Experiment And Clinical Research Of TongLuShengGu Capsule Combined With Principle Of Biomechanics On Treating Osteoarthritis

Osteoarthritis was a common desease in orthopedics.It severely puzzled people's life. At present there were only symptomatic treatment and supporting treatment on osteoarthritis. TongLuShengGu capsular combined with biomechanics principle was a effective method for OA which invested by professor yuanhao of gunzhou university of Traditional Chinese medicine. Our purpose was to study its effect and mechanism on treating OA.1 The effect of TongLuShengGu capsule on treating experiment osteoarthritis of rabbits1.1 Objective:To study treatment effect and mechanism of TongLuShengGu Capsules on treating osteoarthritis of rabbits, (caroid-induced osteoarthritis)1.2 Methods: sixty New Zealand rabbit were randomly divided into six groups, i. e. normal group, model group, zhuguguanjie group, there TongLuShengGu capsular group at different dosages .carotid were injected into rabbit knee joint for there times in one week. After this , administrated with TongLuShengGu capusular and ZhuGuGuanJie pill respectively for four weeks .After 4 weeks of treatment, macroscopic and microscopi of knee joint , cartilage biochemical and serum of phosphonium , Calcium, ACPase and AKP examination were done.1.2.1 articular macroscopic examinationCut the left knee joint , Observed knee joint by naked eye.1.2.2 blood biochemical test3 ml artery blood was took from rabbit ear before animal death.the contents of calcium, phosphonium , ACPase and AKP in serum was determined.1.2.3 cartilage biochemical testTaken cartilage from animal knee to make a biochemical test ?the water Content rate testSuck water from cartilage surface with the filter paper and then weight it, put cartilage in acetone for 24 hours and then dry cartilage under room temperature, weight it precisely again, computed water content rate, ?the glycuronic acid testTake 5mg cartilage specimen, placed in 5ml ampoule, add lml 2N HCL, deal with water bath for 2 hours in 100 degrees, and then determined glycuronic acid content by bitter' s methods . (3) hydroxyproline content testTaken 30mg cartilage specimen, determined hydroxyproline content by improvemental Neuman methods. 1.2.4 The cartilage pathologic examination ? light microscope observationFixed cartilage specimen with 10% formalin buffer, imbed with paraffin after decalcification, cut and than stained with HE . (2) Electron microscope observationFixed cartilage specimen with glutaral for 24 hours, washed with the 0.1M phosphoric acid buffer liquid for 3 times, fixed with 0.1% osmic acid for 90 minutes again, washed 3 times by deionized water to terminated osmic acid reaction, dehydrated in different density acetone , decalcificated with EDTA, soaked, imbed with the 61G ethoxyline resin, slendered semithin section after harden , cut into 0. 03 mm ultrathin section after allocation, observated after special electron stain . 1. 3 Result: 1. 3.1 articular macroscopic examinationIn normal group articular cartilage surface was smooth, had no tearing on cartilage surface and no synovium swollen , synovia was normal;In model group the cartilage became yellow , lose normal sheen, tear on surface, parts of cartilages contained defection and damaged, synovium was swollen and proliferated, observed inflamational adherence, synovia increased;In high dose group cartilage was smooth, the surface hadn't tearing and inflamation reaction on synovium, synovia was normal;In medium doses group, low doses group and medicine matched control parts of cartilages became yellow, teared on surface, had no inflamation and synovium proliferation, synovia was normal.1.3.2 blood biochemical analysisResults indicated TongLuShengGu capsular can influence the activity of ALP and ACPas . the content of calcium and phosphonium in serum has no markable different compared with all group.1.3.3 cartilage biochemical test(D Water content rate of model group was increased notably compared with thatof normalgroup. water contain rate of treated group and medicine controlgroup decresed notably compared with that of model group .? Glycuronic acid content of model group decreased markly compared^with thatof normal, glycuronic acid content of treated group and medicine control groupincreased nortably compared with that of model group.(Dhydroxyproline content measurement result show the hydroxyproline contenthad no obviously differenter in each group.1.3.4 cartilage pathologic observation?Light microscope observation In normal group cartilage surface was smooth, observed four layers in cartilage: superficial layer, migration layer, reflecting layer and calcify layer and no cell cluster phenomenon;reflecting layer cartilage cell arranged like pillar form;tide mark line was integrity;In model group cartilage surface was not smooth and had tear strip, defection and damage area, on surface layer cartilage cell disappeared, most of migration layer cell disappeared, fined cell cluster phenomenon, reflecting layer cell align like pillar form.The tide mark line was discontinuation;In high dose group cartilage surface was smooth, on surface layer chondrocyte disappear partly, at migration finede cell cluster phenomenon, the reflecting layer cell align like pillar form, the tide marks line was integrity;In medium dose group and matched control group cartilage surface had tear, superficial layer cell disappeared partly, at migration layer fined cell cluster phenomenon, the reflecting layer cell align like pillar form, the tide marks line was integrity;In low doses group cartilage