- Email: [email protected]
salt: effect of the surface charge on precipitation
Abstracts / Current Opinion in Biotechnology 22S (2011) S15–S152
linear finite element method (FEM). First, the ankle model (tibia–fibia–calcaneus–talus–trabecular–cortical bone and soft tissue) was developed with computer tomography, then the nonlinear FEM was applied to analyze the stresses and strains occurred in ligaments and bones for four different positions (10◦ Dorsiflexion, 0◦ Neutralposition, 10◦ Plantarflexion, 20◦ Plantarflexion). The ligaments were attached to the relevant points according to the anatomical structure of foot. The soft tissue was defined as hyper elastic while other tissues were idealized as homogeneous, isotropic and linearly elastic. Anterior displacement of foot from high to down in terms of different positions was found to be as 0◦ N > 20◦ PF > 10◦ DF > 10◦ PF respectively by applying 80 N force on calcaneus in horizontal direction. The maximum displacements were found on calcaneus where the loads were applied and the minumum displacements were found on fibula and tibia where the supports were located. The finite element method can be used as a powerful tool in the simulation of living structures. Using of noninvasive methods such as FEM can help to determine behaviour of ankle and ligaments before the treatment (ADT) applied to the patients. doi:10.1016/j.copbio.2011.05.502
R3 Plasmid DNA precipitation in the presence of cationic surfactant/salt: effect of the surface charge on precipitation Nese Ataci 1 , Ayfer Sarac 2 , Inci Arisan 1 1
Department of Biochemistry, Yildiz Technical University, Istanbul, Turkey 2 Department of Physical Chemistry, Yildiz Technical University, Istanbul, Turkey E-mail address: [email protected] (N. Ataci) DNA is a biological material, a kind of anionic polymer with double helical structure. Surfactants are an active field of research in colloidal science; especially consist of ionic or non-ionic head group and long alkyl chains. Many investigations have been presented on surfactant/DNA complex formation, precipitation and microstructure by a variety of methods. Knowledge of the interaction between DNA with cationic surfactants has many applications; the development of methods of DNA extraction and purification and lately, the potential use of these systems as vehicles for gene delivery and gene transfection. In this study, we investigated the effect of N,N ,N -polyethylene (10)-N tallow–1,3-di amine propane (PEG–10 tallow)/salt on plasmid DNA (pDNA) precipitation in critical micelle concentration (CMC). In the first step, CMC of PEG-10 tallow was determined by surface tension, and conductometry methods, and it was obtained closely agree with those determined both methods. The salt effect on precipitation profiles of PEG-10 tallow/pDNA formation was examined with agarose gel electrophoresis. Surface charges of PEG-10 tallow surfactant in different concentrations and surfactant/salt solutions were determined by zeta potential measurement. Results showed that, increased ionic strength changed parallel with DNA precipitation profiles. doi:10.1016/j.copbio.2011.05.503
S151
R4 Relative gene expression analysis shows increased mRNA levels of HoxB7 and HoxA10 and decreased mRNA levels of Pbx1 in the bone marrow derived mesenchymal progenitors from Fanconi anemia patients Aysen Gunel Ozcan 1 , Ilgin Cagnan 1 , Baris Kuskonmaz 2 , Duygu Uckan Cetinkaya 1 1
Hacettepe University, Institute of Pediatry & Center for Stem Cell Research and Development, Ankara, Turkey 2 Hacettepe University, Pediatric Hematology Bone Marrow Transplantation Unit, Ankara, Turkey E-mail address: [email protected] (A.G. Ozcan) Of having important roles on anterio-posterior body axis determination, embryonic body formation, hematopoeisis, eye development and determination of the cell type identity or cellular proliferation and differentiation during neuronal specification, Hox and the three-amino-acid loop extension family (TALE) proteins are the center of various biomedical research. Several research data shows that Hox gene