| Objective:1. Through animal experiment,we observed the influence on knee function, cartilage histology and cartilage extracellular matrix of rats with varying degrees of knee osteoarthritis (Knee osteoarthritis KOA) with Tongluozhitong medicine by low-frequency ultrasound transdermal penetration,and described the mechanism of action.2. Patients with varying degrees of knee osteoarthritis were therapied by Tongluozhitong medicine through low-frequency ultrasound transdermal penetration.We observed the changes of biomakers of cartilage extracellular matrix,VAS score,HSS function score and the Chinese version of Easy Scale Health Survey (SF-36), and investgated the clinical efficacy and mechanisms of the external treatment.Method:1. Animal experiment:rat models of osteoarthritis were prepared by cutting anterior cruciate ligament,;Left knees of rats were modeling while right knees were control.84male SD rats were randomly divided into three groups:control group(18), the group of ultrasonic transdermal penetration(33) and herbal topical groups(33).All rats were prepared and detected about the behavior by recording Lequesne MG score,and CTX-Ⅱ,COMP and MMP-3in serum were detected by ELISA.The control group were normal feeding,and were randomly selected to observe changes of behavior and CTX-Ⅱ,COMP and MMP-3at2/4/8weeks after surgery.After killed,the control group were observed on General observation of knee cartilage,and synovium and medial femoral condylar cartilage morphology and were evaluated by cartilage modified Mankin’s score and determine the degree of rats on KOA. Ultrasonic transdermal penetration group and herbal topical group were randomly selected for intervention treatment at the2/4/8weeks after surgery. Ultrasonic transdermal penetration group was transparentted with Tongluozhitong herbal to The operating knee by ultrasound and conductivity meter while the herbs topical group were just intervened by topical herbal.1/d,30min,2weeks.2weeks after death, the data was recorded which was as same as the control group.2.Clinical trials:80cases of knee osteoarthritis in line with western medicine and TCM syndrome diagnostic criteria of the blood stasis syndrome were divided into four categories,excellent, good, fair and poor according to the severity of HSS score,and each category was20patients and randomly divided into2groups:group of transdermal penetration of Chinese medicine and group of transdermal penetration of saline,10patients in each group.All patients took Chinese medicine Panlong seven basis to get Huoxuetongluo therapy. The group of transdermal penetration of Chinese medicine were therapied by Tongluozhitong medicine through ultrasound penetration while The group of transdermal penetration of saline were therapied by saline through ultrasound penetration,4times/week,30min,4weeks of continuous treatment. CTX-Ⅱ, COMP and MMP-3in serum of patients were measured by ELISA and VAS score, HSS score, and SF-36scores were collected before and after the treatment, to evaluate the clinical efficacy of the various groups.Results:1. Animal experiment1.1Behavior and Lequesne MG scoreBehavior control group changed obviously at2,4,8week after modeling,and Lequesne MG score increasted gradually.Ultrasonic transdermal penetration group compared with the control group, Lequesne MG score decreased at2,4and8week, there was significant difference (P<0.05); compared with the herbs topical group,Lequesne MG score decreased at2,4and8week, there was significant difference (P<0.05).1.2General observation of cartilageCartilage general observation of control group changed obviously,and the degree of cartilage degeneration aggravated.Ultrasonic transdermal penetration group compared with the control group, general observation of cartilage changed at2,4week,and the degree of cartilage degeneration eased;and there was no significant difference between Ultrasonic transdermal penetration group and the herbs topical group,1.3Cartilage tissue morphology and Mankin’s scoreCartilage tissue morphology of control group changed obviously at2,4,8week after modeling;Mankin’s score increasted and positively related to time.Ultrasonic transdermal penetration group compared with the control group, cartilage tissue morphology changed,and Mankin’s score decreased at2,4week.There was significant difference (P<0.01) at2weeks and general difference (P<0.05) at4weeks.Compared with the herbs topical group,Mankin’s score of Ultrasonic transdermal penetration group decreased sharply at2weeks,(P<0.05).1.4Synovial tissue morphologySynovial tissue morphology of control group changed obviously at2,4,8weekS.Ultrasonic transdermal penetration group compared with the control group and herbs topical group, Synovial tissue morphology changed at2,4,8week.1.5CTX-Ⅱ、COMP、MMP-31.5.1CTX-ⅡCompared with the control group,CTX-Ⅱ of ultrasonic transdermal penetration group decreased sharply at2,4weeks and there was significant difference (P<0.01),and it aslo decreased at8weeks (P<0.05).Compared with the herbs topical group,CTX-II of ultrasonic transdermal penetration group decreased at2,4weeks and there was significant difference