Bio-Medical Materials and Engineering - Volume 18, issue s1

Authors: Stoltz, J.-F. | Netter, P. | Schmutz, J.L. | Muller, S.

Article Type: Other

Citation: Bio-Medical Materials and Engineering, vol. 18, no. s1, pp. 1-1, 2008

Authors: Sensebé, Luc

Article Type: Research Article

Abstract: Mesenchymal Stem Cells (MSCs) are multipotent adult stem cells having an immunosuppressive effect. These characteristics lead to an increasing use of MSC in graft process or for regenerative medicine. For the clinical uses of MSCs, standards are needed. The clinical grade production necessitates adhering to good manufacturing practices (GMP) to insure the delivery of a “cell drug” that is safe, reproducible and efficient. All parts of the process must be defined: the starting material (tissue origin, separation or enrichment procedures), cell density in culture, and medium (fetal calf serum (FCS) or human serum, cytokines with serum-free medium for target). …But to reach the GMP goal, cells have to be cultured in as close to a closed system as possible. Analytical methods are needed to assay the active compound and impurities. At a minimum, quality control (QC) of cells must consider the phenotype, functional potential, microbiological safety, and ensure the cultured cells remain untransformed. Finally, quality assurance system (QA) procedures specific to the production of MSCs as a cell drug must be determined and implemented. Show more

Citation: Bio-Medical Materials and Engineering, vol. 18, no. s1, pp. 3-10, 2008

Authors: De Bari, Cosimo | Dell'Accio, Francesco

Article Type: Research Article

Abstract: Cell therapies are unique in that the active component consists of living cells, which are difficult to define in their pharmacologic characteristics, and which produce variable and largely unknown amounts of bioactive molecules. Thus, the definition of the composition of a cellular product, mechanisms of action, pharmacokinetics, toxicity and efficacy assessment represent challenges never previously faced by traditional pharmacology. A pressing need for a routine use of cell therapies in the clinic is the development of quality controls for efficacy on the basis of clinically relevant potency assays, with prospective validation in human clinical trials. This review will focus on …cell-based protocols for joint surface repair. In particular, we will present the case of autologous chondrocyte implantation as an example of advanced tissue engineering technology in the clinic, with the assumption that many issues discussed can be extrapolated to other cell-based approaches in regenerative medicine. Show more

Keywords: Regenerative medicine, stem cells, cartilage repair, tissue engineering

Citation: Bio-Medical Materials and Engineering, vol. 18, no. s1, pp. 11-17, 2008

Authors: Decot, V. | Latger-Cannard, V. | Lecompte, T. | Clément, L. | Salmon, A. | Bordigoni, P. | Stoltz, J.-F. | Bensoussan, D.

Article Type: Research Article

Abstract: Chimerism analysis has become an important tool to manage patients in the peri-transplant period of allogenic stem cell transplantation. During this period, cells of donor and host origin can coexist and increasing proportion of cells of host origin is considered as a recurrence of the underlying disease. We currently performed chimerism analysis on separate peripheral blood cell subsets, lymphocytes and granulocytes. To improve our isolation method, a new automated device from Stem Cell Technology Robosep™ was tested and compared to our manual separation technique. The results obtained on T cell purification showed an improvement of the purity (98.42% with …Robosep vs. 92.42% with the manual technique Rosettesep) and of the recovery (63.43% with Robosep and 38% with Rosettesep). The results were significantly improved on patient samples with less than 10% CD3 positive cells (purity: 90% vs. 44.44%; recovery: 73.79% vs. 43.98%). Granulocytes separation was based on CD15 expression. The results showed an improvement of the purity with Robosep (96.90% vs. 86.20% with the manual technique Polymorphprep) but the recovery was impaired (35.2% vs. 52.30%). Using a myeloid (CD66/CD33) cocktail, recovery was improved with the Robosep device (64.04% with the myeloid cocktail vs. 22.4% with the CD15 cocktail). Our data demonstrated that Robosep allowed a performant cell purification in the early period post-transplantation even for populations representing less than 10% of the peripheral blood cells. Show more

Keywords: Chimerism, hematopoietic stem cell transplantation, cell subset isolation

Citation: Bio-Medical Materials and Engineering, vol. 18, no. s1, pp. 19-26, 2008

Authors: Rouy, Didier | Lebrun, Frederic | Berchem, Guy | Delagardelle, Charles | Beissel, Jean | Wagner, Daniel R.