family has great impact on regulating the genes and establishment of the balance between cellular proliferation and differentiation. Our research group aims to understand the Hox/TALE network and its regulators in Fanconi anemia, a cancer predisposition disorder characterized by progressive bone marrow failure. In this study, bone marrow derived mesenchymal stromal cells from three Fanconi anemia patients were expanded and characterized for their adipogenic and osteogenic differentiation. Spontaneous adipogenic differentiation was observed in one of the patients’ samples. Mesenchymal stromal cells of two patients did not differentiate to osteogenic lineage in osteoblast induction medium whereas donor cells did. Relative expression of Hox and Pbx1 (from TALE family) genes in mesenchymal stromal cells of patients and donors was determined by real-time RT-PCR. Analysis, performed by 2-((Ct method, showed that HoxB7 and HoxA10 mRNA levels in the patients’ cells increased whereas Pbx1 mRNA levels decreased. doi:10.1016/j.copbio.2011.05.513
R5 Strategies combining cells and scaffolds for bone tissue engineering Enrica Saino 1 , Nora Imbriani 4 , Livia Visai 5
Bloise 2 , Lorenzo
Fassina 3 , Marcello
1
Center for Tissue Engineering (C.I.T), Pavia, Italy Dep. Biochemistry, University of Pavia, Italy 3 Dep. Computer and Systems Science, University of Pavia, Italy 4 Salvatore Maugeri Foundation IRCCS, Pavia, Italy 5 International Centre for Studies and Research in Biomedicine (ICB), Luxembourg 2
E-mail address: [email protected] (E. Saino) Engineering bone uses porous scaffolds, osteoblasts or cells that can become osteoblasts, and factors that promote cell attachment, differentiation, and mineralized bone formation. In this study we investigated the effects of the electromagnetic stimulation on SAOS-2 cells, from a human osteosarcoma cell line using a 3D titanium scaffold. 3D Ti6Al4V scaffolds were provided by Lima Lto (Udine, Italy). The human osteosarcoma cell line SAOS-2 was seeded on 3D Ti disks and cultured with or without (control condition) the electromagnetic stimulation (magnetic field,
linear finite element method (FEM). First, the ankle model (tibia–fibia–calcaneus–talus–trabecular–cortical bone and soft tissue) was developed with computer tomography, then the nonlinear FEM was applied to analyze the stresses and strains occurred in ligaments and bones for four different positions (10◦ Dorsiflexion, 0◦ Neutralposition, 10◦ Plantarflexion, 20◦ Plantarflexion). The ligaments were attached to the relevant points according to the anatomical structure of foot. The soft tissue was defined as hyper elastic while other tissues were idealized as homogeneous, isotropic and linearly elastic. Anterior displacement of foot from high to down in terms of different positions was found to be as 0◦ N > 20◦ PF > 10◦ DF > 10◦ PF respectively by applying 80 N force on calcaneus in horizontal direction. The maximum displacements were found on calcaneus where the loads were applied and the minumum displacements were found on fibula and tibia where the supports were located. The finite element method can be used as a powerful tool in the simulation of living structures. Using of noninvasive methods such as FEM can help to determine behaviour of ankle and ligaments before the treatment (ADT) applied to the patients. doi:10.1016/j.copbio.2011.05.502
R3 Plasmid DNA precipitation in the presence of cationic surfactant/salt: effect of the surface charge on precipitation Nese Ataci 1 , Ayfer Sarac 2 , Inci Arisan 1 1
