scholarly journals Studies on a Novel Bioreactor Design for Chondrocyte Culture

2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
Harshad Patil ◽  
Ishan Saurav Chandel ◽  
Amit K. Rastogi ◽  
Pradeep Srivastava
Keyword(s):  
Hydrodynamic Pressure ◽  
Airlift Reactor ◽  
Regular Growth ◽  
Basic Unit ◽  
Mixing Performance ◽  
Chondrocyte Culture ◽  
Cartilage Cells ◽  
Bioreactor System

A bioreactor system plays an important role in tissue engineering and enables reproduction and controlled changes in the environmental factor. The bioreactor provides technical means to perform controlled processes in safe and reduced reproducible generation of time. Cartilage cells were grown in vitro by mimicking the in vivo condition. The basic unit of cartilage, that is, chondrocyte, requires sufficient shear, strain, and hydrodynamic pressure for regular growth as it is nonvascular tissue. An attempt has been made to design a novel airlift reactor for chondrocyte culture, and the reactor has been evaluated for its performance. The design includes internal loop wavy riser airlift reactor for chondrocyte culture with 5% CO2 sparging which gives a good yield of chondrocyte after 28 days. The wavy riser provides more surfaces for collision of fluid flow so to create the turbulence. Also, the horizontal semicircular baffles create an angle of 180° which helps in high shear rate. The optimized L/D ratio of the designed airlift reactor (for chondrocyte culture) is 5.67, and it also exhibits good mixing performance.

In Vitro Chondrocyte Culture Using Hydrophilized PLLA Scaffold in Bioreactor System

2007 ◽  
Vol 342-343 ◽  
pp. 253-256 ◽  
Author(s):  
Hee Seok Yang ◽  
Kwi Deok Park ◽  
Byoung Soo Kim ◽  
Jae Jin Kim ◽  
Kwang Duk Ahn ◽  
...  
Keyword(s):  
3D Culture ◽  
Hydrodynamic Pressure ◽  
Chondrocyte Proliferation ◽  
Rabbit Knee ◽  
Plla Film ◽  
Chondrocyte Culture ◽  
Fetal Bovine ◽  
Bioreactor System

Nonporous PLLA film and porous PLLA scaffolds were prepared and then grafted with acrylic acid (AA) using in situ direct plasma treatment to obtain PLLA-g-PAA. Chondrocytes isolated from rabbit knee articular cartilages were cultivated in Dulbecco’s modified eagle medium- F12 (DMEM-F12) containing 10% fetal bovine serum (FBS) and 1% antibiotics and passaged twice before cell seeding. Once seeded on either PLLA films or scaffolds, they were placed in a bioreactor system and an intermittent hydrodynamic pressure (IHP) was applied in 3 bars, while turned on for 2 min and off for 28 min during 15-day culture. AA grafting to PLLA surface was confirmed from various surface analyses. From WST-1 assay, chondrocyte proliferation was significantly improved with dynamic IHP for PLLA and PLLA-g-PAA scaffolds as compared to static culture. This study indicates that IHP may have significant influence on chondrocytes behavior in 3D culture environment.

Dexamethasone induces apoptosis in proliferative canine tendon cells and chondrocytes

2008 ◽  
Vol 21 (04) ◽  
pp. 337-342 ◽  
Author(s):  
M. A. Hossain ◽  
J. Park ◽  
S. H. Choi ◽  
G. Kim
Keyword(s):  
Cell Proliferation ◽  
Cartilage Degeneration ◽  
Dependent Manner ◽  
Tendon Cells ◽  
Cartilage Cells ◽  
In Vitro Effects ◽  
Nuclear Apoptosis ◽  
And Cartilage

