Bio-Medical Materials and Engineering - Volume 23, issue 4

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

Article Type: Other

DOI: 10.3233/BME-130748

Citation: Bio-Medical Materials and Engineering, vol. 23, no. 4, pp. 249-249, 2013

Authors: Beroud, J. | Vincent, B. | Paternotte, E. | Nguyen, V.-S. | Kerdjoudj, H. | Velot, E. | Rouxel, D. | Menu, P.

Article Type: Research Article

Abstract: BACKGROUND: In tissue engineering, the endothelialization of vascular scaffold can be a crucial step to improve graft patency. A functional cellularization requires coating surfaces. Since 2003, our group used polyelectrolyte multilayer films (PEMFs) made of poly(allylamine hydrochloride) and polystyren sulfonate to coat luminal surface of blood vessel. Previous results showed that PEMFs have remarkable effect on cellular behavior: adhesion, proliferation, differentiation. However, no method seems adapted for in vitro measurement of the viscoelastic shift after PEMFs buildup. OBJECTIVE: In this present work, we proposed to use a new analytical method based on Brillouin spectroscopy (BS) to investigate the influence …PEMFs coating on vessel intrinsic viscoelasticy. METHODS: On human umbilical arteries and rabbit vessels, PEMFs were buildup and the luminal surfaces viscoelasticy were measuring by BS. RESULTS: It seems that these films do not alter dynamic functionality and BS could be an interesting method for understanding the role of the tissue architecture, the interrelation between the different structures constituting the wall and the influence of this architecture on the tissue behavior, especially with the characterized components of the different vascular wall. CONCLUSION: The ability of BS to characterize biological samples opens potential applications in tissue engineering field, especially as a tool for a better understanding of vascular diseases. Show more

Keywords: Polyelectrolyte multilayer films, Brillouin spectroscopy, viscoelasticity, vascular substitutes

DOI: 10.3233/BME-130749

Citation: Bio-Medical Materials and Engineering, vol. 23, no. 4, pp. 251-261, 2013

Authors: Chen, Bailiang | Roeder, Emilie | Vuissoz, Pierre-André | Gillet, Pierre | Felblinger, Jacques | Beaumont, Marine | Pinzano, Astrid

Article Type: Research Article

Abstract: Non-invasive quantitative assessment of articular cartilage integrity is essential for early detection and evaluation of osteoarthritis (OA) and for the follow-up of stem-cell-driven cartilage engineering. In this study, we investigated the feasibility of exploiting diffusion tensor imaging (DTI) on porcine knee joints with a clinical magnetic resonance (MR) scanner to extract micro-structural information in order to complement biochemical information quantified by T2 maps. We propose an MR protocol for quantifying T2 and cartilage microstructure with diffusion MR on a clinical scanner. Preliminary results were obtained on four pig knee joints using a 3 T GE clinical MRI scanner and an …8-channel knee coil array. The measured cartilage volume, T2 values, apparent diffusion coefficient and fractional anisotropy (FA) of femoral and tibial cartilage were respectively 9.8/2.3 mm2 , 67.0/56.1 ms, 1.3/1.3×10−3 mm2 /s and 0.4/0.3. This new protocol has the potential to be combined in vivo with quantitative assessment of both cartilage degradation and restoration in osteoarthritis. Show more

Keywords: Osteoarthritis, stem cell therapy, quantitative assessment, MR, diffusion tensor imaging

DOI: 10.3233/BME-130750

Citation: Bio-Medical Materials and Engineering, vol. 23, no. 4, pp. 263-272, 2013

Authors: Harmouch, C. | El-Omar, R. | Labrude, P. | Decot, V. | Menu, P. | Kerdjoudj, H.

