Bio-Medical Materials and Engineering - Volume 20, issue 3-4

Authors: Stoltz, J.-F. | Magdalou, J. | Netter, P. | Pinzano, A.

Article Type: Other

DOI: 10.3233/BME-2010-0622

Citation: Bio-Medical Materials and Engineering, vol. 20, no. 3-4, pp. 119-119, 2010

Authors: Brévignon-Dodin, Laure

Article Type: Research Article

Abstract: The EU Regulation on Advanced Therapy Medicinal Products (ATMP) bridges a regulatory gap and is expected to act as an enabler for the regenerative medicine sector in the EU by setting a centralised and harmonised regulatory framework for tissue-engineered products. Some of its key features are a workable and comprehensive scope, a new committee allowing for a pooling of expertise and tailored yet flexible requirements meant to keep pace with technology development. However, while providing a much needed regulatory framework, the new regulation still has potential shortfalls with regard to facilitating the research and commercialisation of tissue-engineered products in the …future. Show more

Keywords: ATMP Regulation, tissue-engineered products, flexible and anticipatory regulation, economic issues

DOI: 10.3233/BME-2010-0623

Citation: Bio-Medical Materials and Engineering, vol. 20, no. 3-4, pp. 121-126, 2010

Authors: de Isla, N. | Huseltein, C. | Jessel, N. | Pinzano, A. | Decot, V. | Magdalou, J. | Bensoussan, D. | Stoltz, J.-F.

Article Type: Research Article

Abstract: Tissue engineering is a multidisciplinary field that applies the principles of engineering, life sciences, cell and molecular biology toward the development of biological substitutes that restore, maintain, and improve tissue function. In Western Countries, tissues or cells management for clinical uses is a medical activity governed by different laws. Three general components are involved in tissue engineering: (1) reparative cells that can form a functional matrix; (2) an appropriate scaffold for transplantation and support; and (3) bioreactive molecules, such as cytokines and growth factors that will support and choreograph formation of the desired tissue. These three components may be used …individually or in combination to regenerate organs or tissues. Thus the growing development of tissue engineering needs to solve four main problems: cells, engineering development, grafting and safety studies. Show more

Keywords: Tissue engineering, cartilage, mechanobiology, mesenchymal stem cells

DOI: 10.3233/BME-2010-0624

Citation: Bio-Medical Materials and Engineering, vol. 20, no. 3-4, pp. 127-133, 2010

Authors: Cucchiarini, Magali | Madry, Henning

Article Type: Research Article

Abstract: Reproduction of a native, functional architecture in articular cartilage defects is a major problem in orthopaedic surgery. The elaboration of workable options to heal damaged cartilage might necessitate to involve cellular, molecular and environmental components to allow for the formation of an adequate and stable repair tissue in sites of injury. Strategies based on the transfer of candidate sequences to progenitor cells offer powerful tools to achieve this goal. The aim of this report is to provide an overview of the most recent therapeutic approaches developed in experimental orthopaedic research.

DOI: 10.3233/BME-2010-0625

Citation: Bio-Medical Materials and Engineering, vol. 20, no. 3-4, pp. 135-143, 2010

Authors: Ronzière, M.C. | Perrier, E. | Mallein-Gerin, F. | Freyria, A.-M.

Article Type: Research Article

Abstract: Regarding cartilage repair, tissue engineering is currently focusing on the use of adult mesenchymal stem cells (MSC) as an alternative to autologous chondrocytes. The potential of stem cells from various tissues to differentiate towards the chondrogenic phenotype has been investigated and it appears that the most common and studied sources are bone marrow (BM) and adipose tissue (AT) for historical and easy access reasons. In addition to three dimensional environment, the presence of member(s) of the transforming growth factor (TGF-β family and low oxygen tension have been reported to promote the in vitro differentiation of MSCs. Our work aimed at …characterizing and comparing the degree of chondrogenic differentiation of MSCs isolated from BM and AT cultured in the same conditions. We also further aimed at and at determining whether hypoxia (2% oxygen) could affect the chondrogenic potential of AT-MSCs. Cells were first expanded in the presence of FGF-2, then harvested and centrifuged to allow formation of cell pellets, which were cultured in the presence of TGF-β3 and/or Bone Morphogenetic Protein-2 (BMP-2) and with 2 or 20% oxygen tension, for 24 days. Markers of the chondrocyte (COL2A1, AGC1, Sox9) and hypertrophic chondrocyte (COL10A1, MMP-13) were monitored by real-time PCR and/or by immunohistological staining. Our data show that BMP-2/TGF-β3 combination is the best culture condition to induce the chondrocyte phenotype in pellet cultures of BM and AT-MSCs. Particularly, a switch in the expression of the pre-chondrogenic type IIA form to the cartilage-specific type IIB form of COL2A1 was observed. A parallel increase in gene expression of COL10A1 and MMP-13 was also recorded. However when AT-MSCs were cultured in hypoxia, the expression of markers of hypertrophic chondrocytes decreased when BMP-2/TGF-β3 were present in the medium. Thus it seems that hypoxia participates to the control of AT-MSCs chondrogenesis. Altogether, these cellular model systems will help us to investigate further the potential of different adult stem cells for cartilage engineering. Show more

