Effects of fibroblast growth factors 2 and low intensity pulsed ultrasound on the repair of knee articular cartilage in rabbits.

OBJECTIVE: To investigate the effect of low intensity ultrasound irradiation combine with fibroblast growth factors (FGF2) on the repair of the knee articular cartilage and to explore its mechanism in rabbit. MATERIALS AND METHODS: The model of the rabbit knee joint injury was established. 40 rabbits were divided into four groups, including control group, model group, FGF2 group and FGF2 + low intensity pulsed ultrasound group (FGF2 + LIPU). The knee joints of rabbits were taken at 4 and 8 weeks, respectively. Histopathological changes were detected by Hematoxylin and Eosin stain (HE) and evaluated by Wakitani score. The expression of FGF2 mRNA was detected by Real-time polymerase chain reaction (RT-PCR) and the levels of Collagen I and Collagen II protein were analyzed by Western blotting. RESULTS: In FGF2 group and FGF2 + LIPU group, it was found that the tissues of knee joint were gradually repaired following the change of time. Further, the recovery was better in FGF2 + LIPU group. Cartilage defect areas were filled with cartilage-like cells and the repair surface was fused with surrounding cartilage in FGF2 and FGF2 + LIPU groups. Wakitani scores were consistent with HE results. The expressions of FGF2 mRNA were higher in FGF2 and FGF2 + LIPU group than the model group. Western blotting results showed that the levels of Collagen I and Collagen II protein in FGF2 and FGF2 + LIPU groups were significantly increased compared with that in model group. CONCLUSIONS: FGF2 and LIPU combined application on the rabbit knees joint repair is better than FGF2 alone. FGF2 and LIPU combination can promote the synthesis and secretion of collagen in chondrocytes, promote the differentiation and maturation of chondrocytes during the repair of cartilage defects.

Immunoregulatory effect of neuronal-like cells in inducting differentiation of bone marrow mesenchymal stem cells.

OBJECTIVE: To evaluate the immune activity of bone marrow mesenchymal stem cells (BMSCs), and explore the biological characteristics and capabilities of BMSCs and the potential to be differentiated into neuronal cells in vitro. MATERIALS AND METHODS: The BMSCs were isolated and proliferated in vitro to generate the xenogeneic mixed lymphocyte reaction. Moreover, peripheral BMSCs (pBMSCs) were added according to different ratios, which methods were stated as follows: 1: Dulbecco's Modified Eagle Medium (DMEM) + 10% Fetal Bovine Serum (FBS) + 1 µmol/L all-trans-retinoic acid (ATRA) + 20 µg/L basic fibroblast growth factor (bFGF) + 20 µg/L epidermal growth factor (EGF); 2: DMEM + 2% dimethyl sulfoxide (DMSO) + 100 µmol/L butylated hydroxyanisole (BHA). The immunofluorescence and immunohistochemical staining were finally used to evaluate the differentiation capabilities of human BMSCs (hBMSCs) induced in neuronal cells. RESULTS: hBMSCs inhibited the lymphocyte proliferation in the mixed lymphocyte reaction (MLR) system at a proportional inhibition rate with additional numbers of stem cells. At hour 2 after culture with method 1, the plasma of hBMSCs shrank to nuclei and perinuclear bodies and was visualized under the light microscope. At hours 3-5, most of the hBMSCs formed neuron-like cells with total cell number unchanged. Afterward, the hBMSCs turned into bipolar or multipolar shaped cells and interconnected into a large network at Day 3. With immunofluorescence and immunohistochemical staining, 60-70% of the hBMSCs showed neurospecific enolase (NSE) positive and 45-50% glial fibrillary acidic protein (GFAP) positive while the Nestin-positive cells decreased to 3.4%. However, when cultured 2 hours with method 2, the most of the hBMSCs formed bipolar or multipolar shaped cells, then died after 48 hours. 40-50% NSE and 35-40% GFAP were positively expressed. Significantly, the rate of Nestin-positive cells decreased from 63% to 1.6% from hour 2 after culture to hour 48. CONCLUSIONS: hBMSCs may be effective for cell therapy and tissue engineering for the capability of differentiating into neuronal-like cells, as well as the capability of inhibiting lymphocyte proliferation in MLR system.

[Clinical study on relationship of CD62p, CD63 and TSP to the qi-stagnation induced blood-stasis syndrome and qi-deficiency induced blood-stasis syndrome in aged patients].

OBJECTIVE: To observe the relationship of platelet activation to the Qi-stagnation induced blood-stasis (QSBS) or Qi-deficiency induced blood-stasis (QDBS) syndrome. METHODS: Expressions of platelet activating molecules, including alpha-granule membrane glycoprotein (CD62p), lysosomal integral membrane protein (CD63) and thrombospondin (TSP), in patients with QSBS and QDBS were determined quantitatively with flow-cytometry and specific monoclonal antibody against activated platelet. And platelet aggregation was tested simultaneously. RESULTS: CD62p, CD63 and TSP expressions in Blood-Stasis patients, both QSBS and QDBS, were higher than those in the normal control significantly (all P < 0.01); all the three expressions were higher in QSBS group than those in QDBS group (all P < 0.01), Positive correlation was shown between CD62p and CD63 (r = 0.740, P < 0.01), CD62p and TSP (r = 0.744, P < 0.01), TSP and CD63 (r = 0.635, P < 0.01), and between CD62p and ADP induced platelet aggregation (r = 0.715, P < 0.01). CONCLUSION: Platelet activation was involved in the pathogenesis and development of Blood-Stasis Syndrome, especially closely related with the QSBS Syndrome.