scholarly journals The Study on Biocompatibility of Porous nHA/PLGA Composite Scaffolds for Tissue Engineering with Rabbit ChondrocytesIn Vitro

2013 ◽  
Vol 2013 ◽  
pp. 1-6 ◽  
Author(s):  
Lei Chen ◽  
Wei-Min Zhu ◽  
Zhi-Qiang Fei ◽  
Jie-Lin Chen ◽  
Jian-Yi Xiong ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Tissue Engineering ◽  
Flow Cytometry ◽  
Mtt Assay ◽  
Cartilage Tissue Engineering ◽  
Cartilage Tissue ◽  
Control Group ◽  
Dependent Manner ◽  
Plga Scaffold ◽  
Significant Difference

Objective. To examine the biocompatibility of a novel nanohydroxyapatite/poly[lactic-co-glycolic acid] (nHA/PLGA) composite and evaluate its feasibility as a scaffold for cartilage tissue engineering.Methods. Chondrocytes of fetal rabbit were cultured with nHA/PLGA scaffoldin vitroand the cell viability was assessed by MTT assay first. Cells adhering to nHA/PLGA scaffold were then observed by inverted microscope and scanning electron microscope (SEM). The cell cycle profile was analyzed by flow cytometry.Results. The viability of the chondrocytes on the scaffold was not affected by nHA/PLGA comparing with the control group as it was shown by MTT assay. Cells on the surface and in the pores of the scaffold increased in a time-dependent manner. Results obtained from flow cytometry showed that there was no significant difference in cell cycle profiles between the coculture group and control (P>0.05).Conclusion. The porous nHA/PLGA composite scaffold is a biocompatible and good kind of scaffold for cartilage tissue engineering.

Effects of 5-Bromotetrandrine and Daunorubicin on the Expression of Survivin In K562/AO2 Cells

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 4947-4947
Author(s):  
Bao-An Chen ◽  
Xiao-hui Cai ◽  
Jun Wang ◽  
Chong Gao ◽  
Jia-hua Ding ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Mtt Assay ◽  
Conflicts Of Interest ◽  
Acquired Resistance ◽  
Leukemic Cells ◽  
Control Group ◽  
Dependent Manner ◽  
Rt Pcr ◽  
Reverse Transcription Pcr ◽  
Typical Experiment

Abstract Abstract 4947 Objective: The aim of this study was to investigate the expression of survivin and the apoptosis induced by DNR and BrTet in the leukemic cells K562/A02. Methods: In a typical experiment, the K562/AO2 cells were treated with daunorubicin (DNR), 5-bromotetrandrine (BrTet), or DNR and BrTet for 48 hours, and the cells treated without any drugs were used as control group. Cell proliferation was analyzed by MTT assay. Cells apoptosis and the concentration of DNR within the cells were measured by Flow cytometry (FCM). The expressions of mRNA and protein of survivin were determined by semi-quantitative reverse transcription PCR (RT-PCR) and Western blot, respectively. Results: The results of MTT assay indicated that DNR and BrTet were both able to inhibit the proliferation of K562/AO2 cells in dose-dependent manner. The fresh evidence from flow cytometry showed that a higher apoptosis rate could be induced and a higer concentration of DNR could be detected in K562/AO2 cells by DNR and BrTet as compared with those by DNR or BrTet in the same concentrations(P<0.01). RT-PCR revealed that the expression of survivin mRNA, a higer expression in K562/AO2 cells with acquired resistance to adriamycin than that in parent K-562 cells, decreased in the DNR and BrTet group (P<0.05), but there was no obvious change in other groups(P>0.05). Western bolt demonstrated that the expression of survivin protein was much lower in the DNR and BrTet group(P<0.05). Conclusion: BrTet could increase the concentration of DNR and reverse the multidrug resistance(MDR) in the K562/AO2 cells. Survivin may play an important role in apoptosis induced by DNR. Survivin could be a target for the treatment of MDR in haematopoietic malignancies. Disclosures: No relevant conflicts of interest to declare.

