Kizhner T.,University of Sfax |
Ben-David D.,University of Sfax |
Rom E.,ProCore LTD |
Yayon A.,ProCore LTD |
Livne E.,University of Sfax
In Vitro Cellular and Developmental Biology - Animal | Year: 2011
Bone repair is a major concern in reconstructive surgery. Transplants containing osteogenically committed mesenchymal stem cells (MSCs) provide an alternative source to the currently used autologous bone transplants which have limited supply and require additional surgery to the patient. A major drawback, however is the lack of a critical mass of cells needed for successful transplantation. The purpose of the present study was to test the effects of FGF2 and FGF9 on expansion and differentiation of MSCs in order to establish an optimal culture protocol resulting in sufficient committed osteogenic cells required for successful in vivo transplantation. Bone marrow-derived MSCs cultured in αMEM medium supplemented with osteogenic supplements for up to three passages (control medium), were additionally treated with FGF2 and FGF9 in various combinations. Cultures were evaluated for viability, calcium deposition and in vivo osteogenic capacity by testing subcutaneous transplants in nude mice. FGF2 had a positive effect on the proliferative capacity of cultured MSCs compared to FGF9 and control medium treated cultures. Cultures treated with FGF2 followed by FGF9 showed an increased amount of extracted Alizarin red indicating greater osteogenic differentiation. Moreover, the osteogenic capacity of cultured cells transplanted in immunodeficient mice revealed that cells that were subjected to treatment with FGF2 in the first two passages and subsequently to FGF9 in the last passage only, were more successful in forming new bone. It is concluded that the protocol using FGF2 prior to FGF9 is beneficial to cell expansion and commitment, resulting in higher in vivo bone formation for successful bone tissue engineering. © 2011 The Society for In Vitro Biology.
Agency: European Commission | Branch: FP7 | Program: CP-IP | Phase: HEALTH-2009-1.4-1;HEALTH-2009-1.4-3 | Award Amount: 12.08M | Year: 2010
Osteoarthritis (OA) is a degenerative joint disease for which no efficient therapy is available. The ADIPOA consortium has previously shown that intraarticular injections of stromal cells prevents OA in two different models, but the mechanisms of this chondroprotective effect remains unknown, and the activation of cartilage derived endogenous stem cells is suspected. The participants have experience on cell therapy, logistic and production facilities for adipose derived stromal cells (ASC) and clinical studies focusing on cartilage repair. They have shown that stromal cells have anti-inflammatory and antiapoptosis effets, prevent cells from senescence and protect endogenous cells from oxidative stress. ASC are well described, and the procedure to expand the ASC in GMP clinical grade by one of us has been approved by regulatory authorities. This has prompted us to propose an original and innovative regenerative medicine approach for OA in a four years programme organised around 6 workpackages : WP 1 ASC biology, cell processing & optimization for chondral protection WP 2 In vivo validation of chondroprotective effect of ASC WP 3 Safety, Security & regulatory issues WP 4 Clinical trial endogenous ASC injected intraarticular in OA WP 5 Management and Coordination WP 6 Training and Education ADIPOA project will then lead to the optimisation and standardisation of production procedures. In vivo validation and optimisation shall lead to a phase 1 clinical trial and the design and initiation of a phase 2 controlled study in OA. The ADIPOA Consortium comprises 10 academic and 2 sme participants and gathers researchers and clinicians with expertise in clinical research, chondrocytes and adipose stem cell biology and regenerative medicine. The Sme participation will ensure the dissemination of the project results. The critical mass achieved by the ADIPOA Consortium should enable clinical applications in OA.
