Acupuncture For Osteoporostic Fracture-A Clinical And Experimental Study

Osteoporotic fracture, so-called brittle fracture, which is caused byminor violence or non-traumatic factors, is due to low bone mineral density(BMD), abnormality of bony microstructure, enhancement of thebone-fragility going along with aging. As a most serious complication ofosteoporosis, osteoporotic fracture badly harm the health of elderly people,so extensive attention from all society should be paid to it .1 Clinical study: The short-termacesodyne effects of acupuncture on earlyosteoporostic compression fracture of thoracic and lumber vertebrae1.1 Tentative plan and the aim of studyAccording to previous studies and literatures, acupuncture and moxibustionfor osteoporosiscan increases the BMD, ameliorate clinical symptoms, improvebony metabolism, and enhance the biodynatic capability of skeletons. Also,Acupuncture and moxibustions can promote union of fracture, and relievepain. can promote union of fracture, and relieve pain.As the most common complication of osteoporosis, osteoporosticcompression fracture of thoracic and lumber vertebrae (OCFTLV), whichmanifest severe low back pain. dysfunction of waist and deformity ofspine, etc, badly harm the health of elderly people, threatens health andaffects living quality. The pain in the early stage of the fracture canimmediately disturb the patients to taking active exercise in early stage,lead to ill-timed reduction and result poor outcomes. So, it is ofsignificance to relieve the pain timelyIn order evaluate the acesodyne effect of acupuncture and moxibustion onthe early stage of fresh throacidc and lumber vertebrae, we designed thisrandom control clinical study1.2 Patients and methods sixty-four patients with OCFTLV were randomly allocated to two groups:Group A, thirty-four cases were acupunctured electrically daily onbilateral Weizhong Points and bilateral Huatuo Jiaji Points adjacent to theinjured vertebrae. Group B, thirty patients were administrated with Calcininand calcium carbonate 115g, vitamin D 125IU, (1 tablet, po, qd). The periodof treatment was two-week. Patients-Self-VAS Scoring and Doctor-Check-TenderScoring were applied to evaluate the ache before and after the treatments.1.3 ResultsAfter two-week treatment, statistically significant improvement inPatients-Self-YAS Scoring(including spontaneous low back pain, moving painand turning-over pain) and Doctor-Check-Tender Scoring were detected(P＜0.01). No serious adverse event occurred in the two groups.1.4 ConclusionsThe short-term acesodyne effect of acupuncture for low back pain inpatients with OCFTLV was tangible. which was similar with that of thecombination of calcitonin and calcium carbonate. Besides, the acupuncturetherapy deserves being generalized because it is cheaper.2 Experimental Study: The effects of acupuncture on neurotrophic factorsand their receptors in fracture callus in ovariecomized rats2.1 Tentative plan and the aim of studyThe previous studies on Expression of neurotrophic factors (NTFs)and theirreceptors during fracture healing showed that several kinds of NTFs,including nerve growth factor (NGF), brain-derived neurotrophic factor(BDNF) and neurotrophin-3 (NT-3)are expressed during fracture healing; while TrkA, TrkB and TrkC, thereceptors of NTFs, are also expressed in osteoblast-like cells .Thesephenomena suggest that NTFs may be involved in bone formation, but themechanism is unclear. Postmenopausal osteoporosis(PMO) is closely relatedto the postmenopausal decrease of estrogen. Moreover, it has been found thatthere are many physiological and pharmacological resemblances,correlationsand superposition between estrogen and BDNF in CNS. However, the reciprocityand relation between estrogen and BDNF during fracture healing have yet notbeen reported.In order to clarify cellularly and molecularly the roles of estrogen,neurotrophins and their receptors(TrkA, TrkB and TrkC) in bone formation during osteoporotic fracture healing, based on the TCM theories of "theStandpoint of Integrity", "Kidney controlling bone" and "Marrowprocreating bone", we make animal model of Typeâ… osteoporostic fractureand applied immunostaining technique to investigate the localizations of nervegrowth factor (NGF), brain-derived neurotrophic factor (BDNF), andneurotrophin-3 (NT-3) and their receptors at the fracture callus. Probingfurther into the pathological mechanism may provide new therapeutic strategyfor osteoporostic fracture. Also, from this study, we may know more aboutthe effects of acupuncture on NTFs in the callus of ovariecomized female rats,which may provide new paths for preventing and treating osteoporosis2.2 Materials and methods2.2.1 Grouping the animals Sixty healthy female SD rats, 3-monthold, non-pregnant, weighting 300±20g, were randomly allocated to four groups:normal group(Group A), model group(Group B), acupuncture group (Group C) andnilestriol group(Group D). To make model of postmenopausal osteoporosis(PMOP),in all groups but Group A, the rats were overiectomized, while in Group A,for sham operation as a control, ovaries of 15 rats was similarly exposedbut not resected .2.2.2 Serology examination Three days before ovariectomy and beforesacrifice, all rats were phlebotomized to test the levels of E₂ in serum.2.2.3 Making fracture models Three months after the ovariectomy, we tooktwo rats respectively from each group to scanned generally with DEXA. Theresults of DEXA showed that the BMD levels in Group B,C,D were much lowerthan those in GroupA, which meant that the rats in Group B,C,D had sufferedPMOP. Then, referring to literatures, under anaesthesia of Chlorali Hydras,the left femurs of all rats