Keywords: Bone marrow-derived stem cell, adipose tissue-derived stem cell, chondrogenesis, FGF-2, BMP-2, TGF-β3, hypoxia

DOI: 10.3233/BME-2010-0626

Citation: Bio-Medical Materials and Engineering, vol. 20, no. 3-4, pp. 145-158, 2010

Authors: Merceron, C. | Portron, S. | Masson, M. | Fellah, B.H. | Gauthier, O. | Lesoeur, J. | Chérel, Y. | Weiss, P. | Guicheux, J. | Vinatier, C.

Article Type: Research Article

Abstract: Articular cartilage does not repair itself spontaneously. To promote its repair, the transfer of stem cells from adipose tissue (ATSC) using an injectable self-setting cellulosic-hydrogel (Si-HPMC) appears promising. In this context, the objective of this work was to investigate the influence of in vitro chondrogenic differentiation of ATSC on the in vivo cartilage formation when combined with Si-HPMC. In a first set of experiments, we characterized ATSC for their ability to proliferate, self renew and express typical mesenchymal stem cell surface markers. Then, the potential of ATSC to differentiate towards the chondrogenic lineage and the optimal culture conditions to drive …this differentiation were evaluated. Real-time RT-PCR and histological analysis for sulphated glycosaminoglycans and type II collagen revealed that 3-dimensional culture and hypoxic condition favored ATSC chondrogenesis regarding mRNA expression level and the corresponding proteins production. In order to assess the phenotypic stability of chondrogenically-differentiated ATSC, real-time RT-PCR for specific terminal chondrogenic markers and alkaline phosphatase activity assay were performed. In addition to promote chondrogenesis, our culture conditions seem to prevent the terminal differentiation of ATSC. Histological examination of ATSC/Si-HPMC implants suggested that the in vitro chondrogenic pre-commitment of ATSC in monolayer is sufficient to obtain cartilaginous tissue in vivo. Show more

Keywords: Articular cartilage, tissue engineering, human mesenchymal stem cells, hydrogel, hypoxia

DOI: 10.3233/BME-2010-0627

Citation: Bio-Medical Materials and Engineering, vol. 20, no. 3-4, pp. 159-166, 2010

Authors: Tritz-Schiavi, J. | Charif, N. | Henrionnet, C. | de Isla, N. | Bensoussan, D. | Magdalou, J. | Benkirane-Jessel, N. | Stoltz, J.F. | Huselstein, C.

Article Type: Research Article

Abstract: Cartilage tissue engineering gives the ability to product adaptable neocartilage to lesion with autologous cells. Our work aimed to develop a stratified scaffold with a simple and progressive spraying build-up to mimic articular cartilage environment. An Alginate/Hyaluronic Acid (Alg/HA) hydrogel seeded with human Mesenchymal Stem Cells (hMSC) was construct by spray. First, cells repartition and actin organization were study with confocal microscopy. Then, we analyzed cells viability and finally, metabolic activity. Our results indicated a homogenous cells repartition in the hydrogel and a pericellular actin repartition. After 3 days of culture, we observed about 52% of viable cells in the …scaffold. Then, from day 7 until the end of culture (D28), the proportion of living cells and their metabolic activity increased, what indicates that culture conditions are not harmful for the cells. We report here that sprayed method allowed to product a scaffold with hMSCs that confer a favorable environment for neocartilage construction: 3D conformation and ability of cells to increase their metabolic activity, therefore with few impact on hMSCs. Show more

Keywords: Cartilage tissue engineering, sodium alginate, viability, metabolic activity, spraying cells

