Bio-Medical Materials and Engineering - Volume 25, issue s1

Authors: Stoltz, Jean François | Zhang, Lei

Article Type: Other

DOI: 10.3233/BME-141224

Citation: Bio-Medical Materials and Engineering, vol. 25, no. s1, pp. 1-2, 2015

Authors: Stoltz, J.-F. | Bensoussan, D. | Zhang, L. | Decot, V. | De Isla, N. | Li, Y.P. | Huselstein, C. | Benkirane-Jessel, N. | Li, N. | Reppel, L. | He, Y. | Li, Y.Y.

Article Type: Review Article

Abstract: Since the 1960s and the therapeutic use of hematopoietic stem cells of bone marrow origin, there has been increasing interest in the study of undifferentiated progenitors that have ability to proliferate and differentiate in different tissues. Different stem cells (SC) with different potential can be isolated and characterised. Despite the promise of embryonic stem cells, in many cases, adult stem cells provide a more interesting approach to clinical applications. It is undeniable that mesenchymal stem cells (MSC) from bone marrow, adipose tissue or MSC of Wharton Jelly, which have limited potential, are of interest for clinical applications in regenerative medicine …because they are easily separated and prepared and no ethical problems are involved in their use. During the last 10 years, these multipotent cells have generated considerable interest and in particular have been shown to escape allogeneic immune response and be capable of immunomodulatory activity. These properties may be of a great interest for regenerative medicine. Different clinical applications are under study (cardiac insufficiency, atherosclerosis, stroke, bone, cartilage, diabetes, ophthalmology, urology, liver, organ's reconstruction…). Show more

Keywords: Stem cells, tissue engineering, clinical studies, international regulations, forecast markets

DOI: 10.3233/BME-141225

Citation: Bio-Medical Materials and Engineering, vol. 25, no. s1, pp. 3-26, 2015

Authors: Massin, Frédéric | Huili, Cai | Decot, Véronique | Stoltz, Jean-François | Bensoussan, Danièle | Latger-Cannard, Véronique

Article Type: Research Article

Abstract: INTRODUCTION: Stem cells for autologous and allogenic transplantation are obtained from several sources including bone marrow, peripheral blood or cord blood. Accurate enumeration of viable CD34+ hematopoietic stem cells (HSC) is routinely used in clinical settings, especially to monitor progenitor cell mobilization and apheresis. The number of viable CD34+ HSC has also been shown to be the most critical factor in haematopoietic engraftment. The International Society for Cellular Therapy actually recommends the use of single-platform flow cytometry system using 7-AAD as a viability dye. AIM: In a way to move routine analysis from a BD FACSCaliburTM instrument to …a BD FACSCantoTM II, according to ISO 15189 standard guidelines, we define laboratory performance data of the BDTM Stem Cell Enumeration (SCE) kit on a CE-IVD system including a BD FACSCanto II flow cytometer and the BD FACSCantoTM Clinical Software. InterQCTM software, a real time internet laboratory QC management system developed by VitroTM and distributed by Becton DickinsonTM , was also tested to monitor daily QC data, to define the internal laboratory statistics and to compare them to external laboratories. METHODS: Precision was evaluated with BDTM Stem Cell Control (high and low) results and the InterQC software, an internet laboratory QC management system by Vitro. This last one drew Levey–Jennings curves and generated numeral statistical parameters allowing detection of potential changes in the system performances as well as interlaboratory comparisons. Repeatability, linearity and lower limits of detection were obtained with routine samples from different origins. Agreement evaluation between BD FACSCanto II system versus BD FACSCalibur system was tested on fresh peripheral blood, freeze-thawed apheresis, fresh bone marrow and fresh cord blood samples. RESULTS: Instrument's measure and staining repeatability clearly evidenced acceptable variability on the different samples tested. Intra- and inter-laboratory CV in CD34+ cell absolute count are consistent and reproducible. Linearity analysis, established between 2 and 329 cells/μl showed a linear relation between expected counts and measured counts (R2 =0.97). Linear regression and Bland–Altman representations showed an excellent correlation on samples from different sources between the two systems and allowed the transfer of routine analysis from BD FACSCalibur to BD FACSCanto II. CONCLUSIONS: The BD SCE kit provides an accurate measure of the CD34 HSC, and can be used in daily routine to optimize the enumeration of hematopoietic CD34+ stem cells by flow cytometry. Moreover, the InterQC system seems to be a very useful tool for laboratory daily quality monitoring and thus for accreditation. Show more

