The Experimental Research Of OA Kneepad In Preventing And Treating Knee Osteoarthritis

Objective To develop an OA kneepad and explore its mechanism and efficacy on the prevention and treatment of osteoarthritis of the knee through observing the effects of OA kneepad on cytokines,matrix-dissolution,hyaluronic acid,oxygen free radical metabolism, articular cartilage micro-structural changes,cell proliferation and apoptosis.Methods1 Development of OA kneepad:Under the principle of"treating cold syndrome with hot-natured drugs" and "treating blood stasis syndrome with promoting blood circulation and removing blood stasis drugs",PIC16F630 SCM was served as the core and mainly composed of four parts including the boost circuit,the output,single-chip microcomputer control,as well as electric soft membrane.Dynamic range and frequency inverter dredging wave of treatment was carried out by settings of SCM via different keyboards.The instrument will be automatically shutdown 30 min later owing to the interruption of internal timer of SCM.2 Experimental Research:Sixty Japan rabbits were randomly divided into two groups, namely:the normal group(10) and surgery group(50).Knee osteoarthritis models were made by using the method of modified Hulth and then randomly divided into five groups,namely, model group and the control group(microwave),experimental group 1(electricity), experimental group 2(thermal),the experimental group 3(kneepad),with corresponding treatment.After 8 weeks,the IL-1β,IL-6,TNF-αand MMP-3,HA concentrations in articular synovial fluid and MDA,SOD,NO concentrations in blood were measured respectively.After 16 weeks,the IL-1β,IL-6,TNF-αand MMP-3,HA concentrations in articular synovial fluid and MDA,SOD,NO concentrations in both blood and synovial were also measured respectively.Besides,tissue morphology were analyzed by using CR tablets,light microscopy, transmission electron microscopy,Tunel,PCNA,and relative mRNA expression levels of Bcl-2 and P53 in articular cartilage cells were detected by real-time quantitative RT-PCR.Results1 Development of OA kneepad:The kneepad density-wave output can be controlled by 2Hz different frequencies to 300 Hz stimulation pulse component,the pulse width of 100 to 150μs between the positive and negative output symmetry to the stimulation pulse.The rechargeable mobile phone batteries(3.7V) were served as power supply and SCM as timer with automatic shutdown after 30 minutes.The temperature of electric soft membrane was controlled in 40℃.2 Experimental Research:IL-1β,IL-6,TNF-αand MMP-3 in synovial fluid,MDA,NO in blood concentrations of model group,the control group,the experimental group 1,group 2,group 3 were high expression levels,16 weeks later was significantly higher than those in 8 weeks,The expression of the HA in synovial fluid and SOD in blood concentration was significantly lower than those in 8 weeks,16 weeks later,MDA and NO concentration of synovial,P53 relative mRNA in articular cartilage cells,the expression level of AI were overexpressed,and SOD concentration,Bcl-2 mRNA expression levels of articular cartilage cells,PI were low expression,and the normal group was significant difference(P＜0.01,P＜0.05).8 weeks later,IL-1β,IL-6,TNF-α,MMP-3 and HA in synovial fluid,MDA,NO, SOD in blood,in the control group,the experimental group 1,group 2,group 3 and Model group there were significant differences(P＜0.01,P＜0.05),there were significant differences (P＜0.05) in the control group,experimental group 1,group 2 and the experimental group.16 weeks later,IL-1β,IL-6,TNF-αand MMP-3,HA in synovial fluid,MDA,NO,SOD in blood and synovial,Bcl-2,P53 mRNA expression in content of articular cartilage cells,AI,PI, there were significant differences(P＜0.01,P＜0.05)in the control group,experimental group 1, group 2,group 3 and model group;In the control group,experimental group 1,group 2 and group 3 there were significant differences(P＜0.05).In 16 weeks and 8 weeks,IL-1β,IL-6, TNF-αand MMP-3,HA in synovial fluid,MDA,NO,SOD in blood in group 3,there was significant difference(P＜0.05).The report of CR corresponds to the sequentiae condition.In the normal group,joint space is normal,and the articular facet is complete and slick with no osteophyte;In the model group,the is obviously narrowing between the arthral wall blank with the obvious osteophyte.The light microscope corresponds to the sequentiae condition.In normal group,the four layers'structures of arthrodial cartilage are clear,distinguishing.The cartilage cells line up in order with the shape of pillar,the disposition of caryotin uniformity,and the caryon is clear,In the model group,the layer of arthrodial cartilage gets to thinningz.In some districts,cartilage cell shows karyopyknosis and cellular necrosis,In the calcifying layer,cartilage cells line up in disorder and four layers'structures are uneasy to tell the differences.In the normal group,Electron microscopy showed normal cartilage cells were Oval,cells and cell memrane are integrity.In intracytoplasm,there are abundant rough endoplasmicsreticulum(PER),dictyosome,The caryon is integrity,and the disposition of caryotin is uniformity;In the model group,cartilage cells obviously pyknosis and their shape shows irregularity.The areolae around cells disappear.In intracytoplasm,cell organs coagulate to falps which show electron dense and it is uneasy to tell the differences.The shape of caryon shows irregularity,and caryotin gathers densely while spalling in caryon.In GroupⅢ,there is narrowing between the arthral wall blank.There is the little osteoplyte at the verge of the articulus and the articular facet is obviously rough and deformation.Cartilage cell shrinks back slightly.Intranuclear different dye-substance is increasing,it is thus clear that there are lipid droplet and microfilament in the intracytoplasm.Changes of morphological organization of control group,experimental group 1 and group 2 between model group and experimental group3.Immunohistochemical study results:normal group,group 3,group 1,group 2,control group and model group,TUNEL-positive rate increased gradually,PCNA-positive rate decreased gradually.Conclusion1 The OA kneepad with PIC16F630 microcontroller core can control the output signal frequency,pulse width,intensity and work time by controlling keyboardbuttons.2 By using low-frequency dynamic frequency density wave,the OA kneepad could regulate many aspects of pathological process of knee OA including:inhibition of biological effects of cytokine,MMP-3,HA,oxygen free radicals and NO;enhancing the biological activity of SOD;up-regulating the mRNA expression of Bcl-2 as well as down-regulating the mRNA expression of P53,thereby to inhibit the apoptosis of cartilage cells and delay the cartilage degeneration of the macro-morphology,cartilage cells and cartilage matrix.