Article Type: Research Article

Abstract: Intracoronary infusion of autologous bone marrow cells (CTX) has been shown to improve myocardial function in post infarct patients and in patients with chronic ischemic cardiomyopathy. Whether CTX affects exercise-induced changes in cardiac deformation and mitral regurgitation (MR) in patients with end stage heart failure has not been studied. In this small pilot study, eleven patients with chronic ischemic cardiomyopathy, ejection fraction (EF) <25%, no inducible ischemia and heart failure class NYHA III underwent CTX. Symptom-limited bicycle exercise echocardiography was performed pre- and 4 months post CTX and maximum systolic strain (msyε), peak systolic strain rate (psysr) and effective …regurgitant orifice of MR (ERO) were determined. There were no complications related to the procedure. The overall clinical benefit of CTX was limited with a trend towards improvement (NYHA 3.0±0.1 pre and 2.7±0.2 post CTX, p=0.06). The EF did not improve after CTX. The wall motion score index (WMSI) did not change at rest but decreased significantly during exercise (1.48±0.16 vs. 1.44±0.17, p=0.01). In conclusion, CTX may improve cardiac deformation and MR during exercise in patients with severe chronic heart failure when viable areas are targeted. Show more

Keywords: Cell therapy, heart failure, stress echocardiography, bone marrow cells

Citation: Bio-Medical Materials and Engineering, vol. 18, no. s1, pp. 27-31, 2008

Authors: Chajra, H. | Rousseau, C.F. | Cortial, D. | Ronzière, M.C. | Herbage, D. | Mallein-Gerin, F. | Freyria, A.M.

Article Type: Research Article

Abstract: Articular cartilage has a limited capacity for self-repair after trauma. Besides the conventional surgical techniques for repairing such defects, treatments involve implantation of autologous cells in suspension or within a variety of cell carrying scaffolds such as hyaluronic acid, alginate, agarose/alginate, fibrin or collagen. For the repair of full-thickness osteochondral defects, tissue engineers started to design single- or bi-phased scaffold constructs often containing hydroxyapatite-collagen composites, usually used as a bone substitute. The purpose of this study was to compare the behavior of bovine chondrocytes cultured in collagen-based scaffolds containing or not hydroxyapatite and cross-linked following two different methods. Calf chondrocytes …seeded within Hemotèse® and Collapat® II sponges (SYMATESE biomaterials), chemically cross-linked with glutaraldehyde or EDC/NHS, were maintained up to one month in culture. The cells exhibited a similar behavior in the four scaffolds regarding proliferation level, deposition of glycosaminoglycans in the scaffolds and gene expression of types I, II and X collagens, aggrecan, MMP-1, -13 and the integrin subunits α10 and α11. Show more

Citation: Bio-Medical Materials and Engineering, vol. 18, no. s1, pp. 33-45, 2008

Authors: Wang, Yun | de Isla, Natalia | Huselstein, Céline | Wang, Binghua | Netter, Patrick | Stoltz, Jean-François | Muller, Sylvaine

Article Type: Research Article

Abstract: To investigate whether the application of alginate culture and mechanical stimulation will improve the synthesis of cartilaginous matrix in dedifferentiated chondrocytes, rat chondrocytes underwent dedifferentiation upon serial monolayer culture up to passage 6, and then were encapsulated in 2% alginate gel and subject to static culture. After 28 days culture in static, the beads were exposed to 48 h of mechanical stimulation with continuous agitation. The sGAG content in alginate bead was measured by alcian blue staining. The expression of collagen protein was detected using immunofluorescence. After 28 days culture in alginate bead, the dedifferentiated chondrocytes remained round in shape …and re-synthesized the chondrocyte-specific matrix. Compared with static culture, mechanical stimulation induced statistically increases in the production of glycosaminoglycan (p≤0.01), as well as in the synthesis of collagen type II protein (p≤0.05). On the contrary, no positive expression of collagen type I protein was observed at the end of culture. Our results demonstrated that both of alginate culture and mechanical stimulation help to restore chondrocyte phenotype and promotes the synthesis of cartilaginous matrix. Show more