Keywords: CD34+ hematopoietic stem cells, flow cytometry, hematopoietic transplantation

DOI: 10.3233/BME-141226

Citation: Bio-Medical Materials and Engineering, vol. 25, no. s1, pp. 27-39, 2015

Authors: Benameur, Laila | Charif, Naceur | Li, Yueying | Stoltz, Jean-François | de Isla, Natalia

Article Type: Research Article

Abstract: Under physiological conditions, there is a production of limited range of free radicals. However, when the cellular antioxidant defence systems, overwhelm and fail to reverse back the free radicals to their normal basal levels, there is a creation of a condition of redox disequilibrium termed “oxidative stress”, which is implicated in a very wide spectrum of genetic, metabolic, and cellular responses. The excess of free radicals can, cause unfavourable molecular alterations to biomolecules through oxidation of lipids, proteins, RNA and DNA, that can in turn lead to mutagenesis, carcinogenesis, and aging. Mesenchymal stem cells (MSCs) have been proven to be …a promising source of cells for regenerative medicine, and to be useful in the treatment of pathologies in which tissue damage is linked to oxidative stress. Moreover, MSCs appeared to efficiently manage oxidative stress and to be more resistant to oxidative insult than normal somatic cells, making them an interesting and testable model for the role of oxidative stress in the aging process. In addition, aging is accompanied by a progressive decline in stem cell function, resulting in less effective tissue homeostasis and repair. Also, there is an obvious link between intracellular reactive oxygen species levels and cellular senescence. To date, few studies have investigated the promotion of aging by oxidative stress on human MSCs, and the mechanism by which oxidative stress induce stem cell aging is poorly understood. In this context, the aim of this review is to gain insight the current knowledge about the molecular mechanisms of aging-induced oxidative stress in human MSCs. Show more

Keywords: Aging, oxidative stress, human mesenchymal stem cells

DOI: 10.3233/BME-141247

Citation: Bio-Medical Materials and Engineering, vol. 25, no. s1, pp. 41-46, 2015

Authors: Liu, Xing | Xie, Yi | Li, Wei | Sheng, Wen | Li, Yinping | Tong, Zan | Ni, Hong | Huselstein, Celine | Wang, Xiong | Chen, Yun

Article Type: Research Article

Abstract: A series of composite films were prepared from glycerol-plasticized starch and zein by intensive mixing and hot press. The structure and physical properties of the starch/zein (SZ) composite films were characterized by scanning electron microscope (SEM), optical microscopy and water contract angle testing. The hemocompatibility and cytocompatibility of SZ films were evaluated by plasma recalcification time, hemolysis assay and cell culture experiment. SEM and optical observation showed that starch and zein domains can be differed in the films and in a two phase separation status. Glycerol affects the surface hydrophilicity/hydrophobicity of the films. The hemocompatibility and cytocompatibility evaluation showed that …SZ composites are anticoagulant materials with no hemolysis and low cytotoxicity. The SZ composites maybe have potentials for applications as biomaterials. Show more

Keywords: Starch, zein, hot press, biomaterials

DOI: 10.3233/BME-141227

Citation: Bio-Medical Materials and Engineering, vol. 25, no. s1, pp. 47-55, 2015

Authors: Luo, Lihua | Gong, Wenrong | Zhou, Yi | Yang, Lin | Li, Daokun | Huselstein, Celine | Wang, Xiong | He, Xiaohua | Li, Yinping | Chen, Yun