Rapamycin Induced Autophagy and Suppression in Human Leukemia Cells Proliferation

2014 ◽  
Vol 926-930 ◽  
pp. 1112-1115
Author(s):  
Yi Ju Hou ◽  
Zhong Hai Yuan ◽  
Xiao Dong Liu ◽  
Ke Xin Sun ◽  
Chen Zhao ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
Cell Proliferation ◽  
Mtt Assay ◽  
Leukemia Cells ◽  
Control Group ◽  
Human Leukemia ◽  
Hl60 Cells ◽  
Human Leukemia Cells ◽  
Autophagic Activity

To investigate the effect of autophagy induced by rapamycin (Rap) on proliferation of Human Leukemia Cells (HL60). HL60 cells are treated with rapamycin to induce autophagy. Then, Western blot is performed to examine the expression of Beclin1 and LC3. MTT assay is used to evaluate cell proliferation. The cell cycle is analyzed by flow cytometry (FCM). After treatment with rapamycin for 24 hours, LC3 is dramatically increased at protein level and autophagic activity is significantly increased. And MTT assay indicated that cell proliferation is inhibited by rapamycin. Compared with control group, more cells are arrested at G0/ G1 phases. We conclude that rapamycin can induce autophagy and suppress proliferation in HL60 cells.

scholarly journals Coculture of allogenic DBM and BMSCs in the knee joint cavity of rabbits for cartilage tissue engineering

2017 ◽  
Vol 37 (6) ◽  
Author(s):  
Bin Xu ◽  
Rui Wang ◽  
Hao Wang ◽  
Hong-Gang Xu
Keyword(s):  
Tissue Engineering ◽  
Knee Joint ◽  
Cartilage Tissue Engineering ◽  
Cartilage Tissue ◽  
Control Group ◽  
Blank Control Group ◽  
Joint Cavity ◽  
A Value

The present study aims to assess coculture of allogenic decalcified bone matrix (DBM) and bone marrow mesenchymal stem cells (BMSCs) in the knee joint cavity of rabbits for cartilage tissue engineering. Rabbits were assigned to an in vitro group, an in vivo group, and a blank control group. At the 4th, 8th, and 12th week, samples from all groups were collected for hematoxylin–eosin (HE) staining and streptavidin–peroxidase (SP) method. The morphological analysis software was used to calculate the average absorbance value (A value). SP and flow cytometry demonstrated that BMSCs were induced into chondrocytes. DBM scaffold showed honeycomb-shaped porous and three-dimensional structure, while the surface pores are interlinked with the deep pores. At the 4th week, in the blank control group, DBM scaffold structure was clear, and cells analogous to chondrocytes were scattered in the interior of DBM scaffolds. At the 8th week, in the in vivo group, there were a large amount of cells, mainly mature chondrocytes, and the DBM scaffolds were partially absorbed. At the 12th week, in the in vitro group, the interior of scaffolds was filled up with chondrocytes with partial fibrosis, but arranged in disorder. In the in vivo group, the chondrocytes completely infiltrated into the interior of scaffolds and were arranged in certain stress direction. The in vivo group showed higher A value than the in vitro and blank control groups at each time point. Allogenic DBM combined BMSCs in the knee joint cavity of rabbits could provide better tissue-engineered cartilage than that cultivated in vitro.

Involvement of NF-κB activation in the apoptosis induced by extracellular adenosine in human hepatocellular carcinoma HepG2 cellsThis paper is one of a selection of papers published in this special issue entitled “Second International Symposium on Recent Advances in Basic, Clinical, and Social Medicine” and has undergone the Journal's usual peer review process.

2010 ◽  
Vol 88 (4) ◽  
pp. 705-714 ◽  
Author(s):  
Ling-Fei Wu ◽  
Guo-Ping Li ◽  
Jian-Dong Su ◽  
Ze-Jin Pu ◽  
Jia-Lin Feng ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Cycle ◽  
Flow Cytometry ◽  
Mtt Assay ◽  
Cell Apoptosis ◽  
Hepg2 Cells ◽  
Caspase 3 ◽  
Human Hepatocellular Carcinoma ◽  
Pyrrolidine Dithiocarbamate ◽  
Dependent Manner