Keywords: Alginate, mechanical stimulation, matrix, chondrocyte, cartilage tissue engineering

Citation: Bio-Medical Materials and Engineering, vol. 18, no. s1, pp. 47-54, 2008

Authors: Schulz, R.M. | Zscharnack, M. | Hanisch, I. | Geiling, M. | Hepp, P. | Bader, A.

Article Type: Research Article

Abstract: In this study we compared the in vitro formation of cartilaginous grafts composed of collagen type I hydrogel with both ovine primary articular chondrocytes (AC) and bone marrow derived mesenchymal stem cells (MSC). During 4 weeks of culture, aggregate properties were quantitatively verified, cell viability and the expression of cartilage markers were assayed. Different microscopic techniques indicated a subdivision of MSC based scaffolds into a central construct region with uniformly distributed stem cells with low levels of apoptosis, and peripheral layers of proliferative cells, which undergo differentiation. Immunohistochemical staining and quantitative measurements of sulfated glycosaminoglycans (s-GAG) of MSC hydrogels showed …a significant increase in matrix deposition, mainly in outer areas. Furthermore, semi-quantitative RT-PCR of MSC specimens reflected a constant collagen type I activity with enhanced collagen type II, aggrecan and Sox9 expression which would suggest hyaline-like cartilage formation. We propose the application of MSC seeded grafts for the repair of focal cartilage lesions. Show more

Citation: Bio-Medical Materials and Engineering, vol. 18, no. s1, pp. 55-70, 2008

Authors: Bieback, Karen | Kern, Susanne | Kocaömer, Asli | Ferlik, Kathrin | Bugert, Peter

Article Type: Research Article

Abstract: Mesenchymal stromal cells (MSC) are currently in focus because of their clinical potential in cell therapy and tissue engineering. As yet bone marrow represents the main source of MSC for both experimental and clinical studies. However, it is speculated that the clinical value might be diminished as both the number of MSC and their differentiation capacity decline with age. Alternatively, MSC have been successfully isolated from nearly every tissue attempted so far. Our work is focused on comparing MSC derived from human adult bone marrow, lipoaspirate as well as cord blood in terms of being alternatives containing high precursor frequencies …and youngest adult cells. Applying identical culture conditions, major differences were observable in the frequencies and expansion potential, whereas basic biological features were comparable. Since all three tissues have been shown to contain multipotential cells we consider aspects such as isolation efficacy, frequency and expansion potential may more likely affect MSC clinical exploitation. Show more

Citation: Bio-Medical Materials and Engineering, vol. 18, no. s1, pp. 71-76, 2008

Authors: Kadi, A. | Fawzi-Grancher, S. | Lakisic, G. | Stoltz, J.F. | Muller, S.

Article Type: Research Article

Abstract: Tissue engineering requires the response of the cells to different stimuli inducing the synthesis of the extracellular matrix (ECM). It was been shown that mechanical and biochemical stimuli acted on the synthesis of ECM, particularly type I and III collagens. Growth factors implied in transduction pathways are multiple, but the main is TGF-β. Member of the transforming growth factor-β (TGF-β) family bind to type II and type I serine/threonine kinase receptors, which initiate intracellular signals through activation of SMADs proteins. Nevertheless, the effects of mechanical stress of this pathway remain unknown. The aim of this work was to study …the pathway of TGF-β via the SMADs proteins under mechanical (stretching) and biochemical (TGF-β) stimulations. Endogenous SMADs expression and its modulation by biochemical and mechanical stimulations were evaluated by both flow cytometry and confocal microscopy. Our results demonstrate that 10 ng of TGF-β and stretching (5%, 1 Hz) applied during 15 min induced a negative feed back loop which blocks the signalling pathway to control TGF-β activity. This inhibition effect was raised after 1 h of stimulation. Nevertheless, these preliminary studies should be continued by study of expression and localization of inhibitory SMADs (SMAD7). Show more

