Shenzhen Key Laboratory of Tissue Engineering

Shenzhen, China

Shenzhen Key Laboratory of Tissue Engineering

Shenzhen, China

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Duan L.,Sun Yat Sen University | Duan L.,Shenzhen Key Laboratory of Tissue Engineering | Duan L.,Shenzhen Second Peoples Hospital | Ma B.,University of Zürich | And 9 more authors.
American Journal of Translational Research | Year: 2015

Abstract: Autologous chondrocyte implantation (ACI) is a golden treatment for large defects of the knee joint without osteoarthritis or other complications. Despite notable progresses, generation of a stable chondrocyte phenotype using progenitor cells remains a main obstacle for chondrocyte-based cartilage treatment. Monolayer chondrocyte expansion in vitro is accompanied by chondrocyte dedifferentiation, which produces a non-specific mechanically inferior extracellular matrix (ECM) unsuitable for ACI. In-depth understanding of the molecular events during chondrocyte dedifferentiation is required to maintain the capacity of in vitro expanded chondrocytes to produce hyaline cartilage-specific ECM. This review discusses key cytokines and signaling pathways involved in chondrocyte dedifferentiation from the standpoint of catabolism and anabolism. Some potential therapeutic strategies are also presented to counteract chondrocyte dedifferentiation for cell-based cartilage therapy. © 2015, E-Century Publishing Corporation. All right reserved.


He M.-J.,Anhui Medical University | He M.-J.,Shenzhen Second Peoples Hospital | Wang D.-P.,Anhui Medical University | Wang D.-P.,Shenzhen Second Peoples Hospital | And 5 more authors.
Chinese Journal of Tissue Engineering Research | Year: 2014

BACKGROUND: Low-temperature rapid prototyping technology is a new kind of rapid prototyping technology, and it is rapidly used in the preparation of bone tissue engineering scaffolds because it can make scaffold forming controllable and can keep the biological activity of the materials, also can easily realize the scaffold with porous of three-dimensional structure and other advantages. OBJECTIVE: To investigate the preparation process of polyethylene glycol-modified polylactic acid-glycolic acid/nano-hydroxyapatite (PLGA-PEG/n-HA) using the low-temperature rapid prototyping, and to test its performance. METHODS: PLGA-PEG/n-HA and PLGA/n-HA were prepared by low-temperature rapid prototyping equipment. Under an electron microscopy, we observed ultra-structure of the scaffolds. Immersion (ethanol) method was used to test the porosity, and electronic testing machine was used to determine the material mechanical properties. Then these two kinds of scaffolds with rat osteoblasts were cultured in vitro, the cell adhesion rate was detected by precipitation method after 12 hours, and cell counting kit-8 method was used to determine the cell proliferation at culture days 1, 3, 5, 7, 9, 12. RESULTS AND CONCLUSION: Both of the two scaffolds had ideal aperture range and high porosity. But the aperture range of PLGA-PEG/n-HA scaffolds had large fluctuations, and the average aperture was smaller than that of PLGA/n-HA. Some pores were closed up. The cell adhesion rate and the cell growth curve of PLGA-PEG/n-HA was better than that of PLGA/n-HA (P < 0.05), but the mechanical properties were less than PLGA/n-HA (P < 0.05). The results showed the PLGA-PEG/n-HA scaffolds had good cell compatibility.