Adenosine can exhibit cytotoxic activity in vivo and in vitro, though its mechanisms are still uncertain. In this study, we investigated the adenosine-mediated apoptotic signaling pathway and the role of NF-κB in human hepatocellular carcinoma HepG2 cells. HepG2 cells were treated with different concentrations of adenosine for 12–48 h, and the effect of adenosine on cell proliferation was evaluated by MTT assay. The cytotoxicity of adenosine alone or in combination with an NF-κB inhibitor, pyrrolidine dithiocarbamate (PDTC), was also evaluated by MTT assay and the mode of cell death was detected by Hoechst 33342 staining. Cell cycle progress was performed by flow cytometry with PI staining. The protein expressions of Bcl-2, p53, NF-κB subunit p65, and caspase-3 were assayed by Western blot. Caspase-3 activity was measured by spectrophotomteric assay. The results showed that adenosine significantly reduced the viability of HepG2 cells in a dose- and time-dependent manner, with IC 50 (24 and 48 h) of 2.52 and 1.89 mmol·L–1, respectively. The apoptotic index (percentage of sub-G1 phase) of HepG2 cells in adenosine treatment alone for 12 and 24 h or in combination with PDTC were 8.30%, 22.32% and 20.18%, 30.89%, respectively. All of them were higher than that in the control group (0.81%, p < 0.01). The characteristic changes of cell apoptosis (chromatin condensation and sub-G1 peak) were observed under fluorescent microscopy and flow cytometry. We also found that the apoptotic process triggered by adenosine was involved in G0–G1 cell-cycle arrest, enhanced the activity of caspase-3, upregulated p53 and NF-κB p65 expression, and downregulated Bcl-2 expression. Inhibition of NF-κB by PDTC decreased NF-κB p65 expression, enhanced cell apoptosis ratio, and increased caspase-3 activity. NF-κB may play an anti-apoptosis role in adenosine-induced HepG2 cytotoxicity.

scholarly journals Dose-Dependent Effect of Cordycepin on Viability, Proliferation, Cell Cycle, and Migration in Dental Pulp Stem Cells

2021 ◽  
Vol 11 (8) ◽  
pp. 718
Author(s):  
Nezar Boreak ◽  
Ahmed Alkahtani ◽  
Khalid Alzahrani ◽  
Amani Hassan Kenani ◽  
Wafa Hussain Faqehi ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Flow Cytometry ◽  
Growth Curve ◽  
Dental Pulp ◽  
Phase Cell ◽  
Ros Generation ◽  
Dependent Manner ◽  
Significant Difference ◽  
Dose Dependent

Objective: To examine the effect of Cordycepin on the viability, proliferation, and migratory properties of dental pulp-derived mesenchymal stem cells. Materials and methods: The pulp was derived from human premolar teeth extracted for orthodontic purposes after obtaining informed consent. The samples were transferred to the laboratory for processing. DPSCs were expanded and characterized using flow cytometry and differentiation to the bone, adipose, and cartilage cells was examined. MTT Assay was performed using various concentrations of Cordycepin. The growth curve was plotted for 13 days. Cell cycle analysis was performed by flow cytometry. Migratory ability was assessed by wound healing assay. ROS generation was detected by flow cytometry. Gene expression was quantified by RT-qPCR. Statistical analysis was performed. p < 0.05 was considered as significant and p < 0.01 was considered as highly significant (* p < 0.05, and ** p < 0.01). Results: DPSCs expressed characteristic MSC-specific markers and trilineage differentiation. Cordycepin at lower concentrations did not affect the viability of DPSCs. The growth curve of cells showed a dose-dependent increase in cell numbers till the maximum dose. DPSCs treated with 2.5 µM Cordycepin was found to have a reduced G1 phase cell percentage. DPSCs treated with 2.5 µM and 5 µM Cordycepin showed a significant decrease in G2 phase cells. No significant difference was observed for S phase cells. Cordycepin treatment affected the migratory ability in DPSCs in a concentration-dependent manner. Conclusion: Cordycepin can be used at therapeutic doses to maintain stem cells.

The Study of Effect and Mechanism by Vitamin K2 on Human MDS Cell Line MUTZ-1.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4357-4357
Author(s):  
Bao-An Chen ◽  
Xin Xu ◽  
Ze-Ye Shao ◽  
Jia-Hua Ding ◽  
Guo-Hua Xia ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Line ◽  
Caspase 3 ◽  
Cellular Proliferation ◽  
Vitamin K2 ◽  
Time Dependent ◽  
Control Group ◽  
Dependent Manner ◽  
Significant Difference

