scholarly journals Influence of Culture Period on Osteoblast Differentiation of Tissue-Engineered Bone Constructed by Apatite-Fiber Scaffolds Using Radial-Flow Bioreactor

2021 ◽  
Vol 22 (23) ◽  
pp. 13080
Author(s):  
Kitaru Suzuki ◽  
Jun Fukasawa ◽  
Maiko Miura ◽  
Poon Nian Lim ◽  
Michiyo Honda ◽  
...  
Keyword(s):  
Osteoblast Differentiation ◽  
Radial Flow ◽  
Bone Marrow Cells ◽  
Three Dimensional ◽  
3D Cell Culture ◽  
Bone Diseases ◽  
Culture Period ◽  
Alizarin Red ◽  
Fiber Scaffolds ◽  
Radial Flow Bioreactor

With the limitation of autografts, the development of alternative treatments for bone diseases to alleviate autograft-related complications is highly demanded. In this study, a tissue-engineered bone was formed by culturing rat bone marrow cells (RBMCs) onto porous apatite-fiber scaffolds (AFSs) with three-dimensional (3D) interconnected pores using a radial-flow bioreactor (RFB). Using the optimized flow rate, the effect of different culturing periods on the development of tissue-engineered bone was investigated. The 3D cell culture using RFB was performed for 0, 1 or 2 weeks in a standard medium followed by 0, 1 or 2 weeks in a differentiation medium. Osteoblast differentiation in the tissue-engineered bone was examined by alkaline phosphatase (ALP) and osteocalcin (OC) assays. Furthermore, the tissue-engineered bone was histologically examined by hematoxylin and eosin and alizarin red S stains. We found that the ALP activity and OC content of calcified cells tended to increase with the culture period, and the differentiation of tissue-engineered bone could be controlled by varying the culture period. In addition, the employment of RFB and AFSs provided a favorable 3D environment for cell growth and differentiation. Overall, these results provide valuable insights into the design of tissue-engineered bone for clinical applications.

Reconstruction of Liver Organoid Using an Apatite-Fiber Scaffold, a Radial-Flow Bioreactor and FCL-4 Cells of Hepatocyte Model

2007 ◽  
Vol 361-363 ◽  
pp. 1165-1168 ◽  
Author(s):  
Mamoru Aizawa ◽  
A. Hiramoto ◽  
H. Maehashi ◽  
Tomokazu Matsuura
Keyword(s):  
Tissue Engineering ◽  
Single Crystal ◽  
Bone Tissue Engineering ◽  
Radial Flow ◽  
Three Dimensional ◽  
3D Culture ◽  
High Porosity ◽  
Fiber Scaffolds ◽  
Radial Flow Bioreactor ◽  
Culture Of Hepatocytes

We have previously developed apatite-fiber scaffolds (AFSs) for bone tissue engineering using single-crystal apatite fibers and carbon beads. In the present investigation, we examined the possibility of reconstruction of a liver organoid via three-dimensional (3D) culture of hepatocytes using the AFSs and the radial-flow bioreactor (RFB), aiming to apply the scaffold as a matrix for regeneration of a real organ. FLC-4 cells were used as a model of hepatocyte. The cells were well-viable in the RFB settled with AFSs over a period for 28 d, compared with the cases of cellulose beads and apatite beads with high porosity of 85%. We conclude that the present AFS may be a promising scaffold for tissue engineering of liver.

scholarly journals Production and release of infectious hepatitis C virus from human liver cell cultures in the three-dimensional radial-flow bioreactor

Virology ◽  
2003 ◽  
Vol 314 (1) ◽  
pp. 16-25 ◽  
Author(s):  
Hideki Aizaki ◽  
Seishi Nagamori ◽  
Mami Matsuda ◽  
Hayato Kawakami ◽  
Osamu Hashimoto ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Cell Cultures ◽  
Liver Cell ◽  
Human Liver ◽  
Radial Flow ◽  
Three Dimensional ◽  
Infectious Hepatitis ◽  
Human Liver Cell ◽  
Radial Flow Bioreactor