were closely fractured and fixed intramedully with1mm Kirschner wire. To prevent infection, after the operation, every ratswere injected intramuscularly with PenicillinumG, 40000 U, qd, for threedays. The animals were provided with standard feedstuff and running waterwithout limitation.2.2.4 Alteration of weights All rats were weighed weekly to observe thechanges of rats, weights in each groups.2.2.5 Treatment of fracture Four days after the let femurs were fractured,treatment began.The rats in the Group A (normal fracture control)and Group B (PMO fracture control) were given orally normal saline solution, 3ml, weekly.Group C (PMO fracture treated with acupuncture) were treated withacupunctureAcupuncture points: Huantioa, Housanli, Yanglingchuan and Weizhong onthe left hind legs (fractured)Acupuncture methods: On each rats in Group C, after aiming correctlyat the points, punctured vertically with sterilized 25mm milli-needles,twist each needle for 25 seconds every other 5 minutes. The needles were leftfor 20 minutes. The therapy was practiced daily.Group D (PMO fracture treated with nilestriol): The rats in the Group Dwere given orally nilestriol solution in a doses of 0.6. ml/100g weight(concentration of nilestriol: 0.2mg/ml), 3ml, weekly.2.2.6 Harvesting samples After starting the treatment, two or three ratsfrom each group were randomly selected to be executed for harvestingsamples. Before decollation, each animal was weighed and phlebotomized to testthe levels of E₂, in serum.The callus of left hind femur fracture were taken and each samples weredivided into 2 parts: one for histological examinations, and other one forimmunostaining examination of NTFS and Trks.2.2.7 The observing item2.2.7.1 Histological examination: the callus sections were stained with HEstain.2.2. 7. 2 Immunostaining examination (SABC method): Polyclonal antibodiesagainst TRKA, TRKB, TRKC, NGF, BDNF, and NT-3 were dripped to the callussections, then, reaction was visualized by immersing sections indimethylbenzidine, and restained with campeachy. Positive colour was brown.The expression level of NTFs and Trks were measured quantitively withHPIAS-100 High Picture Image Analysis System-100 (HPIAS-100). Three viewswere randomly chosen in each slides to measure the grey gradients of NTFs andTrks, which were then disposed automatically by computer to produce theaverage value of grey gradients of each slide.2.2.7.3 The test of BMD of isolated right femur After the rats weresacrificed, right femur of each rat was taken out and eliminated the musclesattached. Then the BMD of the isolated right femurs were tested withDEXA(QDR-4000, Hologic) 2.3 Results2.3.1 WeightsOne months after the ovariectomy, the weights of Group B, C and D increasedgreatly, statistically significantly different from those of themselvesbefore the surgery.. Just before execution, the differences between weightsof B, C, D and that of A Group, which was lighter, were of statisticalsignificance .2.3.2 The levels of E₂ in serumBefore the ovariectomy, the differences of the levels of E₂ in serumbetweenevery two groups were not statistically significantly different. Just beforeexecution, the differences between E₂ levels of B, C, D Group and that ofA Group, which was higher, were of statistical significance. ComparingE₂ levels of B, C Group with that of D Group, E₂ level was relatively higherin D Group with statistical significance.2.3.3. BMD Three months after the ovariectomy, comparing to the generalBMD of A Group, those of B, C, D Group were obvious lower with statisticalsignificance. While comparison of the BMD of the isolated right femurs amongall groups showed no statistical significance.2.3.4 Optical observation of HE stained slides HE stained callus slidesobserved by optical microscope showed that the process of fracture healingin A,C,D Group was faster than that in Group B.2.3.5 Immunostaining Histological observation Positive Immunostaining ofNGF, BDNF, NT-3, TrkA, TrkB and TrkC could be seen in every group, mainly in7 and 14days after the fractures. The locations of the expression includedmainly bone -forming cells such as osteoblast, osteoblast-like cells,osteocyte-like cells,proliferating chondrocytes, mature chondrocytes, andhypertrophic chondrocytes, as well as a few in macrophages multinucleate-macrophages in marrow tissue. The expression levels from high to low in turnwere A, C , D then BGroup..The "average grey gradients" comparison of A and B Group, and also, B andC, D Group showed statistical significant difference.2.4 Conclusion2.4.1 In this study, rats' models of osteoporostic fracture was made byovariectomy plus close fracturing..This method is believed to be correctlylocating, conveniently practicing,credible and minor traumatic. The severities of injury and internal fixation are similar and comparable in everygroup. In addition, the pathological alterations are similar to those ofclinic. It is an ideal animal model for study of osteoporostic fracture.2.4.2 Expressions of NGF, BDNF, NT-3, TrkA, TrkB and TrkC occur both incalluses of normal fracture and osteoporostic fracture, but the expressionswere less in osteoporostic fracture.2.4.3 The levels of E₂ in serum can influenced the expressions of NGF, BDNF,NT-3, TrkA, TrkB and TrkC in callus; decrease of E₂ level can lead to decreaseof.expressions of NTFs and their receptors in callus. Supplementing estrogencan raise the levels of E₂ in serum as well as the expression of NTFs and theirreceptors in callus.2.4.4 Although acupuncture can not elevate the levels of E₂ in serum, it canpartly mitigate the decrease of expression of NTFs and their receptors incallus caused by ovariectomy in some degree.