Agency: European Commission | Branch: FP7 | Program: CP-FP | Phase: NMP-2009-2.3-1 | Award Amount: 5.18M | Year: 2011
The intervertebral discs form the elastic part of the spine. It is composed of the annulus fibrosus, a tough outer layer of fibrocartilage, surrounding an elastic gelatinous core, the nucleus pulposus (NP). With age, the water content of the NP decreases, thus, the mechanical loads concentrate on the annulus. This leads to the NP wear, and cracking with a subsequent inflammation reaction and a prolapsed intervertebral disc. The process forms a cycle of accelerated DDD pathology. The Gold Standard for treatment is the spinal fusion, an extensive surgery, which blocks definitively free spine motion. Surgeons seek new technologies to allow motion preservation, with long-term outcomes. Based on electrospinning proprietary technology of partner NIC and a novel chemically modified ECM-based biopolymer, developed by partner ProCore, the NPmimetic consortium will develop biomimetic nano-polymer based gel for minimally invasive treatment for disc regeneration: Electrospinning technology will be exploited to design and develop nano-fiber based, biocompatible, biodegradable, synthetic scaffold that will mimic mechanical properties of native NP for immediate and short term treatment. Anti-inflammatory drugs will be carried by biodegradable nano-fibers to be gradually released in situ thus, healing and preventing inflammation. Furthermore, the synthetic scaffold will be integrated with the bioactive-polymer that is highly potent in supporting NP cells for long-term cure. A multidisciplinary study will answer scientific and engineering questions raised by the NPmimetic approach, e.g. hydrogel swelling characteristics, drug delivery, and NP cells reaction to the biomimetic gel environment. All will be supervised by a strong leader in spine surgery to define inputs and outputs of the research, from a clinical implementation point of view.
Bono F.,Sanofi S.A. |
De Smet F.,Catholic University of Leuven |
De Smet F.,Vesalius Research Center |
Herbert C.,Sanofi S.A. |
And 66 more authors.
Cancer Cell | Year: 2013
Receptor tyrosine kinases (RTK) are targets for anticancer drug development. To date, only RTK inhibitors that block orthosteric binding of ligands and substrates have been developed. Here, we report the pharmacologic characterization of the chemical SSR128129E (SSR), which inhibits fibroblast growth factor receptor (FGFR) signaling by binding to the extracellular FGFR domain without affecting orthosteric FGF binding. SSR exhibits allosteric properties, including probe dependence, signaling bias, and ceiling effects. Inhibition by SSR is highly conserved throughout the animal kingdom. Oral delivery of SSR inhibits arthritis and tumors that are relatively refractory to anti-vascular endothelial growth factor receptor-2 antibodies. Thus, orally-active extracellularly acting small-molecule modulators of RTKs with allosteric properties can be developed and may offer opportunities to improve anticancer treatment. © 2013 Elsevier Inc.
Agency: European Commission | Branch: H2020 | Program: MSCA-ITN-ETN | Phase: MSCA-ITN-2014-ETN | Award Amount: 3.97M | Year: 2015
Mobility, important for well-being, is seriously impaired by chronic low back pain and osteoarthritis in many people due to degeneration of cartilaginous tissue of the intervertebral disc and joint. To develop a treatment for these diseases this ETN aims to combine expertise in novel highly advanced drug delivery carriers with dedicated targeting tools, state of the art imaging techniques and expertise in stem cell and joint biology by training 15 young scientists in 12 partner institutes located in 5 different countries. We aim to achieve regeneration of damaged and degenerated tissues by employing targeting strategies tailored both to the pathology and the tissues involved. Regeneration of diseased tissues will be achieved by loading biologically active agents in state-of-the-art nanocarriers. The biologically active agents will stimulate the bodys own capacity to regenerate by attracting local stem cells or inhibit degeneration. Targeting will be achieved by A] injection with synthetic or natural hydrogels loaded with the nanocarriers or B] coupling diseased tissue-specific antibodies and specific hyaluronic acid moieties to the nanocarriers. Delivery and retention will be monitored by advanced in vivo and molecular imaging techniques to monitor distribution of the delivered compounds at the tissue level, as well as detect biological markers of regeneration. Major objectives: 1] To establish a network of scientists skilled in the use of smart nanocarriers, unique approach of targeting by disease-specific molecules and application of innovative imaging tools. Supported by generic scientific and training in economical and clinical valorisation, these researchers can further implement these technologies in the musculoskeletal or other areas, both in academia and industry. 2] To develop strategies exclusively targeting diseased tissues with controlled doses of bio-actives, circumventing the disadvantages of the current shotgun approaches in regenerative medicine.
