The Experiment Studies Of Interventional Therapy With Infusion Of Nano-hydroxyapatite Via Liver Artery On Rabbit VX2 Hepatic Tumor

PartⅠExperimental study of the effects of hydroxyapatitenanoparticle at different dosages on liverVX2 tumor growth in rabbitsObjective To study the effects of hydroxyapatite nanoparticle(nHAP) at different dosageson liver tumor growth in rabbits.Methods Sixty New Zealand white rabbits were randomly divided into four groups, eachhaving 15 rabbits, and VX2 carcinoma was implanted in the left lobes of the liver. Twoweeks later, a catheter was inserted into the gastroduenal artery of the rabbit and nHAP atdifferent dosages were injected into rabbits via hepatic artery. Group A(Control group wasinjected with 5 ml saline); Group B(Low-nHAP group with 0.5mg.kg^-1); Group C(Middle-nHAP group with 5mg.kg^-1); Group D(High-nHAP group with 50mg.kg^-1); SpiralCT scan was performed to measure tumor volume 7 days and 14 days after treatment. Theserum levels of AST and ALT were observed 1 day and 7 days after treatment. The survivalsof VX2 rabbits after treatment were also observed.Results The tumor growth rates of group C and D in 7 days and 14 days after treatment were lower than those in control group(p＜0.05), but they were not lower in group B thanthat in control group. The life span of group C or group D was longer than that in controlgroup(p＜0.05), it is not longer in group B than that in control group(p＞0.05). The serumlevel of AST or ALT in group D was higher than that in the other three groups 1 day aftertreatment(p＜0.05), but the serum level of AST or ALT was normal in all groups 7 days aftertreatment.Conclusion The results showed that nHAP at 5-50mg.kg^-1 has signifigant inhibitoryeffect on liver VX2 tumor growth in rabbits with minimal liver toxicity, and prolongs thelife span of the rabbits with VX2 liver tumors.PartⅡThe effects of lipiodol-hydroxyapatite nanoparticle viahepatic arterial infusion on tumor's growth and angiogenesis inrabbit VX2 hepatic tumor cellObjective To investigate the effects of lipiodol-hydroxyapatite nanoparticle(lipi-nHAP)via the hepatic arterial infusion on tumor's growth, necrotic rate and angiogenesis inrabbit VX2 hepatic tumor.Methods One hundred New Zealand white rabbits were randomly divided into fivegroups, each group having 20 rabbits, and VX2 carcinoma was implanted in the left lobesof the liver. Two weeks later, abdominal exposure was carried out through an upper midlineabdominal incision, a catheter was inserted into the gastroduenal artery of the rabbit withVX2 hepatic tumor and infusion was performed via the hepatic artery using physiologicalsaline(group A), nHAP(group B), Lipiodol(group C), ADM adding Lipiodol(group D)and Lipi-nHAP(group E). The ALT and AST preoperation and postoperation wereexamined. The size of tumors was observed by Spiral CT Scan, the volume and growthrates of tumors were calculated, necrotic rates of tumors were assessed using pathologicalslices way. The values of MVD and the expression level of VEGF in survival tumor tissuewere examined two weeks after treatment by immunohistochemistry with anti-CD31,anti-VEGF monoclonal antibody respectively. The survivals of the rabbits after treatment were also recorded.Results There is no significant difference between the groups for the ALT and ASTpreoperation and postoperation. The tumor volumes and growth rates of group E in 7 daysand 14 days after treatment were lower than those in other groups(P＜0.05). The necroticrates in group E were higher than those in other groups(P＜0.05). The values of MVD andthe expression level VEGF were higher in groups C and D compared with those in group A.Compared with those in other groups, The values of MVD and the expression level ofVEGF were significantly lower in group E(P＜0.05). The survivals of group E werelonger than those in other groups(P＜0.05).Conclusion Lipi-nHAP can suppress the growth of tumors, increase