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.

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.

scholarly journals Evaluation of Osteogenic Potentials of Titanium Dioxide Nanoparticles with Different Sizes and Shapes

2020 ◽  
Vol 2020 ◽  
pp. 1-13
Author(s):  
Yixing Ren ◽  
Xinxing Feng ◽  
Xinhua Lang ◽  
Jianbo Wang ◽  
Zhipo Du ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Physicochemical Properties ◽  
Titanium Dioxide Nanoparticles ◽  
Enzyme Linked Immunosorbent Assay ◽  
Alizarin Red ◽  
Tio2 Nps ◽  
Transmission Electron ◽  
Proliferation And Differentiation ◽  
Different Shapes ◽  
The Impact

TiO2 nanoparticles (NPs) have the potential to be used in the human body as an artificial implant because of their special physicochemical properties. However, information about the effects of TiO2 NPs on preosteoblast proliferation and osteogenic differentiation is not clear. In this work, we focus on the impact of TiO2 NPs with different shapes and sizes on the proliferation and differentiation of MC3T3-E1 cells. The morphology and physicochemical properties of TiO2 NPs are analyzed by scanning electron microscopy, transmission electron microscopy, Quadrasorb SI analyzer, dynamic light scattering, and zeta potential. The MTT results indicate that when the concentration of TiO2 NPs is less than 20 μg/mL, the proliferation of osteoblasts is preserved the most. The expression of alkaline phosphatase and osteocalcin is detected by BCA and enzyme-linked immunosorbent assay to analyze the differentiation of osteoblasts. The results indicate that both rutile and anatase TiO2 NPs have a significant inhibiting influence on the differentiation of osteoblasts. Alizarin Red staining is performed on cells to detect the mineralized nodules. The results show that TiO2 NPs can promote the mineralization of MC3T3-E1 cells. Then, we study the oxidative stress response of MC3T3-E1 cells by flow cytometry analysis, and all TiO2 NPs induce the excessive generation of reactive oxide species. On the other hand, our study also shows that the early apoptotic cells increase significantly. TiO2 NPs are swallowed by cells, and then the agglomerated particles enter mitochondria, causing the shape of mitochondria to change and vacuolation to appear. All these results show that TiO2 NPs have certain cytotoxicity to cells, but they also promote the mineralization and maturation of osteoblasts.

Plasminogen Interactions with Immobilized Fibrinogen

1989 ◽  
Vol 62 (04) ◽  
pp. 1078-1082 ◽  
Author(s):  
Burt Adelman ◽  
Patricia Ouynn
Keyword(s):  
Immunosorbent Assay ◽  
Aminocaproic Acid ◽  
Enzyme Linked Immunosorbent Assay ◽  
Binding Regions ◽  
Dose Dependent Fashion ◽  
Epsilon Aminocaproic Acid ◽  
Dose Dependent

SummaryThis report describes the binding of plasminogen to fibrinogen adsorbed onto polystyrene wells. Binding was determined by enzyme linked immunosorbent assay. Both glu- and lys-plasminogen bound to immobilized fibrinogen in a dose-dependent fashion. However, more lys- than glu-plasminogen bound when equal concentrations of either were added to immobilized fibrinogen. Plasminogen binding was inhibited by epsilon aminocaproic acid indicating that binding was mediated via lysine-binding regions of plasminogen. Soluble fibrinogen added in excess of immobilized fibrinogen did not compete for plasminogen binding but fibrinogen fragments produced by plasmin digestion of fibrinogen did. Treatment of immobilized fibrinogen with thrombin caused a small but significant (p <0.01) increase in plasminogen binding. These studies demonstrate that immobilized fibrinogen binds both glu- and lys-plasminogen and that binding is mediated via lysine-binding regions. These interactions may facilitate plasminogen binding to fibrinogen adsorbed on to surfaces and to cells such as platelets which bind fibrinogen.

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