Combination Of Transcatherter Arterial Embolization And Molecular Targeted Agent For Management Of HCC: An Experimental Study

OBJECTIVE:(1)To assess the safety, feasibility and efficacy of combination transcatheter arterial embolization wih lipiodol and sorafenib.(2) To determine the slow-release effect of DC-Bead loaded-sorafenib and its impact on the normal liver of dogs. MATERIALS AND METHODSPARTⅠ.(1)In vitro study:The emulsions of sorafenid-lipiodol and cisplatin-lipiodol were prepared by mixing the drug (4mg),lipiodol (1 ml) and normal sodium. After placement for 20 minutes, the stability of sorafenid and cisplatin in lipiodol were observed..(2)In vivo study:24 New Zealand rabbits were randomly divided into three groups:groupⅠ(Lipiodol-sorafenid), groupⅡ(Lipiodol) and group III (Sorafenib). groupⅠandⅡwere treated by transcatheter selective hepatic arterial embolization with emulsions of lipiodol and sorafenib or with only lipiodol,while groupⅢwas given hepatic arterial infusion with sorafenib. Sorafenib concentration in plasma was determined by HPLC (high performance liquid chromatography) in 0 min,20min,1h,2h,4h,8h,16h,32h and 48h respectively.The breathing rate,heart rate,rectal temperature and body weight were measured,as well the.blood routine test and the function of liver, kidney,and heart.Two animals of each group were respectively killed in the 3rd day,1st,3rd and 6th week after treatment.Histopathologic study was done to liver,heart,kinney,lung,brain,gallbladder and intestine.PARTⅡ.(1) To obtain the maximal drug-loading of DC-Bead, different sizes of DC-Bead (300-500μm and 500-700μm) were tried. five bottles of different sizes of DC-Bead were added into 75% salution of sorafenid-alcohol with different concentrations:Bottle a,50mg/20ml; Bottle b, 100mg/20ml; Bottle c,100mg/40ml; Bottle d,200mg/40m; Bottle e,250mg/50ml.(2)In vivo study:16 dogs were randomly divided into four groups [group A, Sorafenib-DC-Bead(500-700nm); group B, Sorafenib-DC-Bead (300-500μm); group C, DC-Bead(300-500μm); group D, Lipiodol-sorafenib] and four dogs in each group.Each group was treated with TAE with emulsion mentioned above. Sorafenib plasma concentration was determined with HPLC in 0 min,30min,lh,4h,16h,ld,2d,4d,7d and 10 day respectively.The function of liver and kidney were observed.Experimental animals were killed and were subjected to a comprehensive necropsy that involved detailed examination of the external surface of the body and all orifices as well as the cranial,thoracic, and abdominal cavities and contents. Particular attention was paid to the liver and associated tissue and vasculature, lungs, gallbladder and bile ducts, and heart.Tissues were embedded,sectioned, and stained with hematoxylin and eosin for histopathologic assessment..RESULT:PART I:(1)In vitro study:Sorafenib was not dissolved into water or lipiodol.Compared with DDP, the emulsion of sorafenib and lipiodol was more stable in water.(2) In vivo study:①The peak sorafenib concentration (Cmax)and AUC(Area under curve) in plasma in group I was 2.46±0.101μg/ml and 945.72±52.3μg/mL.min respectively,while in group III which was 3.78±0.180 ug/ml and 546.98±21.1μg/mL.min. Compared with group III,the Cmax and AUC of group I had a significant statistics difference(p<0.05).②The breathing rate,heart rate,rectal temperature and AST/ALT,WBC,NEU% of groupⅠand groupⅢhas a significant statistics difference(p<0.05) in the 3rd day.③CK,CK-MB,DB,Cr, BUN,RBC,PLT in plasma did not change in all group.④Local necrosis was seen in group I and groupⅡin the 3rd day and 1st week,but they did not seem to be different.Group III showed no necrosis.Granulation tissue with bile dut,portal vein and micriovessels hyperplasia were seen in local necrosis area in the 3rd week.No pathological changes were found in brain,heart,kiney, intestine and gallbladder.PRATⅡ:(1) In vitro stduy:Sorafenib can be dissovled into 75% alchohol and the best concentraion for drug-loading was 100mg/20ml. (2)In vivo study:①Compared with group D,the Cmax and AUC in plasma in group A and B has a significant statistics difference(p<0.05).②Sorafenib concentarion in liver tissue could be determined in group A and B in the 3rd day and even afer one week while it could not be determined in group D.③ALT/AST in all groups increased significantly in the 3rd day after treatment.But in group B and C it changed more(P<0.05).AST/ALT returned to normal in the 1st week in group D,while in group B and C they were in a relatively high level(P<0.05).DB/ALB in all groups did not change.④The liver was injuried differently in all groups in the 3rd day and 1st week.To the tetail,group B>goup C>group A>group D.Smaller size of DC-Bead would cause more severe injury.⑤Granulation tissue with bile dut,portal vein and micriovessels hyperplasia were seen in local necrosis area.⑥One dog in group B suffered from bile duct injury.Conclusion:(1)TAE with emulsions of lipiodol and sorafenib is feasible,safe and has some slow-release effect.It seems to be more stable in water than DDP when mixed with lipiodol.(2) Sorafenib can be loaded in DC-Bead in a certain condition, such as 75% alcohol.Compared with emulsion with sorafenib and lipiodol,DC-Bead has a definite sow-release function and it is superior to lipiodol.(3)TAE with DC-Bead loaded with sorafenib is safe and feasible,but it may have an effect on the recover of injured liver.(4)Smaller particle size of DC-Bead with sorafenib may cause bile duct injury.