Experimental Investigation On Chinese Medicine Pyritum With Low Frequency Ultrasound Treatment For Heal Fracture

Background:Pyritum is important Chinese medicine for bone fracture and recorded in all ages in our national pharmacopoeia codex.It can help improving blood circulation and relieving bruises and pains and muscle and ligament reunion of bone.It is clinically used for injuries from falls,muscle and bone broken,gore and pains etc.Pyritum is the most common used medicine for bone-knitting and symplectic bone which there is no other orthopaedics prescription to replace.Pyritum is always used matching with other Chinese medicine clinically and in multi-oral prescription.A few of them are used in marinate and electuary ointment.Bone broken always occurs locally.As Pyritum has such an effection in treatment of fracture,it must he a ideal method to administrate locally with Pyritum which can induce a high local drug concentration and drug plays a role locally that there is little drug concentration in body plasma.Hyodermic is one method of it.However it is painful,multi-point needed,difficult to master the level,not well-distrbuted.The disadvantage is low drug concentration in less plasma position.Through using a certain level low frequency ultrasound, giant molecule such as protein can be obtained across the skin arriving on target which will not be affected by local blood circle.It can realize the acquisition of local body insufficient of plasma and drug well-distribute.So local small dose across skin can make a relatively high concentration and has an important effects;meanwhile it has a sustained release effective.There is a little dosage in plasma and it is safe.Also it reduces secondary reaction.Object:After administrating Chinese medicine Pyritum with low frequency ultrasound,we observed local effective agent concentration and the effects of BMp-2's expression on fracture restitution when using Pyritum and low frequency ultrasound's administration effects on rabbit's bone restitution.Method:1.Put 75 New Zealand white big ears rabbits into 5 groups randomly,15 each.Method:one treatment group:0.5g/ml Pyritum decoction;two treatment groups:1.0g/ml Pyritum decoction;three treatment groups: 2.0g/ml Pyritum decoction.Group 1-3 with low frequency ultrasound, once per day.Control group:ultrasound with frequency of 20KHz, intensity of 100mw/cm²,low frequency in vitro irradiation,once per day,20min once.Model group:normal saline 20ml lavage,once per day.2.Pyritum preparation:Pyritum pick up impurity strictly,smash the ore, heat it in fire and dip it into vinegar:heat it to dark red,dip it into vinegar to pitchy,luster disappears and loose(if not loose over and over again till loose )vinegar 30kg per 100kg Pyritum(NF. Pharmacopoeia of PRC2005 edition(the lst):95)3.X-ray transmission4.poroma dry5.determination of Fe,Cu,Mn,Zn in poroma6.hybrid7.develop a film8.determination of Ca²⁺,P ion concentration and Alp activity in plasma 9.statistics resultsResultsPyritum can obviously increase Fe,Cu contents in poroma of rabbits.(P＜0.05),and then increase poroma's growth.We can observe that poroma in experimental group is more than it in control group when using X-ray.Observed from 4 trace elements from the testing,among 3 Pyritum experimental groups Fe concentration reaches 1186μg/ml,while it reaches(172.68±14.32μg/ml)in rabbit poroma.Cu in poroma in experimental group is higher than it in control group.All Pyritum experimental groups are b(?)n,BMP-2 to express masculine. There is no color development in all control groups and model groups. All samples outside periosteum of model and control group grew thicker. Most samples of fracture mechanocyte,nerve structure and osteoid increased and fiber arrayed regularly in experimental group 1,2.All samples outside periosteum osteoplast got hyperplasia.The hyperplasia got together,bone matrix adipopexised and informed osteoid tissue,cartilage and bone trabecula,and expanded towards fracture clearance.Lame cartilage and bone trabecula's poroma connected and expressed masculine in mechanocyte,chondroblast cartilage,osteoplast and osteocyte.In 3 experimental groups woven bone got grow and medullary space bone trabecula got thinner and medullary space got open.It improved Ca,P,ALP concentration that union got increased.Pyritum can increase microelement in rabbit poroma obviously(p＜0.05),increase poroma growth and then improve bone repairing.Conclusion:1.Low frequency(20KHz)ultrasound administration is a safe,effective local percutaneous administration method.2.With low frequency ultrasound,heated Pyritum can be administrated across skin and has bioeffects on local body. 3.With low frequency ultrasound local concentration will increase as dosage increase and single low frequency has effects on wound healing.4.With low frequency ultrasound Pyritum can induce BMP-2 to make broken healing and improve Ca,P,ALP concentration to heal rabbit broken bone.Pyritum can increase trace element in rabbit poroma and then improve course of repair.