Article Type: Research Article

Abstract: BACKGROUND: Mesenchymal stem cells (MSCs) are multipotent cells able to differentiate into several lineages with valuable applications in regenerative medicine. MSCs differentiation is highly dependent on physicochemical properties of the culture substrate, cell density and on culture medium composition. OBJECTIVE: In this study, we assessed the influence of fetal bovine serum (FBS) level on Wharton's jelly (WJ)-MSCs behavior seeded on polyelectrolyte multilayer films (PEMF) made of four bilayers of poly-allylamine hydrochloride (PAH) as polycation and poly-styrene sulfonate (PSS) as polyanion. METHODS: MSCs isolated from WJ by explants method were amplified until the third passage. Their phenotypic characterization was …performed by flow cytometry analyses. MSCs were seeded on PEMF, in Endothelial growth medium-2 (EGM-2) supplemented by either 5% or 2% FBS. Cell's behavior was monitored for 20 days by optical microscopy and immunofluorescence. RESULTS: Until 2 weeks on glass slides, no difference was observed whatever the FBS percentage. Then with 5% FBS, MSCs formed three-dimensional spheroids on PSS/PAH after 20 days of culture with a nuclear aggregate. Whereas, with 2% FBS, these spheroids did not appear and cells grown in 2D conserved the fibroblast-like morphology. CONCLUSIONS: The decrease of FBS percentage from 5% to 2% avoids 3D cell spheroids formation on PAH/PSS. Such results could guide bioengineering towards building 2D structures like cell layers or 3D structures by increasing the osteogenic or chondrogenic differentiation potential of MSCs. Show more

Keywords: MSCs, fetal bovine serum, spheroids, tissue engineering

DOI: 10.3233/BME-130751

Citation: Bio-Medical Materials and Engineering, vol. 23, no. 4, pp. 273-280, 2013

Authors: Huot, Stéphane | Rohman, Géraldine | Riffault, Mathieu | Pinzano, Astrid | Grossin, Laurent | Migonney, Véronique

Article Type: Research Article

Abstract: BACKGROUND: Biodegradable polymers used in tissue engineering applications, such as poly(ε-caprolactone) (PCL), are hydrophobic leading to a lack of favorable cell signalization and finally to a poor cell adhesion, proliferation and differentiation. To overcome this problem, scaffolds undergo generally a surface modification. OBJECTIVE: Our laboratory has demonstrated that the grafting of poly(sodium styrene sulfonate) (pNaSS) onto titanium or poly(ethylene terephthalate) surfaces, leads to a more specific protein adsorption and a better control of cell proliferation. The objective of this work is to develop, through a straightforward way, bioactive elastomeric PCL scaffolds by grafting pNaSS. METHODS: Porous elastomeric PCL …scaffolds were developed using a particulate-leaching process. pNaSS was grafted into the scaffold by a “grafting from” technique. In vitro tests were carried out to assess cell adhesion and protein expression. RESULTS: pNaSS was grafted homogeneously onto PCL scaffolds without degrading the biodegradable polymer or the porous structure. The in vitro studies have shown that pNaSS grafted onto PCL improves the cell response with a better expression of collagen, fibronectin and integrin α1. CONCLUSIONS: The grafting of pNaSS onto biomaterial surfaces is a versatile method that can provide a new generation of biodegradable scaffolds which could be “biointegrable”. Show more

Keywords: Tissue engineering, porous scaffolds, bioactive polymer, elastomeric PCL, pNaSS

DOI: 10.3233/BME-130752

Citation: Bio-Medical Materials and Engineering, vol. 23, no. 4, pp. 281-288, 2013

Authors: Lessim, Soucounda | Migonney, Véronique | Thoreux, Patricia | Lutomski, Didier | Changotade, Sylvie

Article Type: Research Article

Abstract: BACKGROUND: Introduction of a new generation of artificial ligaments for ACL reconstruction, the Ligament Augmentation and Reconstruction System (LARS), gives promising clinical results [1]. The current literature supports the use of LARS from short to medium term. To go even further to improve the biocompatibility of this biomaterial, poly(sodium styrene sulfonate) (polyNaSS) was grafted onto its surface. Studies using sheep animal model showed improvement of knee functionalities with this grafted artificial ligament and a better adhesion of human cell lines. OBJECTIVES: To better understand this in vivo improvement of integration with the bioactivated artificial prosthesis, in vitro studies were …leaded using human ligament fibroblasts. METHODS: Human ligament fibroblasts isolated from human ruptured ACL were amplified and seeded onto poly(NaSS) grafted and non-grafted PET scaffold (Lars ligament) under standard culture conditions. Cellularized fibers were observed under scanning electron microscopy and histological and immunohistological studies were performed. RESULTS: Cells are localized around the grafted PET fibers of the bioactive ligament and penetrate in the scaffold. On ungrafted fibers, cells stay around the scaffold. On grafted fibers, collagen I appears strongly organized whereas is thin and dispersed on non grafted fibers. Finally, grafting altered localization of decorin. CONCLUSIONS: PolyNaSS grafting enhances human ligament fibroblast organisation in vitro in contact with biomaterial and improves collagen and decorin deposits around fibers. Show more