the tumor's necroticrates and inhibit the development of neovascularization, decrease the expression level ofVEGF of residual tumors, prolong the survivals of the rabbits with VX2 hepatic tumorsand also safe to the liver function.PartⅢThe effects of 5-Fluorouracil-lipiodol-hydroxyapatitenanoparticle via hepatic arterial infusion on tumor's growth, apoptosis,proliferation and angiogenesis in rabbit VX2 hepatic tumor cellObjective To investigate the effects of 5-Fluorouracil-lipiodol-hydroxyapatitenanoparticle(5-Fu-lipi-nHAP) via the hepatic arterial infusion on tumor's growth,necrotic rate, apoptosis, proliferation and angiogenesis in rabbit VX2 hepatic tumor.Methods One hundred New Zealand white rabbits were randomly divided into fivegroups, each group having 20 rabbits, and VX2 carcinoma was implanted in the left lobesof the liver. Two weeks later, abdominal exposure was carried out through an upper midlineabdominal incision, a catheter was inserted into the gastroduenal artery of the rabbit withVX2 hepatic tumor and infusion was performed via the hepatic artery using physiologicalsaline(group A), Lipiodol(group B), 5-Fu adding Lipiodol(group C), Lipi-nHAP (groupD) and 5-Fu-lipi-nHAP(E). The size of tumors was observed by Spiral CT Scan, the volume and growth rates of tumors were calculated, necrotic rates of tumors were assessedusing pathological slices way. The apoptotic index, proliferation index and values of MVDand VEGF in survival tumor tissue were examined two weeks after treatment by TUNELmethod and immunohistochemistry with anti-CD31, anti-VEGF, anti-PCNA monoclonalantibody respectively. The survivals of the rabbits after treatment were also recorded.Results The tumor volumes and growth rates of group E in 7 days and 14 days aftertreatment were lower than those in other groups(P＜0.05). The necrotic rates in group Ewere higher than those in other groups(P＜0.05). The apoptotic index in group D and ingroup E were higher than those in other groups(P＜0.05). The values of MVD and VEGFwere higher in groups C and D compared with those in group A. Compared with those inother groups, The values of MVD, the values of VEGF and expression level of PCNA weresignificantly lower in group D and in group E(P＜0.05). The survivals of group E werelonger than those in other groups(P＜0.05).Conclusion 5-Fu-Lipi-nHAP can suppress the growth of tumors, increase the tumor'snecrotic rates and apoptotic index, inhibit the development of neovascularization, decreasethe expression level of PCNA of residual tumors and prolong the survivals of the rabbitswith VX2 hepatic tumors.PartⅣPreparation of lipiodol-nanohydroxyapatie and it's targetingin vx2 tumor bearing rabbits while infusion via hepatic arteryObjective To study method of preparing lipido-nano hydroxyapatite by ultrasonicemulsification and observe it's targeting in vx2 tumor beating rabbits with TransmissionElectron Microscope after it's infusion via hepatic artery.Methods Mix iodinated oil and nano hydroxyapatite at the propotion of 3ml:50mg, thenemulsify the mixture with ultrasound, observe it's appearance under Transmission ElectronMicroscope. Making 24 tumor bearing rabbits models, and randomly dividing the 24rabbits into 3 groups two weeks later, with each group 8 rabbits. Infuse the mixture via hepatic artery in each group, and observe it's distribution in liver, spleen, brain, kidney andheart under tasnsmission electron microscope at 1h, 24h and 48h after infusion.Results The lipido-nano hydroxyapatite, made with the ultrasonic emulfication method, hasa good uniformity, and it's diameter is between 10～100nm. We can observe the particles inliver and tumor cells lh after infusion and only in tumor cells 24h later. Conclusion: Tthemethod is simple and easy to make lipido-nano hydroxyapatite. The particles are mainlydistributed in liver and tumor cells after it's infusion via hepatic artery, and has a goodtargrting at liver tumor.