SummaryDexamethasone (Dexa) has been commonly used in humans and domestic animals, particularly in the treatment of tendon injuries and cartilage degeneration. However, it is often associated with tendon rupture and impaired tendon and cartilage healing. In the present study, we investigated Dexa’s in vitro effects on the growth of cell proliferation and the induction of apoptosis in canine Achilles tendon cells and chondrocytes. Cell proliferation after treatment with Dexa for two to six days was quantified by a 2,3-bis{2-methoxy- 4-nitro-5-sulfophenyl}-2H-tetrazolium-5-carboxyanilide inner salt assay (XTT). The results showed that Dexa could inhibit the proliferation of tendon cells and chondrocytes at increasing concentrations (0.1–50 μg/ml) compared with untreated cells. Cell apoptosis was induced by Dexa, as evidenced by the typical nuclear apoptosis using Hoechst 33258 staining. Dexa increased the apoptosis of canine tendon cells and chondrocytes in a time-dependent manner. In canine tendon cells and chondrocytes that were treated with 25 and 50 μg/ml concentration of Dexa, the number of condensed apoptotic nuclei was significantly increased. In addition, culturing with Dexa and the glucocorticoid receptor blocker, mifepristone, significantly arrested apoptosis of tendon cells and chondrocytes. Based on our in vitro data, we hypothesized that in vivo treatment with glucocorticoids may diminish the proliferation of tendon and cartilage cells by increasing apoptosis and suppressing the proliferation. Our findings suggest that Dexa could be used with caution in dogs with articular or tendon problems.

scholarly journals Validation of a novel perfusion bioreactor system in cancer research

2020 ◽  
Vol 74 (3) ◽  
pp. 187-196
Author(s):  
Jasmina Stojkovska ◽  
Jovana Zvicer ◽  
Milena Milivojevic ◽  
Isidora Petrovic ◽  
Milena Stevanovic ◽  
...  
Keyword(s):  
Cancer Research ◽  
Drug Screening ◽  
Cancer Drug ◽  
Perfusion Bioreactor ◽  
Animal Testing ◽  
Anti Cancer ◽  
Medium Flow ◽  
Bioreactor System

Development of drugs is a complex, time- and cost-consuming process due to the lack of standardized and reliable characterization techniques and models. Traditionally, drug screening is based on in vitro analysis using two-dimensional (2D) cell cultures followed by in vivo animal testing. Unfortunately, application of the obtained results to humans in about 90 % of cases fails. Therefore, it is important to develop and improve cell-based systems that can mimic the in vivo-like conditions to provide more reliable results. In this paper, we present development and validation of a novel, user-friendly perfusion bioreactor system for single use aimed for cancer research, drug screening, anti-cancer drug response studies, biomaterial characterization, and tissue engineering. Simple design of the perfusion bioreactor provides direct medium flow at physiological velocities (100?250 ?m s-1) through samples of different sizes and shapes. Biocompatibility of the bioreactor was confirmed in short term cultivation studies of cervical carcinoma SiHa cells immobilized in alginate microfibers under continuous medium flow. The results have shown preserved cell viability indicating that the perfusion bioreactor in conjunction with alginate hydrogels as cell carriers could be potentially used as a tool for controlled anti-cancer drug screening in a 3D environment.

scholarly journals A new principle of oligomerization of plant DEG7 protease based on interactions of degenerated protease domains

2011 ◽  
Vol 435 (1) ◽  
pp. 167-174 ◽  
Author(s):  
Holger Schuhmann ◽  
Ulrike Mogg ◽  
Iwona Adamska
Keyword(s):  
Basic Unit ◽  
Pdz Domains ◽  
Protease Domain ◽  
Yeast Two Hybrid ◽  
Protein Protein Interaction ◽  
Plant Protease ◽  
Active Protease ◽  
Two Hybrid

Deg/HtrA proteases are a large group of ATP-independent serine endoproteases found in almost every organism. Their usual domain arrangement comprises a trypsin-type protease domain and one or more PDZ domains. All Deg/HtrA proteases form homo-oligomers with trimers as the basic unit, where the active protease domain mediates the interaction between individual monomers. Among the members of the Deg/HtrA protease family, the plant protease DEG7 is unique since it contains two protease domains (one active and one degenerated) and four PDZ domains. In the present study, we investigated the oligomerization behaviour of this unusual protease using yeast two-hybrid analysis in vivo and with recombinant protein in vitro. We show that DEG7 forms trimeric complexes, but in contrast with other known Deg/HtrA proteases, it shows a new principle of oligomerization, where trimerization is based on the interactions between degenerated protease domains. We propose that, during evolution, a duplicated active protease domain degenerated and specialized in protein–protein interaction and complex formation.