Osteogenic Effects of Triterpene Saponin Soyasapogenol B on Differentiation, Mineralization, Autophagy, and Necroptosis

2021 ◽  
Author(s):  
Kyung-Ran Park ◽  
Joon Yeop Lee ◽  
Soo Hyun Kim ◽  
Il Keun Kwon ◽  
Hyung-Mun Yun
Keyword(s):  
Cell Proliferation ◽  
Osteoblast Differentiation ◽  
Biological Effects ◽  
Bone Diseases ◽  
Adhesion Assay ◽  
Pharmacological Properties ◽  
Alizarin Red ◽  
Triterpenoid Saponins ◽  
Triterpene Saponin ◽  
Osteogenic Activity

Abstract Background: Triterpenoid saponins are a diverse group of natural compounds in plants. A triterpene saponin, Soyasapogenol B (SoyB), from Arachis hypogaea (peanut) has various pharmacological properties. This study aimed to elucidate pharmacological properties and mechanisms of SoyB on bone-forming cells. Methods: Cell viability adhesion, and migration were analyzed using MTT assay, cell adhesion assay, and Boyden chamber assay. Osteogenic activity and osteogenicity were analyzed using alkaline phosphatase (ALP) staining and activity, and Alizarin Red S (ARS) staining. Cell signaling, protein expression, and autophagy were analyzed using Western blot analysis, immunofluorescence assay, and DAPGreen autophagy detection assay. Results and Conclusion: In the present study, SoyB (> 99.99% purity), triterpene saponin, was isolated from the fruit of A. hypogaea. At concentrations ranging from 1 to 20 mM, SoyB showed no cell proliferation effects, whereas 30 - 100 mM SoyB increased cell proliferation in MC3T3-E1 cells. Next, osteoblast differentiation was analyzed and found that SoyB enhanced ALP staining and activity and bone mineralization as evidence for early and late osteoblast differentiation. SoyB also induced RUNX2 expression in nucleus with the increased phosphorylation of Smad1/5/8 and JNK2 during osteoblast differentiation. In addition, SoyB-mediated osteoblast differentiation was not associated with autophagy and necroptosis. Furthermore, SoyB increased cell migration and adhesion with the upregulation of MMP13 levels during osteoblast differentiation. The findings of this study provide new evidence that SoyB possesses biological effects on osteogenic activity and osteogenicity in bone-forming cells, and suggest a potentially beneficial role for peanuts foods and drugs containing SoyB in the treatment and prevention of bone diseases.

Gene expression property of high-density three-dimensional tissue of HepG2 cells formed in radial-flow bioreactor

2006 ◽  
Vol 101 (3) ◽  
pp. 243-250 ◽  
Author(s):  
Tomokatsu Hongo ◽  
Mariko Kajikawa ◽  
Seiichi Ishida ◽  
Shogo Ozawa ◽  
Yasuo Ohno ◽  
...  
Keyword(s):  
Gene Expression ◽  
Hepg2 Cells ◽  
Radial Flow ◽  
Three Dimensional ◽  
High Density ◽  
Radial Flow Bioreactor

scholarly journals Effects of Anhydroicaritin and 2″-Hydroxy-3″-en-Anhydroicaritin on the Proliferation and Differentiation of MC3T3–E1 Osteoblasts

2012 ◽  
Vol 7 (11) ◽  
pp. 1934578X1200701
Author(s):  
Yanping Li ◽  
Xuehui Zhang ◽  
Hui Peng ◽  
Rongtao Li ◽  
Xuliang Deng
Keyword(s):  
Regenerative Medicine ◽  
Immunosorbent Assay ◽  
Histochemical Staining ◽  
Bone Diseases ◽  
Culture Period ◽  
Enzyme Linked Immunosorbent Assay ◽  
Traditional Chinese Medicines ◽  
Alizarin Red ◽  
Chinese Medicines ◽  
Proliferation And Differentiation

