Akinaga Sonoda

An initial clinical study on the efficacy of cisplatin-releasing gelatin microspheres for metastatic liver tumors

PURPOSE To evaluate the antitumor effect and side effects of cisplatin-releasing gelatin microspheres (Cis-GMSs) for metastatic liver tumors. METHODS Cis-GMSs that degraded over 14 days were employed. The subjects comprised a total of nine cases. Transcatheter hepatic artery embolization (TAE) using Cis-GMSs (Cis-GMSs-TAE) was performed 13 times in total. Six cases, each containing one to five tumors in a single segment to an entire lobe were treated by Cis-GMSs-TAE. In the remaining three cases with six or more metastatic liver tumors, the right and left lobes were treated by Cis-GMSs-TAE at a 2-week interval. RESULTS There were two complete response (CR), one partial response (PR) and six stable disease (SD) cases. The response rate was 33.3%. The average rate of reduction in tumor diameter was 32%. Disappearance of metastatic liver tumors was observed in only two of the nine cases. As for side effects and complications, post-embolization syndrome was observed in eight cases, but no severe complications such as cholangitis or liver abscess were observed. CONCLUSION Considering the mild side effects by Cis-GMSs-TAE, it is suggested that Cis-GMSs-TAE should be tried at least once as topical therapy for metastatic liver tumors when the response to systemic chemotherapy and other treatments is not satisfactory.

Vasospasm of Atherosclerotic Coronary Arteries Precipitates Acute Ischemic Myocardial Damage in Myocardial Infarction–Prone Strain of the Watanabe Heritable Hyperlipidemic Rabbits

Objective—This study tested the hypothesis that vasospasm can trigger coronary plaque injury and acute ischemic myocardial damage. Approach and Results—Myocardial infarction–prone strain of the Watanabe heritable hyperlipidemic rabbits received an intravenous bolus of ergonovine maleate (0.45 µmol/kg) during intravenous infusion of norepinephrine (12 nmol/kg per minute) to provoke coronary spasm in vivo. After this treatment, coronary angiography demonstrated vasospasm, and the ECG showed ischemic abnormalities (ST depression/elevation and T-wave inversion) in 77% of animals (23/30). These changes normalized after nitroglycerin injection. In rabbits that demonstrated these ECG findings for >20 minutes, echocardiograms showed left ventricular wall motion abnormality. Serum levels of heart-type fatty acid–binding protein, cardiac troponin-I, and myoglobin increased markedly 4 hours after spasm provocation. In coronary lesions of myocardial infarction–prone strain of the Watanabe heritable hyperlipidemic rabbits with provoked coronary spasm, we observed intimal injury in 60.9% in the form of endothelial cell protrusions (39.1%), denudation (30.4%), and macrophage extravasation (56.5%). Plaque disruption with luminal thrombus, however, was only seen in 2 of 23 animals (8.7%), and mural microthrombus was rarely observed (4.3%). Conclusions—These observations show that provocation of vasospasm in myocardial infarction–prone strain of the Watanabe heritable hyperlipidemic rabbits associates with subsequent ischemic myocardial damage. Although treatment with spasmogens altered aspects of plaque morphology, for example, endothelial protrusion and macrophage emigration, thrombosis was rare in these animals with chronic atherosclerotic disease.

Atherosclerotic imaging using 4 types of superparamagnetic iron oxides: new possibilities for mannan-coated particles.