scholarly journals Finite element study of stem cells under fluid flow for mechanoregulation toward osteochondral cells

2021 ◽  
Vol 32 (7) ◽  
Author(s):  
Mehdi Moradkhani ◽  
Bahman Vahidi ◽  
Bahram Ahmadian
Keyword(s):  
Stem Cells ◽  
Shear Stress ◽  
Fluid Flow ◽  
Computational Simulation ◽  
Hydrodynamic Pressure ◽  
Mechanical Stimuli ◽  
Pore Architecture ◽  
Proliferation And Differentiation

AbstractInvestigating the effects of mechanical stimuli on stem cells under in vitro and in vivo conditions is a very important issue to reach better control on cellular responses like growth, proliferation, and differentiation. In this regard, studying the effects of scaffold geometry, steady, and transient fluid flow, as well as influence of different locations of the cells lodged on the scaffold on effective mechanical stimulations of the stem cells are of the main goals of this study. For this purpose, collagen-based scaffolds and implicit surfaces of the pore architecture was used. In this study, computational fluid dynamics and fluid-structure interaction method was used for the computational simulation. The results showed that the scaffold microstructure and the pore architecture had an essential effect on accessibility of the fluid to different portions of the scaffold. This leads to the optimization of shear stress and hydrodynamic pressure in different surfaces of the scaffold for better transportation of oxygen and growth factors as well as for optimized mechanoregulative responses of cell–scaffold interactions. Furthermore, the results indicated that the HP scaffold provides more optimizer surfaces to culture stem cells rather than Gyroid and IWP scaffolds. The results of exerting oscillatory fluid flow into the HP scaffold showed that the whole surface of the HP scaffold expose to the shear stress between 0.1 and 40 mPa and hydrodynamics factors on the scaffold was uniform. The results of this study could be used as an aid for experimentalists to choose optimist fluid flow conditions and suitable situation for cell culture.

scholarly journals 225 RECOMBINANT HUMAN ERYTHROPOIETIN PRODUCED IN THE MILK OF TRANSGENIC MICE: FUNCTIONAL CHARACTERIZATION AND PHARMACEUTICAL APPROACH

2005 ◽  
Vol 17 (2) ◽  
pp. 263
Author(s):  
D.N. Kwon ◽  
J.Y. Park ◽  
S.Y. Lee ◽  
S.J. Kang ◽  
J.H. Kim
Keyword(s):  
Mammary Gland ◽  
Transgenic Mouse ◽  
Cho Cells ◽  
Recombinant Human Erythropoietin ◽  
Mouse Mammary Gland ◽  
Human Erythropoietin ◽  
Mouse Milk ◽  
Bioreactor System

The proper post-transcriptional modification of recombinant human erythropoietin (rhEPO) is critical to retain its biological functions, either in vivo or in vitro. The major glycosyltransferases for the determinant of glycosylation patterns of rhEPO are N-acetylglycosaminyltransferase (GnT) and α-1-3/4 fucosyltransferase (Fut). GnT-III expression (388 ± 19.09) in the mouse mammary gland has been shown to be dramatically different from that in CHO cells, although FuT-VIII expression in CHO cells (1970 ± 255.9) is comparable to mouse mammary gland (272 ± 14.8), suggesting that the mammary gland may proceed with the proper glycosylation of rhEPO as shown in CHO cells. To identify this hypothesis and establish the rhEPO bioreactor system for mass production of protein in transgenic animals, we have generated two transgenic mouse lines that express rhEPO in milk. Both lines of transgenic mouse express only rhEPO in the lactating mammary gland, and the protein yield of rhEPO in lactating milk is comparable to that in CHO cells. After determining the protein expression in lactating milk, using three different methods – enzymatic release of oligosaccharide analysis, two-dimensional electrophoresis, and 2-aminobenzamide-labeled analysis – we report that the rhEPO produced by the animal bioreactor system has the proper glycosylation patterns as shown in CHO cell-derived Epoietin α, and has more tetra-acidic oligosaccharide structures than Epoietin α, which is the widely used rhEPO for therapeutic purposes. The in vitro biological property of transgenic mouse milk-derived rhEPO has been tested by measuring luciferase activity in MCF-7 cells, indicating that rhEPO from mammary gland up-regulates the EPO-receptor-mediated STAT5 gene expression in a dose-dependent manner the same as Epoietin α does. In addition, in vivo biological activity demonstrated that direct injection of rhEPO into a mouse vein increases blood components such as RBC and HCT. In light of these findings, we suggest that high levels of tetra-acidic structures observed in transgenic mouse milk-derived rhEPO may be related to the high level of expression of glycosiltransferases (GnT-III and FuT-VIII) in mammary gland; thus the bioreactor system using the mammary gland of a transgenic animal could be a good candidate for production of rhEPO for pharmaceutical purposes. This work was supported in part by a grant program from RDA(Biogreen21) and Cho-A, Republic of Korea. D.N. Kwon is the recipient of a scholarship from the BK21 program, granted by the Ministry of Education, Korea.

