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Objective@#To explore the biological characteristics of synovial fluid-derived mesenchymal stem cells (SF-MSCs) cultured in serum-free medium and the ability of in vitro reconstruction of three-dimensional cartilage combined with scaffold material.@*Methods@#Human SF-MSCs were cultured in serum medium and mesenchymal stem cells medium-serum free (MSCM-sf) respectively, then the proliferative ability and morphology of SF-MSCs were compared; The third passage SF-MSCs cultured in MSCM-sf were identified by flow cytometry, three-way(chondrogenic, osteogenic, adipogenic)differentiation assay and induced for chondrogenic differentiation when combined with polyglycolic acid/polylactic acid (PGA/PLA).@*Results@#SF-MSCs cultured in MSCM-sf had better morphology and proliferative ability than that cultured in serum medium. The expression levels of positive markers of the third passage SF-MSCs cultured in MSCM-sf, such as CD73 (99.5%), CD90 (98.9%) and CD105 (96.5%), were more than 95%. However, the overall negative markers (CD34, HLA-DR and CD11b) expressed less than 2%. Three-way differentiation staining was positive. The combination of SF-MSCs and PGA / PLA can be induced into cartilage in vitro.@*Conclusions@#SF-MSCs cultured in MSCM-sf can be amplified under the condition of maintaining the stem cell characteristics, and can be combined with PGA/PLA scaffold to construct three-dimensional cartilage in vitro.
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BACKGROUND:There were certain differences in the anatomical structure of knee joint between man and woman. Gender knee solution is a new type of artificial knee joint prosthesis, which is specialy designed for women. Theoreticaly, the outcome of unisex total knee arthroplasty prosthesis should be related to gender, but we did not find very obvious differences in practical clinical work. OBJECTIVE:To analyze the differences in curative effects of the unisex knee arthroplasty prosthesis between male and female patients undergoing total knee arthroplasty so as to find out if it is necessary to apply female knee prosthesis among appropriate crowd. METHODS: We retrospectively analyzed the clinical data of patients undergoing total knee arthroplasty from May 2001 to June 2011. Among 312 patients (350 knees receiving total knee arthroplasty), patients who died within 3 years after surgery, lost to folow-up and underwent revision were excluded. Changes in knee functions and imaging were observed between males and females. RESULTS AND CONCLUSION:The women and men had similar mean pre-operative knee scores, flexion function, pain score among 285 patients (300 knees). However, the women had significantly lower mean extension function and function scores than the men. There were no significant differences in improvement in the knee scores, flexion, the pain and knee function between women and men before and after surgery. Nevertheless, men had better extension than women. No significant difference in radiolucencies and complications was seen between females and males. Therefore, there was little difference in outcomes between women and men who used the unisex total knee arthroplasty prosthesis.
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Objective To establish a simple and effective method for isolation and culture of mouse adipose-derived stem cells (mASCs)in vitro,in order to provide the sufficient sources of seed cells for the research of mesenchymal stem cells.Methods The mouse inguinal fat tissues were isolated in vitro and performed a digestion with 0.1% collagenase type NB4,then adipose-derived stem cells(ASCs)were seeded and adhered to the culture dishes in low glucose DMEM containing 10% fetal calf serum.The cellu-lar morphology,in vitro proliferation capacity,multidifferentiation potential and immunophenotype were assessed.Results The mASCs showed good cell morphology,extremely strong proliferation capacity and potential of adipogenesis,osteogenesis and chon-drogenesis via in vitro three-dimensional induction.The cellular surface antigen phenotype was consistent with that reported by lit-erature,and the expression of CD34 and CD105 was positive,Sca-1 was highly expressed,CD45 and SSEA-1 were not expressed. Conclusion Using the experimental methods in this research can culture the high purity of mASCs with the excellent stem cell properties and extremely strong proliferative ability.
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Cartilage is one of the earliest reconstructed tissues used in tissue engineering. Due to the lack of appropriate seeding cells, cartilage tissue engineering is, however, relatively lagged behind. With the emergence of stem cell research, adipose stem cells(ASCs) are introduced as seeding cells into tissue engineering for possessing many advantages such as wide spreading, large amount of cells available and easy to obtain. However, the outcome of tissue engineered cartilage construction by ASCs is not as ideal as that by bone marrow stem cells (BMSCs) yet. Low efficiency of ASC chondrogenesis is considered the major cause. This review summarizes the purification of adipose-derived cells, maintenance of sternness and optimization of ehondrogenie induction, which play vital roles in improving ASC s chondrogenesis.
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@#Objective To investigate the action of chondrogenesis differentiation of bone marrow stromal cells (BMSCs) transfected with adeno-hTGF-β1. Methods In the experiment group, replication-deficient a denoviruses carrying human hTGF-β1 complementary DNA (adeno-hTGF-β1 was constructed and applied to transfect to the first generation BMSCs. As a control, each BMSCs was transduced with 200 pfu of adeno-LacZ gene. One day after transfer, BMSCs were trypsinized, counted, and 5×105 cells aliuots were spun down at 500 rpm per minute in 15 ml polypropylene conical tubes and then cultured in a defined medium in an incubator at 37℃ for 21 days. The aggregates were harvested at time points to 21 days and assessed by gross observation, histological analyses and immunohistochemical localization of type Ⅱ collagen. Results When harvested at 21 days, each pellet shrinked to spheroid tissue with apearly opalescence in gross morphology and found to be relatively firm. H.E staining showed elongate dlining cells appeared as perichon drium-like cells at the surface. Some nests of cartilage were observed at the substrate of the tissue. Mature chon drocytes were embeded in the lacuna in the experiment group. In addition, Safranin'O staining confirmed the presence of sulfated proteoglycans in the ECM of chondrogenesis region. Immunohistochemical staining revealed the presence of type Ⅱ collagen in chondrogenesis region. By contrast, HE staining showed no evidence of cartilage formation in the control group. They were fibrous tissue with no architectural feature. Safranin'O staining and Immunohistochemical staining showed no evidence of sulfated proteoglycans or typeⅡ collagen expression. Conclusion BMSCs transfected with adeno-hTGF-β1 could induce its chondro-genesis when aggregate cultured in a defined medium in vitro, laying a foundation for the application of hTGFβ1 gene-transfected BMSCs in cartilage tissue engineering.
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In vitro construction technology is a key approach to industrialization and clinic application of engineered cartilage. However, it is very difficult to acquire a functional engineered cartilage with the present technology. Bioreactors can simulate the cartilage microenvironment in vivo and are expected to make up the shortcoming of the present technology. Current bioreactors in use are designed according to fluid shear pressure, hydrostatic pressure and/or direct compression, all of which can promote the development and mature of cartilage in vivo. Due to the failure to achieve ideal results by a single-purpose bioreactor, it will become a development direction in future to design and produce a compound bioreactor. This article reviewed the advances in the bioreactor for cartilage tissue engineering.