Keywords: Anterior cruciate ligament (ACL), fibroblast, matrix, biointegration, biomaterial, bioactive surface

DOI: 10.3233/BME-130753

Citation: Bio-Medical Materials and Engineering, vol. 23, no. 4, pp. 289-297, 2013

Authors: Rammal, H. | Beroud, J. | Gentils, M. | Labrude, P. | Menu, P. | Kerdjoudj, H. | Velot, E.

Article Type: Research Article

Abstract: BACKGROUND: Polyelectrolyte multilayer (PEMs) films made of poly(allylamine hydrochloride) (PAH) as polycation and poly(styrene sulfonate) (PSS) as polyanion, with a PAH ending layer, can be used as a coating in order to improve the anti-thrombogenicity and patency of vascular grafts in vascular engineering field. They induce strong adhesion of mature endothelial cells on glass, expanded polytetrafluoroethylene and cryopreserved arteries. Despite their outstanding effect on mature and progenitor endothelial cells, PEMs ending with PAH showed a poor outcome on Wharton's jelly mesenchymal stem cells (WJ-MSCs) culture. OBJECTIVE: The aim of this work was to examine the influence of the ending …charge of PEMs on WJ-MSCs behavior. METHODS: WJ-MSCs amplified until the 3rd passage were seeded and cultured on (PAH-PSS)3 -PAH and on (PAH-PSS)4 coated glass for 10 days. Stem cell phenotype was checked by flow cytometry and cell morphology was followed by bright field microscopy. RESULTS: Flow cytometry analysis showed that WJ-MSCs were positive for MSC's markers CD73, CD90 and CD105 and negative for hematopoietic markers CD34 and CD45. Light microscopy showed development of nodule-like structures after 10 days of culture on (PAH-PSS)3 -PAH, which resulted in a disturbance of cell monolayer. Whereas WJ-MSCs cultured on (PAH-PSS)4 ending with PSS showed a normal cell growth like on collagen and reached confluence after 10 days. CONCLUSION: The culture surface seems to have a determining role in WJ-MSC's “spatial” behavior, which could be considered in the field of tissue engineering. Show more

Keywords: Mesenchymal stem cells, polyelectrolyte multilayer films, culture surface, stem cells behavior

DOI: 10.3233/BME-130754

Citation: Bio-Medical Materials and Engineering, vol. 23, no. 4, pp. 299-309, 2013

Authors: Laydi, F. | Rahouadj, R. | Cauchois, G. | Stoltz, J.-F. | de Isla, N.

Article Type: Research Article

Abstract: Collagen gels could be used as carriers in tissue engineering to improve cell retention and distribution in the defect. In other respect hydroxyapatite could be added to gels to improve mechanical properties and regulate gel contraction. The aim of this work was to analyze the feasibility to incorporate hydroxyapatite into collagen gels and culture mesenchymal stem cells inside it. Human bone marrow mesenchymal stem cells (hMSC-BM) were used in this study. Gels were prepared by mixing rat tail type I collagen, hydroxyapatite microparticles and MSCs. After polymerization gels were kept in culture while gel contraction and mechanical properties were studied. …In parallel, cell viability and morphology were analyzed. Gels became free-floating gels contracted from day 3, only in the presence of cells. A linear rapid contraction phase was observed until day 7, then a very slow contraction phase took place. The incorporation of hydroxyapatite improved gel stability and mechanical properties. Cells were randomly distributed on the gel and a few dead cells were observed all over the experiment. This study shows the feasibility and biocompatibility of hydroxyapatite supplemented collagen gels for the culture of mesenchymal stem cells that could be used as scaffolds for cell delivery in osteoarticular regenerative medicine. Show more

Keywords: Collagen gel, hydroxyapatite, mesenchymal stem cells, tissue engineering

DOI: 10.3233/BME-130755

Citation: Bio-Medical Materials and Engineering, vol. 23, no. 4, pp. 311-315, 2013