at2,4,8weeks (P<0.05).1.5.2COMPCOMP of the control group increased gradually at2,4,8weeks.Compared with the control group,COMP of ultrasonic transdermal penetration group decreased sharply at2,4weeks and there was significant difference (P<0.01).Compared with the herbs topical group,COMP of ultrasonic transdermal penetration group decreased at4weeks (P <0.05).There was no difference between the control group and ultrasonic transdermal penetration group.1.5.3MMP-3Compared with the control group,MMP-3of ultrasonic transdermal penetration group decreased at2weeks and there was difference (P<0.05);compared with the control group,MMP-3of ultrasonic transdermal penetration group decreased sharply at4weeks(P <0.01). Compared with the herbs topical group,MMP-3of ultrasonic transdermal penetration group decreased at4weeks (P<0.05).At8weeks,MMP-3of ultrasonic transdermal penetration group was less than the control group and the herbs topical group,but there was no difference.2.Clinical results2.1HSS scoreIn patients rated as excellent, HSS score of transdermal penetration of Chinese medicine group increasted obviously,and there was significant difference(P<0.01),and the efficacy was better than group of transdermal penetration of saline(P<0.05).In patients rated as good, HSS score of transdermal penetration of Chinese medicine group increasted obviously,and there was significant difference compared to pretreatment.2.2VAS scoreIn patients rated as excellent,good and fair,VAS score of transdermal penetration of Chinese medicine group decreasted obviously after treatment,and there was significant difference compared to pretreatment.(P<0.05).VAS score of all patients in group of transdermal penetration of Chinese medicine increasted obviously after treatment,and there was significant difference compared to pretreatment.(P<0.01).2.3the MOS36item short from health survey, SF-36:In patients rated as excellent and good,the score of SF-36of transdermal penetration of Chinese medicine group on Physical Functioning,Role-Physical,Social Functioning and General Health increasted obviously after treatment.,and there was significant difference compared to pretreatment.(P<0.05). In patients rated as excellent, good and fair,The score of SF-36of transdermal penetration of Chinese medicine group on Bodily Pain and Mental Health increasted obviously after treatment.,and there was significant difference compared to pretreatment.(P<0.05).2.4CTX-Ⅱ、COMP、MMP-32.4.1CTX-Ⅱof transdermal penetration of Chinese medicine group decreasted after treatment;and in patients rated as excellent,good and fair,CTX-Ⅱ of transdermal penetration of Chinese medicine group decreasted after treatment,but there was no significant difference compared to pretreatment(P>0.05).2.4.2COMP of transdermal penetration of Chinese medicine group decreasted obviously after treatment,and there was significant difference compared to pretreatmentand (P<0.05).In patients rated as good and fair,COMP of transdermal penetration of Chinese medicine group decreasted after treatment,and there was significant difference compared to pretreatment (P<0.05);In patients rated as excellent,COMP of transdermal penetration of Chinese medicine group decreasted after treatment,but there was no significant difference compared to pretreatment(P>0.05).2.4.3MMP-3of transdermal penetration of Chinese medicine group decreasted obviously after treatment,and there was significant difference compared to pretreatmentand (P<0.05).In patients rated as excellent,COMP of transdermal penetration of Chinese medicine group decreasted after treatment,and there was significant difference compared to pretreatment (P<0.05).In patients rated as good and fair,MMP-3of transdermal penetration of Chinese medicine group decreasted after treatment,but there was no significant difference compared to pretreatment(P>0.05).Conclusion:1. Cutting anterior cruciate ligament of the knee can make biomechanical imbalance, cause cartilage degeneration and led to the occurrence of KOA.The degree of KOA in rat models through this approach has positive correlation with time.This kind of model can be well simulated to development of human KOA and this approach can cause different degrees of KOA models.2.Onset of effect to rat KOA models by Low-frequency ultrasound transdermal penetrating Tongluozhitong medicine may be slowing cartilage degeneration,especially slowing the degradation of the extracellular matrix of cartilage, reducing synovitis so that it can relieve knee inflammatory and improve limb function, which is better than ordinary topical method, but not obvious to severe knee osteoarthritis.3.low-frequency ultrasound transdermal penetration of Tongluozhitong medicine can reduce the CTX-Ⅱ, COMP and MMP-3levels in serum of patients with knee osteoarthritis, slow the degradation of extracellular matrix and thus slow down the degeneration of cartilage.It is able to significantly improve VAS score, HSS score and SF-36scores of patients, especially in mental health and physical pain, but with no significant effect to severe knee osteoarthritis. |
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