Experimental Research Of The DieChuang San Combined With Transdermal Drug To Promote Healing Of The Closed Fracture

ObjectiveBy observing changes of the concentration for ALP,Ca²⁺,P³⁺in DiechuangSan （DCS） combined with the transdermal drug to promote rabbits closedfracture healing process,the changes of X-ray different periods,the callushistopathology and callus change in the molecular level the expression ofVEGF, to study the affect of the DCS combined with the topical agents inclosed fracture healing.MethodsRabbit age about six months, male New Zealand white rabbits60. toadaptive feeding after7days, use of Weight1250g, the thickness of1.0cmtraction mound downward impact, create the rabbit left tibia closed transversefracture in the lower segmen（Because the rabbit fibula and tibia in the lowersegment fusion）, manual reduction, retrograde closed into the Kirschner wireto the tibia at the lateral malleolus of the tibia in the aseptic, result in lowersegment of the left tibia closed fracture model, after rabbits successful models,randomly divided into group:DSC combined with3%menthol treatment group,DCS group, Yunnanbaiyao group and the blank control group （n=15）, fracturefor given different drugs for topical dressing was changed every3days,batches2,4and8weeks after administration, respectively （n=5） take about2mL of blood from the carotid artery after anesthesia. The measured the ALP,Ca²⁺, P³⁺concentrations, and after the the bloodletting death of the animals inanesthetized state, left hind leg X-ray radiography examination, observation of the closed fracture healing and callus growth, after radiography, at the centerof the left tibia bone defect, take a length of about1cm of the bone segment,placing it fixed with4%formaldehyde solution4°C24hours. Then be placedin the mass fraction of15%EDTA-2Na solution4℃decalcification, low meltingpoint paraffin-embedded, cut into the thickness of about5μm, were stained with HE. To observed thenumber of callus capillaries and osteogenic cells, immunohistochemicalstaining of callus of vascular endothelial growth factor （VEGF） expression.Results1．the serum biochemical tests resultsThe DCS combined with the transdermal drug can significantly improvethe experimental animals at different times of serum ALP and theconcentration of P³⁺, and the peak early appearance, compared with the otherthree groups was statistically significant （P <0.05）.the Ca²⁺concentration earlyrise, the latter decreased, so early appearance of the peak of the Ca·P product,promote calcium deposition, and accelerated fracture healing.2．X-ray Analysis and the Lane’s score systemThe DCS combined with the transdermal drug exogenous osteotylusappeared early, quantity and the canal recanalization appear earlier than othergroups, there is significant difference of the Lane’s scores,compared with theother three groups （P <0.05）, the purely DCS group and the Yunnanbaiyaogroup callus growth rate is almost the same, less callus in the entireobservation period, no significant canal recanalization. the blank control groupbe seen a small amount of callus in the latter, fracture line is still clearly visible.3．Tissue sections by HE staining resultsThe DCS combined with the transdermal drug can be seen a largenumber of chondrocytes and osteoblasts early, and more capillaries, and astime goes on, the trabecular bone and capillaries arranged in neat rows earlier,the purely DCS group and Yunnanbaiyao group the cartilage cells and osteoblasts are relatively small in the closed fracture healing process, andoccasionally the capillary distribution, the blank control group seen only a smallamount of blood vessels in the latter, and the distribution is disordered.4．The results of immunohistochemistryThe DCS combined with the transdermal drug VEGF levels in the callustissue is very abundant and strongly positive expression, the purely DCS groupand Yunnanbaiyao group have positive expression in the callus in the sameperiod, but its expression level is weak. the blank control group expression ofvery few, between the four groups is very significant differences.ConclusionsThe DCS combined with the transdermal drug topical significantlypromote calcium deposition in the fracture site, improve the blood circulation ofin the fracture, significantly promote callus capillaries、 osteoblasts andcartilage cells, the early formation of the trabecular bone, haversian system inadvance to mature, thus speeding up the fracture healing, shorten the healingtime.