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.

Autologous cells of bone, cartilage, intervertebral disc and tendon, are hard to be cultured and proliferate in vitro and thus have difficulty to be introduced in the repair of damaged tissue. Stem cells have the ability to self-renew and differentiate into many tissue types. Recent progress in stem cell research has led to an enthusiastic effort to utilize stem cells for orthopaedic tissue regeneration. Due to the abundance and easiness of harvest, adipose-derived mesenchymal celI(ASC) is an attractive, readily available aduh stem cell that has become increasingly popular for use in stem cell and tissue engineering applications. This review focuses on the use of ASC in orthopaedic tissue repair. Recent results from in vivo defect repair utilizing ASC suggested the great potential of ACS in clinical orthopaedic tissue regeneration.

Objective This experiment aimed to confirm whether man made porous chitosan scaffold is a appropriate scaffold for chondrocyte culutre of tissue engineering Methods Chondrocytes were seeded onto porous chitosan and chitosan collagen complex scaffolds for culture in a three dimensional environment The scaffolds in hydrophilia and adhesion to chondrocytes were observed with light microscope and scanning electron microscope The number of the cells attached to the scaffolds and the function of the cells were detected with MTT automated colormetric microassay Result Chondrocytes can multiple and secrete the matrix on the poros chitosan and chitosan collagensc scaffolds The cell adhesion rates were 81 25% and 87 50% respectively Conclusion Chitosan can be fabricated into a suitable three dimensional porous scaffold Porous chitosan collagen complex scafflold may be a more suitable scaffold for chondrocyte culutre of tissue engineering

OBJECTIVETo compare the tissue engineered cartilage constructed with chondrocytes derived from auricular and articular cartilage.

METHODSChondrocytes were isolated from porcine auricular and articular cartilage, and BMSCs were obtained from bone marrow aspirate and cultured. Each kind of chondrocytes were resuspended alone or mixed with BMSCs at a ratio of 1:1, and seeded onto PGA/PLA scaffolds to construct tissue engineered cartilage (n = 4). The constructs were cultured for 8 weeks in vitro and then subcutaneously implanted into nude mice for 6 weeks. The differences between chondrocytes monoculture from articular and auricular cartilage or between each of them co-cultured with BMSCs were evaluated by gross view, measurement of thickness and wet weight, histological examinations including H&E, Safranin O, type II collagen, and Ponceau's & Victoria blue staining, and gene expression analysis of cartilage related genes.

RESULTSNo obvious differences were found histologically among the complexes constructed in vitro 8 weeks except for few elastic fibers secreted in the auricular chondrocytes + BMSCs co-culture group. Neo-cartilage is thicker in the groups of articular chondrocytes (38. 1% than the group of auricular chondrocytes, P < 0.05) and articular chondrocytes + BMSCs co-culture (19.3% than the group of auricular chondrocytes + BMSCs, P < 0.05). However, after 6 weeks in vivo the elastic fibers were found positive in the complexes constructed by auricular chondrocytes, and its staining was even stronger and more homogenous in the group of auricular chondrocytes + BMSCs co-culture. The tissues generated by articular chondrocytes alone and co-cultured with BMSCs both formed the characteristic features of three-layer structure of hyaline cartilage and ossified in vivo with significant up-regulation of COL10A1 and MMP-13. To summarize, auricular chondrocytes formed the elastic cartilage while articular chondrocytes formed the hyaline cartilage during the development of tissue engineered cartilage either by monoculture or the co-culture with BMSCs.

CONCLUSIONSThe chondrogenic response of chondrocytes from different cartilage origins demonstrates that an initial chondrocyte and cartilage type recapitulates the same in later tissue-engineered development.

Animals ; Bone Marrow Cells ; cytology ; Cartilage, Articular ; cytology ; Cells, Cultured ; Chondrocytes ; Coculture Techniques ; Ear Auricle ; cytology ; Mesenchymal Stromal Cells ; cytology ; Mice, Nude ; Swine ; Tissue Engineering ; methods ; Tissue Scaffolds