surface had leakage, superficial layer cell disappear partly;at migrate layer fined cell cluster phenomenon, reflecting layer cell align like pillar form, the tide mark was discontinuation. ?Electron microscope observationIn normal group cartilage cell located in cartilage lacuna, cell wascircular with microvilli outstanding on surface, nuclear appearance was normal, nuclear membrane swollen partly, rough endoplasmic reticulum and mitochondria was observed inside cell, collagen fibers aligned regularly , periodicity transverse striation was observed on collagen fibers, different size matrix granule was observed between collagen fibers.In control group cartilage cellmorphous was oval with microvilli outstanding on surface, two nuclear membrane layer was clear , many rough endoplasmic reticulum mitochondria and collagen fibers was observed inside cell. Periodicity transverse striation was observed on collagen fibers, many matrix grain existed in collagn fiber.In model group cartilage cell appearance was irregular. Observed microtriche on surface, nuclear size decreased, cell membrane and nuclear membrane was integrity, cell organ was unclear, lipid droplet was observed in endochylema, collagen fibers align disord, diameter of collagen fibers increased.In high dose group cartilage cell morphous and microvilli was similar to control group, the expansion of RER relieve obviously, diameter of collagen fibers was small than the control group, many matrix grain was observed in the collagen fiber.In medium dose group super microstructural morphous of cartilage cell was better obviously than model group, microstructura morphous of cell organ was like the high dose, fiber morphous was better than high dose group and diameter of fiber was similar to high dose group . 1. 4 ConclusionTongLuShengGu Capsule can treat and prevent OA efficiently , eliminate imflamation reaction in rabbits joint, prevent cartilage breakdown, maintain normal morphous and function of chondrocyte, stimulate synthesis of chondcyte matrix and prevented chondrocyte degradation. 1.5 DiscussIn this experiment caroid-induced osteoarthritis animal model was used which mainly show syndrome of cartilage breakdown, chondcyte matrix degradation, swollen, synovial fluid increasing and so on .these syndrome consistent with clinical syndrome of OA. Basing on this experiment and relation research result,we infered it' s treated mechanism was relieveing high intraosseous pressure, promoting joint local blood circulation and metabolitesexcretion, maintaining articulue environmental balance and cartilage morphous and function, stimulating synthesis of chondcyte matrix, keeping banlance of chondcyte synthesis and degradation and blocking cartilage breaktion caused by inflammation mediator.2 Clinical research of TongLuShengGu Capsule Combined with Principle of Biomechanics on treating osteoarthritis.2.1 Object To evaluate the clinical effects of TongLuShengGu Capsule combined with principle of biomechanics on treating osteoarthritis.2.2 Methods2.2.1 diagnostic standard2.2.1.1 diagnostic standard of western medicineaccording to diagnose xtandard of hip and knee joint OA maded by American Rheumatism Association. 2. 2. 1. 2 diagnostic standard of Chinese medicineaccording to the diagnose standard of Syndrome of deficiency of kidney and liver, obstruction of tendonous and collaterals of OA in clinical research guidance for new drug of Chinese medicinal materials issued by Ministry of Health of the People's Republic of China. 2. 2. 3 methods of treatmentTreated group: administered with TongLuShengGu Capsule, 4 pill every time, thrid time dirly, meanwhile according to biomechanics principle Bearing Weight Under Abduce Therapy and skin traction was used.Control group: administered with Celebrex , one pill every time, once daily.2.3 ResultIn two group complaint of arthrocele and pain was relieved markly , affected limb function was improved too, satisfaction rate of treat group was 57.7%, total effective rate was 87. 3% and control group was 65. 4% and 92. 3% respectively , total treated effect of two group had no significant difference .2.4 discussPossible treated mechanism of TongLuShengGu Capsule combined with biomechanics principle as follow?Improved microcirculation of local tissue and bone, depressed intraosseous pressure, alter pathology change of Ultrastructural bone marrow, block infernal circle of high intraosseous pressure, recoverymicrocirculati, internal environment and intraosseous pressure;(2) Maintenanced normal function and morphous of chondrocyte, promoted metabolism of arthrodial cartilage, recoveried function of impaired chondrocyte, promoted synthesis of cell matrix;?Decreased secretion of MMP, promoted production of TIMP, improved index of MMP/TIMP, prevented abnormal degradation of cell matrix, prevented cartilage breakdown;?Promoted synthesis factor production, decreased destruction factor production, stimulated inflamation resorption , prevented cartilage destroy by inflammation reaction;?Applied biomechanics principle to intervent abnormal architecture and function of affected joint, increased article stability, corrected article line of force, prenvented malstress destroy cartilage, profiled cartilage repare, prevent cartilage cataplasia. 2.5 conclusionTongLuShengGu capsule conbined biomechanics principle can improved complaint of osteoarthritis, retardated osteoarthritis progression, and prevent osteoarthrits recurrence. Was a effective method for rheumatism involving bone with Syndrome of deficiency of kidney and liver, obstruction of tendonous and collaterals.