Epimedium brevicornu Maxim., one of the most frequently used traditional Chinese medicines for thousands of years, is prescribed as having “bone strengthening” function and the ability to cure bone diseases. The present study evaluated the osteogenic effects of anhydroicaritin (1) and 2″-hydroxy-3″-en-anhydroicaritin (2) isolated from E. brevicornu by activity-guided assay. Treatment with1 and 2 improved the proliferation of murine osteoblastic MC3T3-E1 cells at doses of 10−7-10−5 mol/L and 10−7-10−6 mol/L, respectively, in the 72-hour culture period. Enzyme linked immunosorbent assay and histochemical staining demonstrated that both of these two prenyl-flavonoids significantly promoted the differentiation of MC3T3-E1 cells by enhancing the level of ALP activity in the cells. Alizarin Red staining and mineralized nodule quantification showed that 1 and 2 had the potential of stimulating the formation of mineralization nodules and further speeding up the formation of bone, indicating that both compounds might be potential candidates for bone regenerative medicine.

Three-dimensional high-density culture of HepG2 cells in a 5-ml radial-flow bioreactor for construction of artificial liver

2005 ◽  
Vol 99 (3) ◽  
pp. 237-244 ◽  
Author(s):  
Tomokatsu Hongo ◽  
Mariko Kajikawa ◽  
Seiichi Ishida ◽  
Shogo Ozawa ◽  
Yasuo Ohno ◽  
...  
Keyword(s):  
Hepg2 Cells ◽  
Radial Flow ◽  
Three Dimensional ◽  
High Density ◽  
Artificial Liver ◽  
High Density Culture ◽  
Radial Flow Bioreactor

scholarly journals TMARg, a Novel Anthraquinone Isolated from Rubia cordifolia Nakai, Increases Osteogenesis and Mineralization through BMP2 and β-Catenin Signaling

2020 ◽  
Vol 21 (15) ◽  
pp. 5332
Author(s):  
Kyung-Ran Park ◽  
Joon Yeop Lee ◽  
Bo-Mi Kim ◽  
Sang Wook Kang ◽  
Hyung-Mun Yun
Keyword(s):  
Cell Migration ◽  
Signaling Pathways ◽  
Intracellular Signaling ◽  
Osteoblast Differentiation ◽  
Biological Effects ◽  
Bone Diseases ◽  
C2c12 Cells ◽  
Alizarin Red ◽  
Rubia Cordifolia ◽  
Mitogen Activated Protein

Background: Plant extracts have long been regarded as useful medicines in the treatment of human diseases. Rubia cordifolia Nakai has been used as a traditional medicine, as it has pharmacological properties such as antioxidant and anti-inflammatory activity. However, the biological functions of TMARg, isolated from the roots of R. cordifolia, in osteoblast differentiation remain unknown. This study was performed to investigate the pharmacological effects and intracellular signaling of TMARg in the osteoblast differentiation of pre-osteoblast MC3T3-E1 cells and mesenchymal precursor C2C12 cells. Methods: Cell viability was evaluated using an MTT assay. Early and late osteoblast differentiation was examined by analyzing the activity of alkaline phosphatase (ALP), and by staining it with Alizarin red S (ARS). Cell migration was determined by using migration assays. Western blot analysis and immunocytochemical analysis were used to examine the intracellular signaling pathways and differentiation proteins. Results: In the present study, TMARg showed no cytotoxicity and increased the osteoblast differentiation in pre-osteoblasts, as assessed from the alkaline phosphate (ALP) staining and activity and ARS staining. TMARg also induced BMP2 expression and increased the p-smad1/5/8-RUNX2 and β-catenin pathways in both MC3T3-E1 and C2C12 cells. Furthermore, TMARg activated mitogen-activated protein kinases (MAPKs) and increased the cell migration rate. In addition, the TMARg-mediated osteoblast differentiation was suppressed by BMP and Wnt inhibitors with the downregulation of BMP2 expression. Conclusion: These findings demonstrate that TMARg exerts pharmacological and biological effects on osteoblast differentiation through the activation of BMP2 and β-catenin signaling pathways, and suggest that TMARg might be a potential phytomedicine for the treatment of bone diseases.

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