BACKGROUND: The transplantation of allogeneic cartilage has local immunological rejection, and it is necessary to further reduce the rejection to promote its application in clinic, thus it is significant to perform a series of experiments to induce local immune privilege.OBJECTIVE: To observe the in vivo growth of tissue engineered allogeneic cartilage reconstructed by chondrocytes transfected with recombinant retroviral vector pLNCX2-FasL.DESIGN: A randomized controlled observation.SETTING: Shanghai Jiao Tong University.MATERIALS: Thirty-six allogeneic New Zealand rabbits as recipients and 45 1-week-old chinchillas as donors, either sex,were purchased by the experimental animal center of Chinese Academy of Sciences. Amphotropic recombinant retrovirus coated cell line PT67 was purchased from Clontech Company; Dulbecco's modified eagle medium (DMEM), fetal bovine serum (FBS), G418 and Polybrene were bought from GIBCO BRL.METHODS: The experiment was carried out in original Shanghai Second Medical University from January 2000 to July 2005. The New Zealand rabbits were randomly divided into three groups: FasL-transfected group (n =12), untransfected group (n =12) and blank control group (n =12). The rabbit allogeneic cartilages were constructed by the compound of pLNCX2-FasL transfected chondrocytes and tissue engineered material of pluronic F-127. ① Gross observation and mass changes of the grafts: Corresponding materials were infused subcutaneously, the grafts were removed at 1, 2 and 3 months after transplantation for gross observation and the mass changes. ② Staining observation: The grafts were removed at 1, 2 and 3 months after transplantation, then prepared into sections, and observed by hematoxylin and eosin (HE), Safranin'O and Masson's trichrome stainings. ③ Antibody detection: Blood samples (1 mL) were collected at 1 and 2 months after transplantation, the chondrocytes of the chinchillas were lysed freezingly with lysis antigen as the mixed antigen, and separated by electrophoresis in agarose medium, then acted with serum of recipient to observe whether corresponding antibody generated. ④ Complement dependent cytotoxicity (CDC) test: The chondrocytes of chinchillas were prepared into cell suspension (2×109/L), and then seeded into 96-well plate, attached grew for 24 hours, then recipient serum was added for the CDC test, and the percentage of apoptotic cells was counted under microscope.MAIN OUTCOME MEASURES: ① Gross observation and mass changes of the grafts;② Histological changes; ③ Results of the antibody detection; ④ Percentage of apoptotic cells.RESULTS: All the 81 rabbits were involved in the analysis of results. ① Gross observation and mass changes of the grafts: Two weeks after inoculation, there were obvious nod formations at the inoculated sites, but no nod formed in the blank control group. The new cartilage tissues became smaller gradually and completely disappeared at 4 months in the untransfected group, whereas those in the FasL-transfected group became smaller, but still existed after 7 months. The masses of grafts in the FasL-transfected group were higher than those in the untransfected group (P ＜ 0.05). ②Histological observation: Plenty of lymphocytic infiltrations around cartilage tissue could be observed in the untransfected group, and obviously decreased in the FasL-transfected group. No lymphocyte was observed inside the chondrocytes.Masson's trichrome staining was performed, and it was observed under light microscope that the small white parts in the middle were immature chondrocytes, and there were green collagen around most of the mature chondrocytes. Safranin O staining showed strong positive reaction, suggested that there were rich glycosaminoglycan in matrix. ③ Antibody detection: The chondrocytes of the chinchillas were lysed freezingly with lysis antigen as the mixed antigen, then acted with serum of recipient, and the results showed that no corresponding antibody generated. ④ Percentage of apoptotic cells: The percentages of serum CDC apoptotic cells in the FasL can ransfected group, untransfected group and blank control group were 5%, 6% and 1%, which were all negative.CONCLUSION: Rabbit allogeneic chondrocytes transfected with recombinant retroviral vector pLNCX2-FasL can reconstruct tissue engineered cartilage, and can postpone the degeneration by 3 months.

0.05),between the two groups,but in body mass index [(24.451?3.752) vs(22.468?2.434),P=0.030],previous history of pancreatitis(23.8% vs.7.5%,P=0.046) and bloody ascites(95.24% vs.46.24%,P0.05).MODS rate was higher in HL-group(52.4% vs.29%,P=0.04).The two groups had similar operation rates(66.7% vs 80.6%,P=0.178),hospitalization days [(25.476?14.383) vs(22.796?7.191),P=0.216] and mortality rate(28.6% vs.11.8%,P=0.069).Conclusions Hyperlipidemic severe acute pancreatitis has the characters of heavier weight habitus,history of frequent recurrence,high incidence of bloody ascites and prone to develop MOF.

Objective To evaluate the correlation between the expression level of Nanog gene and clinical outcomes of GP regimen in the advanced non-small cell lung cancer (NSCLC).Methods 62 patients of NSCLC were treated by GP method,and the outcomes were investiged between Nanog positive and nagetive patients.The expression level of Nanong was evaluated by RT-PCR and immunohistology.Results 30 out of 62 patients (48.4 ％) were Nanog positive,9 patients (28.1％) were Nanog positive,and 23 out of 32 patients were Nanog negative (71.9 ％) who have the positive effect (CR+PR).However,among 32 treatment nagetive cases,there were 21 cases (70.0 ％) who were Nanog positive and 9 cases (30.0 ％) were Nanog negatve.Survival analysis showed that 5-years lifetime of Nanog positive patients was shorter than Nanong nagetive patients.Conclusion Nanog overexpression decreases the sensitivity of GP regimen and lifetime of NSCLC patient.Nanog expression level may provide a useful factor for clinical treatment and prognosis of NSCLC patient.

