Application Of Home-made Puncture Instrument For Diagnosis Of Bone Tumor In Animal Models

Objective:To investigate the application of the home-made puncture instrument for experimental rabbit to control puncturing complication after establishing rabbit model with VX2 bone tumor.Methods:Forty-three New Zealand male rabbits were purchased from national rabbit farm.The VX2 tumor cell were purchased from reagents corporation.1、The VX2 muscle tumor model with transplanted by using tumor cells suspension injection which was selected randomly was successfully established. At 2 weeks after implanted,it was confirmed that VX2 tumor goitre existed in rabbit’s thigh muscle by generally image and Color Doppler Ultra Sound examination.In this way,we got a large number of VX2 tumor cells.A part of them was used in subsequent experiment.The rest of them was cryo-preserved in nitrogen canister.The way of planting tumor tissue suspension was applied for establishing the rabbit model with VX2 bone tumor.2、Ten experimental rabbits were randomly selected and were established into bone tumor models by using tumor cells suspension injection.We executed two models in every two weeks.Growth of bone tumor cells were observed by generally image 、 X-ray examination and pathological examination.By these means,we could assess the time(easy to puncture、obtain enough tissue samples and block off puncture hole)of contralateral bone cortex bursting.3、The left tibia of thirty rabbit models were planted VX2 tumor cells.The right same position of thirty rabbit models were untreated.Thirty rabbit models were randomly divided in the experimental(n=15) and control groups(n=15).In the experimental group,puncture biopsy was performed by a home-made puncture instrument and the bone holes were occluded subsequently.In the control group,puncture biopsy was conducted by using a conventional punctue instrument whereas the holes were left open..At 1h after puncture,we used Color Doppler Ultra Sound to evaluate the bleeding condition of the right bone holes.At 2 weeks after puncture,all the rabbits were executed,evaluated and compared the tumor progression on histopathological analysis.Statistical process was used by SPSS 19.0 of statistical software. P<0.01 was considered statistically different between of them.Results:1、We successfully establish rabbit VX2 muscle tumor model and got a large number of VX2 tumor tissues.2、Ten rabbits which were used to establish VX2 bone tumor models were put to death regularly in batches.The VX2 tumor cells developed in tibial marrow in 1 or 2 weeks.It developed in marrow and broke through bone cortex in 2 or 3 weeks.It infiltrated surrounding soft tissue in 3 or 4 weeks.It confirmed that the suitable time for using home-made puncture instrument on aspiration biopsy is in the third week.3、Thirty rabbits were used to establish VX2 bone tumor models.(1)At 1h after puncture,color ultrasound revealed no signs of device separation.Hemorrhage and hematoma were observed around the puncture hole in 2 animals(13.33%) and no apparent fluid effusion was detected in the remaining 13 in the experimental group,significantly different from those in the control group,14 animals(93.33%) had hemorrhage and hematoma surrounding the puncture hole and the other 1 had no evident hemorrhage(P<0.01).The color ultrasound of right tibia soft tissue showed that our home-made puncture instrument and technology were superior to traditional puncture device and technology in controlling bleeding statistically.(2)At 2 weeks after puncture,histopathological observation revealed no signs of device separation.No apparent tumor spread was detected in the experimental group,significantly different from those in the control group,14 animals(93.33%) had tumor spread(P<0.01). The histopathological observation of left tibia soft tissue showed that after two weeks from puncturing our home-made puncture instrument and technology were superior to traditional puncture device and technology in controlling tumor spread statistically.Conclusion:Our home-made puncture instrument and technology can be used for the study of bone tumor model.It is able to obtain sufficient pathological organization and can control the bleeding of the puncture hole by implanting the bolt through the cannula, thereby controlling the local diffusion of tumor. Since all the operations are done in the cannula,it also can reduce the pollution of the pathological tissue for puncture pipage. The cannula addes on the basis of the traditional puncture device form a channel between the tumor and outside. We obtain the tumor tissue in the sleeve,and implant the bolt to puncture pipage through the cannula,then pull out the sleeve, so we can maximize to maintain the integrity of the boundary between the bone tumor and normal tissue,thus prevent bleeding and tumor spread.Our experiments show that this home-made puncture instrument and technology is simple, safe and available with less complication of the puncture pipage.It shows a great clinical application value in the subsequent bone tumor study.