The Establishment And Treatment Of The Osteoporosis Model In Microfluidic Device

Objective: The occurrence of osteoporosis is inseparable with the decreasing of bone formation and the increasing of bone resorption. Osteoblasts, the basal cells of bone formation, those apoptosis and viability decrease are associated with the formation of osteoporosis. Glucocorticoid, as the common anti-inflammatory in clinic, is very harmful to skeletal system. This experiment is based on microfluidic technology, established the glucocorticoid-induced osteoporosis(GIOP) model and treat it simultaneously. Through the observation, we found the optimal induction concentration, also provided important foundation for the clinical study of osteoporosisMethods: The design and fabrication of chip was used soft lithography method by Dalian Institute of Chemical Physics. The material that fabricated the chip was Polydimethylsiloxane. PDMS and glass sheet were irreversible sealed by plasma treatment. We chose the MC3T3-E1 as the basal cells, after conducting osteogenetic differentiation induction, perfusing with drugs for 24 h with the speed of 0.1μl/min in chips, and the model was established. Then we detected the apoptosis level and the activity of alkaline phosphatase(ALP).We set three experiment groups: first, dexamethasone(Dex) perfusion group; second, Li Cl perfusion group; third, both dexamethasone and Li Cl perfusion group. Based on the feature of the chip, we calculated the drug concentration of every channel. Adopting the fluorescence staining method to analysis the apoptosis condition of induced osteoblasts. Using Image-pro Plus6.0 software to analysis the morphology, numbers, apoptosis situation and ALP activity.Processing statistical analysis by SPSS 16.0 software.Results:(1) The fabrication of microfluidic chip The microfluidic device in our study was mainly composed of an upstream concentration gradient generator(CGG) and a downstream cell culture module. The design of the CGG was based on the work previously presented by Jeon et al. As twostreams travel down in CGG,they are repeatedly split at the nodes, combined with neighboring streams in laminar fashion, and allowed to mix by diffusion in serpentine channels. Consequently, the solutions are continuously diluted and a series of concentrations are produced in the outlets of CGG. For the current study, our one solution(concentration 0) and the other one(concentration10μmol/L) were introduce d into CGG, the concentrations in the eight outlets are, respectively, 0, 2, 4, 6, 8and10μmol/L, according to the equation described by Jeon et al.The cell culture module is composed of an array of six cell culture chambers that are integrated with the CGG unit. The chambers were miniaturized: 1200 mm long, 1200 mm wide, and 150 mm height. The volume of each chamber is about 2μL.(2) The effect on osteoblasts proliferation and activity after perfusion with dexamethasone and Li Cl After perfusion with Dex, the morphology of osteoblasts changed from fiber-like to round-like, the proliferation and activity also had obviously decreasing(P<0.05),and the speed of proliferation decrease with the Dex concentration increase, to channel 4,the numbers of osteoblasts began to decrease.Meanwhile, after perfusion with Li Cl, the cells have no change.(3) The effect on osteoblasts apoptosis after perfusion with dexamethasone and Li Cl After perfusion with Dex,the osteoblasts appeared apoptosis, including the decreasing of mitochondrial membrane potential, the changing of nuclear morphometry and plasma membrane permeability(P<0.05),also with the concentration of Dex increased,the apoptosis turned up obviously. However, after perfusion with Li Cl, the cells have no change.(4)The effect on osteoblasts ALP activity after perfusion with dexamethasone and Li Cl The ALP activity of osteoblasts turned out decrease after perfusion with Dex(P<0.05),and with the concentration of Dex increase the ALP activity decrease. After perfusion with Li Cl, the cells have no change.Conclusion:Based on the microfluidic platform,we established the GIOP model using osteoblasts.We also have a deep study on the mechanism of osteoporosis and found the optimal concentration of Dex to form osteoporosis, which provide a novel platform for better clinical research and treatment of osteoporosis.Objective: To analyze and discuss the therapeutic effect of arthroscopy in treatment of synovial chondromatosis of the knee joint.Methods: We chosed sixteen patients of chondromatosis treated by our hospital randomly, from September 2010 to September 2013. All the patients were undergone probing, excising diseased synovium, removing loose bodies, and cleaning up degenerative tissues. Making a comparison of the therapeutic effect of arthroscopy between preoperation and postoperation.Results: The incision of all the sixteen patients healed well,and all of them were followed up by the time twelve months to twenty-four months(mean:16.75±3.6).The preoperative main symptoms all disappeared, joint function recovered well and joint motion was improved obviously. The knee joint Lysholm scale from 49.18±8.79 preoperatively to 83.75±4.9(P<0.05) postoperatively. Fifteen were cured and one turned better. And there is no severe complication and recurrence occurred.Conclusion: Arthroscopy is an effective way for the treatment of synovial chondromatosis of the knee joint, with advantages of minimal invasion, exhaustive excision of the diseased synovium and quick recovery. It’s worth clinical popularization and application.