Department of Biochemistry, Yildiz Technical University, Istanbul, Turkey 2 Department of Physical Chemistry, Yildiz Technical University, Istanbul, Turkey E-mail address: [email protected] (N. Ataci) DNA is a biological material, a kind of anionic polymer with double helical structure. Surfactants are an active field of research in colloidal science; especially consist of ionic or non-ionic head group and long alkyl chains. Many investigations have been presented on surfactant/DNA complex formation, precipitation and microstructure by a variety of methods. Knowledge of the interaction between DNA with cationic surfactants has many applications; the development of methods of DNA extraction and purification and lately, the potential use of these systems as vehicles for gene delivery and gene transfection. In this study, we investigated the effect of N,N ,N -polyethylene (10)-N tallow–1,3-di amine propane (PEG–10 tallow)/salt on plasmid DNA (pDNA) precipitation in critical micelle concentration (CMC). In the first step, CMC of PEG-10 tallow was determined by surface tension, and conductometry methods, and it was obtained closely agree with those determined both methods. The salt effect on precipitation profiles of PEG-10 tallow/pDNA formation was examined with agarose gel electrophoresis. Surface charges of PEG-10 tallow surfactant in different concentrations and surfactant/salt solutions were determined by zeta potential measurement. Results showed that, increased ionic strength changed parallel with DNA precipitation profiles. doi:10.1016/j.copbio.2011.05.503
S151
R4 Relative gene expression analysis shows increased mRNA levels of HoxB7 and HoxA10 and decreased mRNA levels of Pbx1 in the bone marrow derived mesenchymal progenitors from Fanconi anemia patients Aysen Gunel Ozcan 1 , Ilgin Cagnan 1 , Baris Kuskonmaz 2 , Duygu Uckan Cetinkaya 1 1
Hacettepe University, Institute of Pediatry & Center for Stem Cell Research and Development, Ankara, Turkey 2 Hacettepe University, Pediatric Hematology Bone Marrow Transplantation Unit, Ankara, Turkey E-mail address: [email protected] (A.G. Ozcan) Of having important roles on anterio-posterior body axis determination, embryonic body formation, hematopoeisis, eye development and determination of the cell type identity or cellular proliferation and differentiation during neuronal specification, Hox and the three-amino-acid loop extension family (TALE) proteins are the center of various biomedical research. Several research data shows that Hox gene family has great impact on regulating the genes and establishment of the balance between cellular proliferation and differentiation. Our research group aims to understand the Hox/TALE network and its regulators in Fanconi anemia, a cancer predisposition disorder characterized by progressive bone marrow failure. In this study, bone marrow derived mesenchymal stromal cells from three Fanconi anemia patients were expanded and characterized for their adipogenic and osteogenic differentiation. Spontaneous adipogenic differentiation was observed in one of the patients’ samples. Mesenchymal stromal cells of two patients did not differentiate to osteogenic lineage in osteoblast induction medium whereas donor cells did. Relative expression of Hox and Pbx1 (from TALE family) genes in mesenchymal stromal cells of patients and donors was determined by real-time RT-PCR. Analysis, performed by 2-((Ct method, showed that HoxB7 and HoxA10 mRNA levels in the patients’ cells increased whereas Pbx1 mRNA levels decreased. doi:10.1016/j.copbio.2011.05.513
R5 Strategies combining cells and scaffolds for bone tissue engineering Enrica Saino 1 , Nora Imbriani 4 , Livia Visai 5
Bloise 2 , Lorenzo
Fassina 3 , Marcello
1
Center for Tissue Engineering (C.I.T), Pavia, Italy Dep. Biochemistry, University of Pavia, Italy 3 Dep. Computer and Systems Science, University of Pavia, Italy 4 Salvatore Maugeri Foundation IRCCS, Pavia, Italy 5 International Centre for Studies and Research in Biomedicine (ICB), Luxembourg 2
E-mail address: [email protected] (E. Saino) Engineering bone uses porous scaffolds, osteoblasts or cells that can become osteoblasts, and factors that promote cell attachment, differentiation, and mineralized bone formation. In this study we investigated the effects of the electromagnetic stimulation on SAOS-2 cells, from a human osteosarcoma cell line using a 3D titanium scaffold. 3D Ti6Al4V scaffolds were provided by Lima Lto (Udine, Italy). The human osteosarcoma cell line SAOS-2 was seeded on 3D Ti disks and cultured with or without (control condition) the electromagnetic stimulation (magnetic field,