scholarly journals Development and implementation of the GIT-modelling bioreactor system: the way to reducing a carbon footprint

2021 ◽  
Vol 279 ◽  
pp. 01030
Author(s):  
Donskoy Danila ◽  
Katin Oleg ◽  
Alekseenko Ludmila
Keyword(s):  
Gastrointestinal Tract ◽  
Carbon Footprint ◽  
Nutrient Medium ◽  
Broiler Chicken ◽  
Experimental Results ◽  
In Vitro Experiments ◽  
Control Computer ◽  
Bioreactor System

The purpose of this article is to analyze and compare the carbon footprint value between «in vivo» and «in vitro» microbiological experiments in chicken GIT. The SHIME-style bioreactor for modeling the processes occurring in the gastrointestinal tract of chickens is developed. For «in vitro» estimation – use analytic and experimental results, carrying out on developed chicken GIT model. For « in vivo» estimation, use the carbon footprint of two mounts old broiler chicken. Assessments showed that «in vitro» carbon footprint constitutes about 15% of «in vivo» one. The most significant contributors to «in vitro» carbon footprint are nutrient medium (80%), then control computer (10%), then heather (5%). Recommendations for further carbon footprint reduction for «in vitro» experiments are formulated.

Anti-idiotypic antibody as an oestrogen mimetic: removal of Fc fragment converts agonist to antagonist

1995 ◽  
Vol 145 (3) ◽  
pp. 409-416 ◽  
Author(s):  
D Sömjen ◽  
Y Amir-Zaltsman ◽  
B Gayer ◽  
G Mor ◽  
N Jaccard ◽  
...  
Keyword(s):  
Physiological Responses ◽  
Female Rats ◽  
Dependent Manner ◽  
Immature Female ◽  
Antibody Molecule ◽  
Cartilage Cells ◽  
Dose Dependent ◽  
Idiotypic Antibody

Abstract Previous studies indicated that the anti-idiotypic antibody (clone 1D5) caused an increase in uterine creatine kinase (CK) activity when administered in vivo to immature female rats, indicating that the antibody has oestrogenic-like activity. It was, therefore, of interest to investigate the structural requirements of clone 1D5 to act as an oestrogen mimetic in an in vitro model system. In the present study, the effect of clone 1D5 and its proteolytic fragments, F(ab′)2, Fab′ and Fc on CK activity was examined in cultured skeletal cells having functional oestrogen receptor (ER). Incubation of female-derived calvaria cells or epiphyseal cartilage cells with clone 1D5 (8·33 nm) or oestradiol (E2) (30 nm) for 24 h caused a significant increase in CK activity, indicating that clone 1D5 acted as an agonist. On the other hand, incubation of male-derived calvaria cells devoid of a functional ER with clone 1D5 or E2 did not have any effect on CK activity. Incubation of female-derived calvaria cells with clone 1D5 and E2 did not result in any further increase in CK activity, whereas dihydrotestosterone (DHT) did not alter the response to clone 1D5. The CK response to clone 1D5, in female-derived calvaria cells was time- and dose-dependent and could be inhibited in a dose-dependent manner by the oestrogen antagonist tamoxifen. In contrast, the proteolytic fragments of clone 1D5, the F(ab′)2 dimer (12 nm) and the Fab′ monomer (24 nm), and the Fc fragment (28 nm) did not have E2-like activity in these cells. However, while the Fab′ monomer or the Fc fragment, as well as clone 1D5, did not affect the response of the female-derived calvaria cells to E2, the F(ab′)2 dimer acted like an antagonist and completely inhibited the stimulatory effect of E2 or 1D5, but was unable to block the stimulatory effect of DHT on CK in male-derived calvaria cells. Collectively, these results imply that a bivalent antibody is necessary for the observed physiological responses, and that the anti-idiotypic antibody can be converted from an agonist to an antagonist by removal of the Fc portion of the antibody molecule. Furthermore, the anti-idiotypic antibody has an oestrogenic-like effect inhibited by tamoxifen only in skeletal cells capable of responding to E2. Journal of Endocrinology (1995) 145, 409–416

