adipose-derived adult stromal cells (ADASC) placed extraskeletally. After expansion for 1-4 passages (P), cells were incubated in osteogenic medium containing dexamethasone, ascorbic acid and beta-glycerol phosphate for 2-4 weeks. Undifferentiated cells were maintained in Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum (FBS). Osteogenic differentiation was evaluated by alkaline phosphatase (ALP) and von Kossa staining as well as by gene expression of ALP, osteopontin (OP), osteonectin (ON), osteocalcin (OC), collagen I (colI), collagen II (colII), bone sialoprotein (BSP), periostin (Postn), runx2, osterix (Osx), sox9, msx1 and msx2. Diffusion chambers were filled with 1x10(6) cells mixed with or without type I collagen gel and implanted subcutaneously into rats. Controls included chambers exposed to (1) undifferentiated cells (with or without collagen, (2) collagen without cells and (3) empty chambers (n=5 per group). Four weeks after implantation, in vivo bone and cartilage formation was demonstrated in implants containing 4-week osteo-induced P1 and P4 cells wrapped in the collagen gel, as confirmed by Goldner's trichrome and Alcian blue staining, respectively. Newly formed bone stained positive for type I collagen. Control implants had no bone or cartilage and were primarily filled with fibrous tissue at that time interval. Recent studies have demonstrated that ADASC offer great promise for cell-based therapies because of their ability to differentiate toward bone, cartilage and fat. However, the influence of different matrices on the in vivo osteogenic capability of ADASC is not fully understood. These findings suggest that type I collagen may support the survival and expression of osteogenic and chondrogenic phenotypes in passaged rat ADASC in vivo.
    Cytotherapy 02/2008; 10(6):597-610. · 3.06 Impact Factor
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    ABSTRACT: Recent studies on the morphogenesis of the fins of Danio rerio (zebrafish) during development and regeneration suggest that a number of inductive signals involved in the process are similar to some of those that affect bone and cartilage differentiation in mammals and humans. Akimenko et al. (2002) has shown that bone morphogenetic protein-2b (BMP2b) is involved in the induction of dermal bone differentiation during fin regeneration. Many other groups have also shown that molecules from the transforming growth factor-beta superfamily (TGFb), including BMP2, are effective in promoting chondrogenesis and osteogenesis in vivo in higher vertebrates, including humans. In the present study, we review the state of the art of this topic by a comparative analysis of skeletal tissue development, regeneration and renewal processes in tetrapods, and fin regeneration in fishes. A general conclusion of this study states that lepidotrichia is a special skeletal tissue different to cartilage, bone, enamel, or dentine in fishes, according to its extracellular matrix (ECM) composition. However, the empirical analysis of inducing signals of skeletal tissues in fishes and tetrapods suggests that lepidotrichia is different to any responding features with main skeletal tissues. A number of new inductive molecules are arising from fin development and regeneration studies that might establish an empirical basis for further molecular approaches to mammal skeletal tissues differentiation. Despite the tissue dissimilarity, this empirical evidence might finally lead to clinical applications to skeletal disorders in humans.
    The Scientific World Journal 02/2007; 7:1114-27. · 1.73 Impact Factor
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    ABSTRACT: The computational power and memory bandwidth of graphics processing units (GPUs) have turned them into attractive platforms for general-purpose applications. In this paper, we exploit this power in the context of biomedical image processing by establishing a cooperative environment between the CPU and the GPU. We deal with phenotype and color analysis on a wide variety of microscopic images from studies of cartilage and bone tissue regeneration using stem cells and genetics involving cancer pathology. Both processors are used in parallel to map algorithms for computing color histograms, contour detection using the Canny filter and pattern recognition based on the Hough transform. Task, data and instruction parallelism are exploited in the GPU to accomplish performance gains between 4times and 100times more than the typical CPU code.
    Proceedings of the 7th IEEE International Conference on Bioinformatics and Bioengineering, BIBE 2007, October 14-17, 2007, Harvard Medical School, Boston, MA, USA; 01/2007

Publication Stats

472 Citations
126.23 Total Impact Points

Institutions

  • 1994–2014
    • University of Malaga
      • • Faculty of Science
      • • Departamento de Biología Celular, Genética y Fisiología
      Málaga, Andalusia, Spain
  • 2012
    • Hospital Costa del Sol
      Marbella, Andalusia, Spain
  • 2011
    • University of California, Davis
      • Department of Orthopaedic Surgery
      Davis, CA, United States
  • 2010
    • Centro de Investigación Biomédica en Red en Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN)
      Caesaraugusta, Aragon, Spain
  • 1999
    • University of Southern California
      • Department of Medicine
      Los Angeles, CA, United States
  • 1998
    • Austral University of Chile
      • Instituto de Anatomía, Histología y Patología
      Puerto Montt, Region de Los Lagos, Chile