Abstract The myelodysplastic syndromes (MDS) are characterized by hemopoietic insufficiency associated with cytopenias leading to serious morbidity and the additional risk of leukemic transformation. Vitamin K2(VK2) is reported to induce apoptosis or differentiation of leukemic cell lines in vitro. For investigating the effects and mechanism of VK2 on human MDS cell line MUTZ-1 in vitro,we observed the changes of morphologic features of MUTZ-1 cells by exposing to VK2.The transmission electron microscope was used to observe the apoptosis of MUTZ-1 cells. Cellular proliferation was determined by the MTT assay. The flow cytometry was used to analysis apoptosis rate and the change of cell cycle. The expression of apoposis-related genes bcl-2, survivin and bax were detected by reverse transcriptase polymerase chain reaction(RT-PCR).The activity of caspase-3 was detected by chemiluminescence assay. After exposing to 10μmol L−1 and higher concentration of VK2, it could inhibit MUTZ-1 cells proliferation in a dose-and time-dependent manner(p&lt;0.05). At concentration of 5μmol/l VK2 treatment, it might accelerate cellular proliferation, but there’s no significant difference compared with control group. Apoptosis peak on FCM and positive Annexin-V FITC/PI on cell membrane showed that VK2 induced apoptosis of MUTZ-1 cells in a dose-and-time-dependent manner, G0/G1 cell cycle arrest, significantly dow-regulated the expression of bcl-2 and survivin, but had no effect on the expression of bax.The activities of caspase-3 were significantly increased. Low concentration of VK2 could facilitate cell proliferation. The higher concentration of VK2 could induce apoptosis of MUTZ-1 cells. These results indicate that VK2 induces MUTZ-1 cells apoptosis by activating caspase-3 pathway, the apoptosis related genes bcl-2, survivin down-regulated might play an important role in this process.

scholarly journals Anti-tumor effects of isoliquiritigenin in Bcl-2/Bax and PCNA expression of T24 human bladder cancer cells

2020 ◽  
Author(s):  
Zhanbin Huang ◽  
Qianmei Wu ◽  
Zhong Wang
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
Wound Healing ◽  
Western Blot ◽  
B Cell Lymphoma ◽  
Nuclear Antigen ◽  
Control Group ◽  
Dependent Manner ◽  
Wound Healing Assay ◽  
T24 Cells

IntroductionThe aim of this study was to investigate the proliferation and apoptosis of T24 after treatment with isoliquiritigenin (ISL) and to explore the underlying mechanism.Material and methodsT24 cells were divided into 6 groups and cultured. The five experimental groups were seeded into medium with 10, 20, 40, 80, and 160 µM ISL and the control group was administered with 0.01% DMSO. The cell morphology was observed using an optical microscope. The proliferation of cells and half maximal inhibitory concentration (IC50) were determined and calculated by MTS. Cell cycle and apoptosis were evaluated by flow cytometry. The migration ability of cells was detected by wound healing assay. The expression of B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), and proliferating cell nuclear antigen (PCNA) was determined by RT-qPCR and western blot.ResultsAt 24 h after treatment with ISL, morphology revealed that cell number diminished and the cell volume shrank with increasing concentration. According to the results of MTS, ISL significantly inhibited the T24 cell proliferation in a dose- and time-dependent manner. Flow cytometry showed that ISL could block the replication of T24 cell DNA from S phase to G2, therefore promoting cell apoptosis. Wound healing assay showed that at 48 h after treatment with ISL, the migration of T24 cells was remarkably inhibited. The results of RT-qPCR and Western blot revealed that after 24 h of treatment, the expression levels of Bcl-2 and PCNA were down-regulated, and the expression level of Bax was increased in the different dose of the ISL treated group.ConclusionsISL possesses detrimental effects on the viability of BC cell T24 in a dose-dependent manner via blocking the cell cycle and inducing apoptosis by down-regulation of PCNA and Bcl-2 expression and up-regulation of Bax expression.

An Overview of the Application of Poly(lactic-co-glycolic) Acid (PLGA)-Based Scaffold for Drug Delivery in Cartilage Tissue Engineering

2021 ◽  
Author(s):  
Majid Pourentezari ◽  
Hengameh Dortaj ◽  
Batool Hashemibeni ◽  
Maryam Yadegari ◽  
Abbas Shahedi
Keyword(s):  
Drug Delivery ◽  
Tissue Engineering ◽  
Glycolic Acid ◽  
Biomedical Application ◽  
Cartilage Tissue Engineering ◽  
Biological Properties ◽  
Cartilage Tissue ◽  
Plga Scaffold ◽  
Small Molecule Drugs ◽  
Synthetic Scaffold

Poly(lactic-co-glycolic) acid (PLGA) has attracted a considerable amount of interest for biomedical application due to its biocompatibility, tailored biodegradation rate (depending on the molecular weight and copolymer ratio), approval for clinical use in humans by the U.S. Food and Drug Administration (FDA), the potential to change surface properties to create better interaction with biological materials and being suitable for export to countries and cultures where planting products with animals is unusable. For commercial use and research, PLGA has been widely studied to control small molecule drugs, proteins, and other macromolecules. This study aims to review the studies that used PLGA scaffolding and its composites as a scaffold and drug delivery in cartilage tissue engineering. It is concluded from the results that the PLGA scaffold as a synthetic scaffold, when combined with natural scaffolds or hybrids, strengthens its biological properties and performs its function better.

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