DOI: 10.3233/BME-2010-0628

Citation: Bio-Medical Materials and Engineering, vol. 20, no. 3-4, pp. 167-174, 2010

Authors: Henrionnet, Christel | Roeder, Emilie | Gillet, Romain | Galois, Laurent | Bensoussan, Danièle | Mainard, Didier | Netter, Patrick | Gillet, Pierre | Pinzano, Astrid

Article Type: Research Article

Abstract: This study investigated the gene expression profile of human mesenchymal stem cells seeded in collagen sponge for 28 days in three different mediums: (1) basal medium as control containing ITS alone, (2) ITS+TGF-β1 alone or (3) ITS 1% supplemented sequentially by TGF-β1 (D3–D14) followed by BMP-2 (D15–D28). Differential expression of 84 genes implicated in chondrogenic and osteogenic differentiation of MSCs was analyzed at D28 by real-time RT-PCR array technology. TGF-β1 alone down-regulated two genes, CD36 and cathepsin K. Sixteen genes were significantly up-regulated, notably type 2 and type 10 collagens, COMP and Sox9. The sequential combination of TGF-β1 and BMP-2 …produced a similar profile with prominent expression of type 2 collagen and the alkaline phosphatase gene. Interestingly, in this in vitro condition, RUNX2 was not up-regulated, suggesting that the sequential combination of TGF-β1/BMP2 enhances the hypertrophic chondrogenic profile without turning towards the osteoblastic pathway. Show more

Keywords: RT-PCR array, human mesenchymal stem cells, collagen sponge, gene expression, growth factors

DOI: 10.3233/BME-2010-0629

Citation: Bio-Medical Materials and Engineering, vol. 20, no. 3-4, pp. 175-181, 2010

Authors: Dumas, D. | Henrionnet, C. | Hupont, S. | Werkmeister, E. | Stoltz, J.F. | Pinzano, A. | Gillet, P.

Article Type: Research Article

Abstract: We propose an innovative invasiveless technique in the field of nonlinear optical imaging to facilitate monitoring of cell/scaffold combinations for tissue repair. By using a near infrared (NIR) femtosecond excitation, we were able to introduce a new index based on decay time response for fluorescence (F) and Second Harmonic Generation (SHG) obtained with Time Correlated Single Photon Counting (TCSPC) microscopy to monitor structural information on the state of the matrix collagen. Some human Mesenchymal Stem Cells (hMSCs) seeded in 3D scaffolds were tested with different culture times (from D7 to D56) to analyze the effect of Tumor Growth Factor beta …1 (TGF-β1) on type-2 collagen expression in the matrix. After 14 days in the presence of TGF-β1, our results showed an increase in the expression of type-2 collagen synthesized by hMSCs, and a change in collagen conformation, as an indication of its ability to be detected as a harmonophore by TCSPC–SHG without the need for an exogenous probe. Show more

Keywords: Tissue engineering, lifetime, second harmonic microscopy, stem cells, collagen, extracellular matrix

DOI: 10.3233/BME-2010-0630

Citation: Bio-Medical Materials and Engineering, vol. 20, no. 3-4, pp. 183-188, 2010

Authors: Goebel, Jean-Christophe | Pinzano, Astrid | Grenier, Denis | Perrier, Anne-Laure | Henrionnet, Christel | Galois, Laurent | Gillet, Pierre | Beuf, Olivier

Article Type: Research Article

Abstract: Due to the actual interest for bioengineering in the osteoarthritis (OA) healing context, researchers need accurate qualitative and quantitative methodologies to evaluate in vivo the integration and functionality of their cartilage-like biomaterials. As in clinical diagnostic strategies, advances in Magnetic Resonance Imaging (MRI) seem promising for non-vulnerant assessments of articular cartilage bio-architecture and morphology in small animal models. These experimental models are commonly used to monitor the physiopathology of OA and to evaluate therapeutic responses mediated by chondroprotective drugs or tissue engineering. Nowadays, the application of MR protocols to in vivo small animal cartilage imaging is achievable with the development …of high magnetic fields and the adaptation of methodologies to reach the required spatial resolution and contrast. The purpose of this article is to summarize these current MRI strategies used for in vivo small animal articular cartilage assessments. Show more

Keywords: MRI, cartilage, osteoarthritis, animal models, bioengineering, extracellular matrix

DOI: 10.3233/BME-2010-0631

Citation: Bio-Medical Materials and Engineering, vol. 20, no. 3-4, pp. 189-194, 2010