Article Type: Research Article

Abstract: OBJECTIVE: To evaluate the in vitro cytocompatibility of cellulose/soy protein isolate composite membranes (CSM) with Schwann cells and in vivo toxicity to animals. METHODS: A series of cellulose/soy protein isolate composite membranes (CSM) were prepared by blending, solution casting and coagulation process. The cytocompatibility of the CSM to Schwann cells were evaluated by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and by direct cells culture of Schwann cells on the surfaces of the CSM, respectively. The in vivo toxicity of the CSM to animals were also evaluated by acute toxicity testing, skin sensitization testing, pyrogen testing and intracutaneous stimulation testing, respectively, according …to the ISO 10993 standard. RESULTS: The MTT assay showed that the cell viability of Schwann cells cultured in extracts from the CSM was higher than that from the neat cellulose membrane without containing SPI component. The direct cells culture indicated that the Schwann cells could attach and grow well on the surface of the CSM and the incorporation of SPI into cellulose contributed to improvement of cell adhesion and proliferation. The evaluations of in vivo biological safety suggested that the CSM showed no acute toxicity, no skin sensitization and no intracutaneous stimulation to the experimental animals. CONCLUSION: The CSM had in vitro cytocompatibility with Schwann cells and biological safety to animals, suggesting potential for the applications as nerve conduit for the repair of nerve defect. Show more

Keywords: Cellulose, soy protein isolate, nerve conduit, cytocompatibility, acute toxicity

DOI: 10.3233/BME-141228

Citation: Bio-Medical Materials and Engineering, vol. 25, no. s1, pp. 57-64, 2015

Authors: Xin, Yi | Wu, Guanghui | Wu, Man | Zhang, Xiaoxia | Velot, Emilie | Decot, Véronique | Cui, Wei | Huang, Yimin | Stoltz, Jean-Francois | Du, Jie | Li, Na

Article Type: Research Article

Abstract: The scaffolds prepared from the tissue decellularization conserve the porous 3-D structure and provide an optimal matrix for the tissue regeneration. Since decade, the enzymatic digestion, chemical reagent treatment and mechanical actions such as eversion and abrasion have been used to remove the cells from the intact matrix. In this study, we optimized an enzymatic method to decellularize the umbilical artery to construct a 3-D porous scaffold which is suitable for the culture of mesenchymal stem cells (MSCs). The scaffold maintained the interconnected porous structure. It remained the similar high water content 95.3±1% compared to 94.9±0.6% in the intact umbilical …artery (p>0.05). The decellularization process decreased the stress from 0.24±0.05 mPa to 0.15±0.06 mPa (p<0.05). However the decellularization did not change the strain of the artery (45±15% vs. 53±10%, p>0.05). When the scaffold was transplanted to the subcutaneous tissue in the wild type mice, there were less T cells appeared in the surrounding tissue which meant the decreased the immunogenicity by decellularization. This scaffold also supported the adhesion and proliferation of the MSCs. In this study, we constructed a biological compatible porous scaffold from the decellularized umbilical artery which may provide a suitable scaffold for cell-matrix interaction studies and for tissue engineering. Show more

Keywords: Decellularization, biological scaffold, umbilical artery, tissue engineering, mesenchymal stem cells

DOI: 10.3233/BME-141261

Citation: Bio-Medical Materials and Engineering, vol. 25, no. s1, pp. 65-71, 2015

Authors: Lehalle, Bertrand

Article Type: Research Article

Abstract: Because many of patients with peripheral arterial disease are not eligible for direct or conventional revascularization procedures, because stem cell therapy is being investigated as to its possible role in the treatment of limb ischemia, there is a need to evaluate this treatment and his true application. On the basis of experimental data, preliminary clinical studies have established the safety and feasibility of stem cells implantation in case of critical limb ischemia. Forthcoming large studies, especially randomized placebo controlled double blind studies, related to the optimal cell type, dosage, administration route, will consolidate this evidence and establish mid and long …term effectiveness. Show more

Keywords: Cell therapy, peripheral artery disease, critical limb ischemia, mesenchymal stem cells

DOI: 10.3233/BME-141229

Citation: Bio-Medical Materials and Engineering, vol. 25, no. s1, pp. 73-78, 2015

Authors: Eap, Sandy | Morand, David | Clauss, François | Huck, Olivier | Stoltz, Jean-François | Lutz, Jean-Christophe | Gottenberg, Jacques-Eric | Benkirane-Jessel, Nadia | Keller, Laetitia | Fioretti, Florence