Keywords: Signal transduction, TGF-β, SMAD proteins, stretching stimulation, fibroblasts

Citation: Bio-Medical Materials and Engineering, vol. 18, no. s1, pp. 77-86, 2008

Authors: Builles, Nicolas | Bechetoille, Nicolas | Justin, Virginie | André, Valérie | Burillon, Carole | Damour, Odile

Article Type: Research Article

Abstract: To reconstruct artificial stroma close to corneal stroma, it is necessary to use keratocytes with high proliferative potential that maintain the keratocyte phenotype as characterised by CD34. To select such cells, we tested the proliferative potential and characterised the keratocytes isolated from 4 different areas of the human cornea: superior perilimbal, inferior perilimbal, superior central and inferior central. Keratocytes isolated from these different areas had significantly different growth rates (p<0.05), as measured by population doublings: superior perilimbal (42.59±11.78) > inferior perilimbal (38.23±12.67) > superior central (35.69±8.07) > inferior central (25.35±7.63). Their clonogenic potential evolved in the same order. Moreover, CD34 …labelling gave higher levels in the central areas in relation to the perilimbal areas. We found the best location for isolating keratocytes for stromal reconstruction. The superior perilimbal area had the greatest capacity for proliferation, as well as the best clonogenic potential and the average CD34 level (70%) remained high. Show more

Keywords: Keratocytes, culture, localisation

Citation: Bio-Medical Materials and Engineering, vol. 18, no. s1, pp. 87-98, 2008

Authors: de Isla, N.G. | Mainard, D. | Muller, S. | Stoltz, J.F.

Article Type: Research Article

Abstract: Osteoarthritis (OA) is a progressive joint disease which represents a combination of several disorders leading to cartilage degradation. The production of nitric oxide (NO) by iNOS, which is stimulated by proinflammatory mediators, is involved in cartilage catabolism and should therefore be considered a potential target for therapeutic interventions. Diacerein is a slow-acting drug that may slow down the breakdown of cartilage and relieve pain and swelling. Diacerhein, in contrast to an NSAID, is a potent inhibitor of IL-1β induced NO production by chondrocytes and cartilage. This effect appeared to result from iNOS transcriptional and/or post-transcriptional events, maybe by the inhibition …of the NF-κB transcription factor. This paper presents results on the influence of Diacerein on NO production. Show more

Keywords: Articular chondrocytes, diacerein, nitric oxide, NO synthase

Citation: Bio-Medical Materials and Engineering, vol. 18, no. s1, pp. 99-104, 2008

Authors: Féréol, S. | Fodil, R. | Laurent, V.M. | Planus, E. | Louis, B. | Pelle, G. | Isabey, D.

Article Type: Research Article

Abstract: The sensitivity of alveolar macrophages to substrate properties has been described in a recent paper (Féréol et al., Cell Motil. Cytoskel. 63 (2006), 321–340). It is presently re-analyzed in terms of F-actin structure (assessed from 3D-reconstructions in fixed cells) and mechanical properties (assessed by Magnetic Twisting Cytometry experiments in living cells) of cortical and deep cytoskeleton structures for rigid plastic (Young Modulus: 3 MPa) or glass (70 MPa) substrates and a soft (~0.1 kPa) confluent monolayer of alveolar epithelial cells. The cortical cytoskeleton component (lowest F-actin density) is represented by the rapid and softer viscoelastic compartment while the deep cytoskeleton …component (intermediate F-actin density) is represented by the slow and stiffer compartment. Stiffness of both cortical and deep cytoskeleton is significantly decreased when soft confluent monolayer of alveolar epithelial cells replace the rigid plastic substrate while F-actin reconstructions reveal a consistent actin cytoskeleton remodeling observable on both cytoskeleton components. Show more

Citation: Bio-Medical Materials and Engineering, vol. 18, no. s1, pp. 105-118, 2008