Fu Z.-J.,Shantou University | Fu Z.-J.,Shenzhen Second Peoples Hospital | Zhang J.-F.,Shantou University | Zhang J.-F.,Shenzhen Second Peoples Hospital | And 19 more authors.
Chinese Journal of Tissue Engineering Research | Year: 2014

BACKGROUND: Human hypoxia-inducible factor-1 alpha can regulate the expression of osteogenic and angiogenic genes, and promote osteogenic activity. OBJECTIVE: To observe the expression of osteogenic genes in rat bone marrow mesenchymal stem cells carrying human hypoxia-inducible factor-1 alpha slow virus infection. METHODS: Hypoxia-inducible factor-1 alpha was obtained from Hela cells using RT-PCR. Lentivirus expression vector plasmid carrying hypoxia-inducible factor-1 alpha (Lenti-HIF-1α-eGFP) was constructed. 293Ta cells with LentiPac HIV mixed packaging plasmid was packaged, and then lentivirus was obtained. Rat bone marrow mesenchymal stem cells were isolated and cultured using direct whole bone marrow adherent method. Bone marrow mesenchymal stem cells were identified using flow cytometry. Bone marrow mesenchymal stem cells were infected with slow virus for 1, 4, 7 and 14 days. Bone morphogenetic protein-2, osteocalcin, osteopontin and alkaline phosphatase expression levels were detected in bone marrow mesenchymal stem cells using real-time fluorescent quantitative PCR. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells were effectively infected with Lenti-HIF-1α-eGFP. Real-time fluorescent quantitative PCR results revealed that bone morphogenetic protein-2, osteocalcin, osteopontin and alkaline phosphatase began to obviously overexpress from 4 days after infection with Lenti-HIF-1α-eGFP until 14 days. Results suggested that hypoxia-inducible factor-1 alpha could elevate the osteogenic activity of bone marrow mesenchymal stem cells.


Huang H.-Y.,Shantou University | Liu G.-P.,Second Peoples Hospital Of Shenzhen | Duan L.,Second Peoples Hospital Of Shenzhen | Duan L.,Shenzhen Key Laboratory of Tissue Engineering | And 6 more authors.
Chinese Journal of Tissue Engineering Research | Year: 2014

BACKGROUND: The umbilical cord blood is rich of mesenchymal stem cells, which can be used as a new source of seed cells in tissue engineering. OBJECTIVE: To compare two methods for culturing, expanding and purifying the mesenchymal stem cells in vitro isolated from the human umbilical cord blood. METHODS: The full-term birth cord blood of 40 cases was collected under sterile conditions with heparin anticoagulation. Ficoll density gradient centrifugation was used to isolate the mononuclear cells from the umbilical cord blood. The cases were randomly divided into two groups according to the different culture media. Twenty cases of umbilical cord blood were cultured in MesenGro human mesenchymal stem cells culture medium (group A), and the remaining 20 cases of umbilical cord blood were cultured in Dulbecco’s modified Eagle’s medium(group B). The occurrence time of fusiform mesenchymal stem cells and cell colony, culture time and the number of primary cells in the two groups were compared. Cells which grew well were selected to detect the surface markers by flow cytometry. RESULTS AND CONCLUSION: The mean occurrence time of fusiform mesenchymal stem cells and cell colony, culture time and number of primary cells in group A were better than those in group B (P < 0.01). The strong expression of the surface markers of mesenchymal stem cells (CD73 and CD105) was found by flow cytometry, of which the positive rate was 99.1%. No expression of the surface markers of hematopoietic stem cells (CD45 and CD34) was seen, of which the negative rate was 99.3%. The number, morphology, growth rate and culture time of umbilical cord blood mesenchymal stem cells cultured in MesenGro human mesenchymal stem cells culture medium were better than those cultured in Dulbecco’s modified Eagle’s medium. Cells cultured in MesenGro human mesenchymal stem cell culture medium can better express surface markers of mesenchymal stem cells. © 2014, Journal of Clinical Rehabilitative Tissue Engineering Research. All rights reserved.


Chen L.,Guangzhou Medical College | Chen L.,Second Peoples Hospital of Shenzhen | Chen L.,Shenzhen Key Laboratory of Tissue Engineering | Zhu W.-M.,Second Peoples Hospital of Shenzhen | And 19 more authors.
BioMed Research International | Year: 2013

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 scaffold in vitro and 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. © 2013 Lei Chen et al.

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