Correa D.,Case Western Reserve University |
Somoza R.A.,Case Western Reserve University |
Lin P.,Case Western Reserve University |
Greenberg S.,Case Western Reserve University |
And 5 more authors.
Osteoarthritis and Cartilage | Year: 2015
Objective: To test the effects of sequential exposure to FGF2, 9 and 18 on human Mesenchymal Stem Cells (hMSC) differentiation during invitro chondrogenesis. Design: Control and FGF2-expanded hMSC were cultured in aggregates in the presence of rhFGF9, rhFGF18 or rhFGFR3-specific signaling FGF variants, starting at different times during the chondroinductive program. Quantitative real time polymerase chain reaction (qRT-PCR) and immunocytochemistry were performed at different stages. The aggregate cultures were switched to a hypertrophy-inducing medium along with rhFGFs and neutralizing antibodies against FGFR1 and FGFR3. Histological/immunohistochemical/biochemical analyses were performed. Results: FGF2-exposed hMSC during expansion up-regulated Sox9 suggesting an early activation of the chondrogenic machinery. FGF2, FGF9 and 18 modulated the expression profile of FGFR1 and FGFR3 in hMSC during expansion and chondrogenesis. In combination with transforming growth factor-beta (TGF-β), FGF9 and FGF18 inhibited chondrogenesis when added at the beginning of the program (≤d7), while exhibiting an anabolic effect when added later (≥d14), an effect mediated by FGFR3. Finally, FGFR3 signaling induced by either FGF9 or FGF18 delayed the appearance of spontaneous and induced hypertrophy-related changes. Conclusions: The stage of hMSC-dependent chondrogenesis at which the growth factors are added impacts the progression of the differentiation program: increased cell proliferation and priming (FGF2); stimulated early chondrogenic differentiation (TGF-β, FGF9/FGF18) by shifting the chondrogenic program earlier; augmented extracellular matrix (ECM) production (FGF9/FGF18); and delayed terminal hypertrophy (FGF9/FGF18). Collectively, these factors could be used to optimize pre-implantation conditions of hMSC when used to engineer cartilage grafts. © 2014 Osteoarthritis Research Society International.
PubMed | Case Western Reserve University and ProCore Ltd.
Type: Journal Article | Journal: Osteoarthritis and cartilage | Year: 2015
To test the effects of sequential exposure to FGF2, 9 and 18 on human Mesenchymal Stem Cells (hMSC) differentiation during in vitro chondrogenesis.Control and FGF2-expanded hMSC were cultured in aggregates in the presence of rhFGF9, rhFGF18 or rhFGFR3-specific signaling FGF variants, starting at different times during the chondroinductive program. Quantitative real time polymerase chain reaction (qRT-PCR) and immunocytochemistry were performed at different stages. The aggregate cultures were switched to a hypertrophy-inducing medium along with rhFGFs and neutralizing antibodies against FGFR1 and FGFR3. Histological/immunohistochemical/biochemical analyses were performed.FGF2-exposed hMSC during expansion up-regulated Sox9 suggesting an early activation of the chondrogenic machinery. FGF2, FGF9 and 18 modulated the expression profile of FGFR1 and FGFR3 in hMSC during expansion and chondrogenesis. In combination with transforming growth factor-beta (TGF-), FGF9 and FGF18 inhibited chondrogenesis when added at the beginning of the program ( d7), while exhibiting an anabolic effect when added later (d14), an effect mediated by FGFR3. Finally, FGFR3 signaling induced by either FGF9 or FGF18 delayed the appearance of spontaneous and induced hypertrophy-related changes.The stage of hMSC-dependent chondrogenesis at which the growth factors are added impacts the progression of the differentiation program: increased cell proliferation and priming (FGF2); stimulated early chondrogenic differentiation (TGF-, FGF9/FGF18) by shifting the chondrogenic program earlier; augmented extracellular matrix (ECM) production (FGF9/FGF18); and delayed terminal hypertrophy (FGF9/FGF18). Collectively, these factors could be used to optimize pre-implantation conditions of hMSC when used to engineer cartilage grafts.