Objective To investigate the preliminary experience of mechanical thrombectomy with a tri-axial system of the Solitaire AB stent through a Neuro delivery catheter to treat intracranial large artery occlusion. Methods A tri-axial system was used to deliver the Solitaire AB stent through a Neuro delivery catheter to provide intracranial aspiration in close proximity to the stent. This technique was used in 1 case of acute middle cerebral artery occlusion and 1 case of acute basilar artery occlusion. Results Successful revascularization was achieved in these 2 cases. Thrombolysis in cerebral infarction (TICI)score was 3. The clot length of acute middle cerebral artery occlusion was 3 cm and the modified Rankin Scale (mRS)score of this case was 3 at 90 days follow-up. Another patient with acute bilateral vertebral occlusion was revealed successful recanalization by angiography. Conclusion The results suggest that this technique of a tri-axial system used of the Solitaire stent through a Neuro delivery catheter can effectively retrieve clots from the occlusive artery and minimize the chance of antegrade blood flow dislodging the thrombus.

BACKGROUND: There are plentful studies about bioreactor of tissue engineering of blood vessel, tendon, cartilage, heart valve,trachea, bladder and stern cell. OBJECTIVE: To construct small-sized tissue-engineerad blood vessels with human bone marrow stromal stem cells (hBMSCs) in improved bioreactor system.DESIGN, TIME AND SETTING: The single sample observation stuay was performed at the School of Mechanical and Power Engineering, East China University of Science&Technology, and Shanghai Tissue Engineering Research & Development Center from June 2005 to March 2008.MATERIALS: Vessel bioreactor was self-made by East China University of Science&Technology. hBMSCs were harvested from healthy volunteers. METHODS: A set of support bracket constructing tissue engineered blood vessels with the diameter of 2 mm was designed with the application of Finite Element Methods as an analysis method analyzing support bracket of tissue engineered small-sized blood vessel. Primary hBMSCs were first Collected and further cultivated exvivo. The third passage cultured cells were then seeded on the polyglycolic acid (PGA) to fabricate the cell-scaffold composite. Subsequently, this composite was subjected to dynamical culture in the blood vessel bioreactor. After cultured for 4 weeks, the composite was removed from the bioreactor.MAIN OUTCOME MEASURES: The following mentioned references were measured: composite growth; other correlation detection of the hBMSCs-PGA composite, RESULTS: Gross observation and scanning electron microscope were used at4 weeks after hBMSCs-PGA composite culture. It was observed that the tissue-engineered blood vessel had a bright color and certain elasticity. The blood vessel could rebound to its odginal shape after repeated press by the forceps.The secreted collagen matrix arrayed orderly around the cells and smooth muscle elastic acttn could also be detected in the formed tissues using immunohistochemistry. CONCLUSION: The in vivo mechanics conditions of blood vessels can be simulated using the current blood vessel bioreactor system. Using hBMSCs, the construction of tissue engineered small-sized blood vessels can be successfully achieved.

BACKGROUND: Classic isolation method of bone marrow mesenchymal stem cells (BMSCs) is Percoll density gradient centrifugation. Blood cell component was removed. However, this method is complicated. Preparation density was needed when isolating dog bone marrow. Moreover, centrifugation was frequent, which had a great damage to cells. OBJECTIVE: To establish methods of the isolation, proliferation, culture and osteoinduction of canine BMSCs, and observe the in vitro proliferation and ability to osteoinduction differentiation. METHODS: 10 mL bone marrow was extracted from dog posterior superior iliac spine, heparin anticoagulation, diluted using Hanks juice, treated with 1.077 g/mL Ficoll solution 3 mL, and centrifuged at 2 000 r/min for 20 minutes. Karyocytes were absorbed to form white cloudlike layering interface, and then centrifuged twice using DMEM supplemented with fetal bovine serum, incubated at 12×10~4/cm~2 at 37 ℃ in a 5% CO_2 incubator. Following subculture, cells were incubated in DMEM containing dexamethasone, β-sodium phosphoglycerol and ascorbic acid 2-phosphate. Immunocytochemical staining and immunofluorescence staining were utilized to detect osteocalcin, osteopontin and type Ⅰ collagen expression in osteoblasts. Alkaline phosphatase staining and alizarin red staining were performed. RESULTS AND CONCLUSION: 1.077 g/mL Ficoll density gradient centrifugation was used to isolate karyocytes that were significant compared with Percoll solution. Obtained BMSCs had high purity, good growth and the mean doubling time was 24 hours. Following in vitro osteogenic incubation of dog BMSCs, osteocalcin, osteopontin and type Ⅰ collagen showed positive expression. Alkaline phosphatase staining demonstrated bluish-green cytoplasm. Alizarin red staining showed red nodes in extracellular matrix, and could differentiate into osteoblasts in vitro.