Article Type: Research Article

Abstract: Designing unique nanostructured biomimetic materials is a new challenge in modern regenerative medicine. In order to develop functional substitutes for damaged organs or tissues, several methods have been used to create implants able to regenerate robust and durable bone. Electrospinning produces nonwoven scaffolds based on polymer nanofibers mimicking the fibrillar organization of bone extracellular matrix. Here, we describe a biomimetic 3D thick nanofibrous scaffold obtained by electrospinning of the biodegradable, bioresorbable and FDA-approved polymer, poly(ε-caprolactone). Such scaffold presents a thickness reaching one centimeter. We report here the demonstration that the designed nanostructured implant is able to induce in vivo bone …regeneration. Show more

Keywords: Bone induction, tissue engineering, 3D scaffold, polycaprolactone, electrospinning

DOI: 10.3233/BME-141248

Citation: Bio-Medical Materials and Engineering, vol. 25, no. s1, pp. 79-85, 2015

Authors: Filip, Anna | Bianchi, Arnaud | Mainard, Didier | Lacolley, Patrick | Magdalou, Jacques | Mercier, Nathalie

Article Type: Research Article

Abstract: OBJECTIVES: Chondrocytes hypertrophy is a physiological process observed in endochondral ossification during development until adolescence in human. It can also be observed during pathophysiological conditions such as osteoarthritis. Hypertrophic chondrocytes synthesise collagen X and express matrix metalloproteinase 13 and alkaline phosphatase. The cellular models available to study this process are either not convenient, they might lead to a rapid dedifferentiation of chondrocytes, or they are far from the physiological conditions. The objective of this study was to design an user-friendly 2D-primary cell culture of young articular chondrocytes of rat able to follow the terminal differentiation process. EXPERIMENTAL DESIGN: After …confluence, chondrocytes were cultured according to 4 differentiation protocols. Protocol 1 contained DMEM/F12 supplemented with 10% foetal bovine serum (FBS) and 2 μg/ml insulin. Protocol 2 contained alpha-MEM supplemented with 5% FBS and 2 μg/ml insulin. Protocol 3 contained 2% FBS and 2 μg/ml insulin. Protocol 4 contained DMEM/F12 supplemented with 2% FBS in absence or in presence of 2 μg/ml insulin and 37.5 μg/ml ascorbate. The cell morphology was observed by phase-contrast microscopy and the expression of markers specific of mature and hypertrophic chondrocytes were assessed by RT-qPCR. RESULTS: The effect of a decrease in nutrient quality of the culture medium after confluence was tested using protocols 1, 2 and 3. Protocol 1 did not allow the maintenance of chondrocyte phenotype more than one week, because cells became fibroblastic. A decrease in Sox9 mRNA expression, in collagen II/collagen I and in aggrecan/versican mRNA ratios was also found with protocol 1. Protocol 3 was the best when compared with protocols 1 and 2. It allowed chondrocytes to adopt a hypertrophic morphology. Cells also expressed the collagen X specific hypertrophic marker, and presented an increase in collagen II/I and aggrecan/versican ratios after 15 days of culture post-confluence. The effect of the insulin/ascorbate supplementation was studied using protocol 4. The insulin/ascorbate supplementation allowed an earlier chondrocytes conversion to terminal differentiation with a prolonged effect till 3 weeks post-confluence, compared to control without insulin/ascorbate. Finally, the profile of chondrocyte differentiation was checked during 5 successive sub-cultures. Only the first passage could be used to study hypertrophy. CONCLUSION: A convenient protocol to study chondrocyte hypertrophy is proposed. Protocol 4 offers the possibility to study this differentiation phenotype which is crucial for the development of articular diseases such as osteoarthritis. Our model could also be used in tissue engineering for cartilage repair strategies in which hypertrophic differentiation of chondrocyte should be avoided. Show more

Keywords: Chondrocytes, articular cartilage, differentiation, hypertrophy, osteoarthritis

DOI: 10.3233/BME-141252

Citation: Bio-Medical Materials and Engineering, vol. 25, no. s1, pp. 87-102, 2015