scholarly journals In vivo bioreactor using cellulose membrane benefit engineering cartilage by improving the chondrogenesis and modulating the immune response

2019 ◽  
Author(s):  
Xue Guang Li ◽  
In-Su Park ◽  
Byung Hyune Choi ◽  
Ung-Jin Kim ◽  
Byoung-Hyun Min
Keyword(s):  
Immune Response ◽  
Biochemical Analysis ◽  
Cartilage Defects ◽  
Cellulose Membrane ◽  
Rt Pcr ◽  
Bioreactor System ◽  
Regenerate Tissue ◽  
Engineered Cartilage

ABSTRACTTo regenerate tissue engineered cartilage as a source for the restoration of cartilage defects, we used a human fetal cartilage progenitor cell (hFCPC) pellet for improve the chondrogenesis and modulation the immune response with a In vivo (IV) bioreactor system, that was buried subcutaneously in the host and then implanted into a cartilage defect. In vivo bioreactor (IVB) was composed of silicone tube and cellulose nanopore-size membrane. FCPC pellets were first cultured in vitro for 3 days, and then cultured in vitro, subcutaneous and IV bioreactor for 3 weeks. First evaluated the IV bioreactor fluid appearance, component and liquidity, and then evaluate chondrogenesis and immunogenicity of the pellets using gross observation, cell viability, histology, biochemical analysis, RT-PCR, and Western Blot, finally evaluates the cartilage repair and synovial inflammation using histology. The fluid color and transparency of IV bioreactor were similar to synovial fluid (SF) and the component was also close to SF compared to the serum. IV bioreactor system not only promotes the synthesis of cartilage matrix and maintains cartilage phenotype, but also delays the occurrence of calcification compared with subcutaneous. A IV bioreactor, which has been predominantly adopted to study cell differentiation, was effective in preventing host immune rejection.

scholarly journals The biological reaction of arterial pressure and pathogenesis of programmed arterial hypertension

2011 ◽  
Vol 17 (6) ◽  
Author(s):  
V. N. Titov
Keyword(s):  
Body Weight ◽  
Arterial Hypertension ◽  
Biological Function ◽  
Hydrodynamic Pressure ◽  
Postnatal Period ◽  
Compensatory Hypertrophy ◽  
Basic Unit ◽  
Nephron Development ◽  
Cell Community

Phylogenetically, the following scheme is relevant in vivo: cell → cell community → structural unit of an organ → organ → organ system. A paracrine-regulated cell community is a structural and functional unit of an internal organ; the nephron composed of renal capillary, afferent (efferent) muscular arteriole, and paratubular loose connective tissue is the basic unit of the kidney. In primates the nephron is induced from three germ layers within the first 16 weeks of embryogenesis, which is followed by nephron development during prenatal period; neonephrogenesis does not occur in the postnatal period. Impaired biological function of exotrophy, protein deficiency in mother's diet, and glucocorticoid therapy reduce the number of nephrons, which is reflected by low body weight of the newborn. If a newborn weights less than 2,5 kg, the number of nephrons is so small that they cannot provide the biological function of endoecology in adults without compensatory hypertrophy, which leads to the hydrodynamic pressure elevation in the arterial bed over glomerular filter and increased glomerular filtration. However, hypertrophy is not optimal for compensation of the nephron function. Low body weight at birth and small number of nephrons are risk factors of programmed arterial hypertension in an adult organism.

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