PURPOSE We used magnetic resonance imaging (MRI) and histologic techniques to compare the uptake by the rabbit atherosclerotic wall of 4 types of superparamagnetic iron oxide (SPIO) particles, i.e. SPIO, mannan-coated SPIO (M-SPIO), ultrasmall SPIO (USPIO), and mannan-coated USPIO (M-USPIO). MATERIALS AND METHODS All experimental protocols were approved by our institutional animal experimentation committee. We intravenously injected 12 Watanabe heritable hyperlipidemic rabbits with one of the 4 types of SPIO (0.8 mmol Fe/kg). Two other rabbits served as the control. The rabbits underwent in vivo contrast-enhanced magnetic resonance angiography (MRA) before- and 5 days after these injections; excised aortae were subjected to in vitro MRI. In the in vivo and in vitro studies we assessed the signal intensity of the vessels at identical regions of interest (ROI) and calculated the signal-to-noise ratio (SNR). For histologic assessment we evaluated the iron-positive regions in Prussian blue-stained specimens. RESULTS There were significant differences in iron-positive regions where M-USPIO>USPIO, M-SPIO>SPIO, USPIO>SPIO (p<0.05) but not between M-USPIO and M-SPIO. The difference between the pre- and post-injection SNR was significantly greater in rabbits treated with M-USPIO than USPIO and in rabbits injected with M-SPIO than SPIO (p<0.05). On in vitro MRI scans SNR tended to be lower in M-USPIO- and M-SPIO- than USPIO- and SPIO-treated rabbits (p<0.1). CONCLUSION Histologic and imaging analysis showed that mannan-coated SPIO and USPIO particles were taken up more readily by the atherosclerotic rabbit wall than uncoated SPIO and USPIO.

Histological study of the biodynamics of iron oxide nanoparticles with different diameters.

The biodynamics of ultrasmall and small superparamagnetic iron oxide (USPIO and SPIO, respectively) particles that were injected intraperitoneally into 36 C57BL/6 mice were investigated chronologically. Their distribution was studied histologically at six time points by measuring iron-positive areas (μm2) in organ sections stained with Prussian blue. The uptake of the differently sized particles was also compared by cultured murine macrophages (J774.1). Iron-positive areas in the liver were significantly larger in the mice injected with USPIO than those injected with SPIO at the first three time points (P < 0.05). The amount of USPIO in the lung parenchyma around the airway was larger than that of SPIO at four time points (P < 0.05); distribution to the lymph nodes was not significantly different. The amount of iron was significantly larger in SPIO- than USPIO-treated cultured cells (P < 0.05). In conclusion, it is suggested that intra peritoneally injected USPIO particles could be used more quickly than SPIO to make Kupffer images of the liver and that both agents could help get lymph node images of similar quality.

Complex comprised of dextran magnetite and conjugated cisplatin exhibiting selective hyperthermic and controlled-release potential.

We developed a dextran-magnetite conjugated cisplatin (DM-Cis) complex for use in thermal ablation and as a chemotherapeutic drug. To produce DM-Cis we reacted Cis with 1 mL DM (56 mg/mL iron). The temperature rise of DM-Cis was measured in vitro and in vivo under a portable induction-heating (IH) device. Platinum desorption from DM-Cis over 24 hours was measured in bovine serum. In in vivo accumulation and magnet and exothermic experiments we used four rat groups. In group 1 we delivered DM-Cis intraperitoneally (ip) and placed magnets subcutaneously (sc). In group 2 we injected saline (ip) and placed magnets (sc). In group 3 we injected DM-Cis (ip) and placed a sc incision (sham). The control (group 4) received an ip injection of saline. Rectus abdominis muscle tissue was stained with hematoxylin-eosin and iron-stained tissue areas (μm2) were calculated. The maximum platinum concentration in DM-Cis was approximately 105.6 μg/mL. Over 24 hours, 33.48% of platinum from DM-Cis was released. There was a significant difference (P < 0.05) in the iron-stained area between group 1 and the other groups. The temperature in muscle tissue registered a maximum of 56°C after about 4 min. DM-Cis may represent a magnetically-accumulated anticancer drug with hyperthermic effects.

Prolonged local persistence of cisplatin-loaded gelatin microspheres and their chemoembolic anti-cancer effect in rabbits

PURPOSE To confirm prolonged cisplatin release from drug-loaded gelatin microspheres (GMSs) and their improved chemoembolic anti-cancer effect against VX2 liver tumors in rabbits. MATERIALS AND METHODS Two groups of twelve rabbits each were treated intraarterially either with 2 mg/kg cisplatin-loaded GMSs (=0.04 mg/kg cisplatin) or 0.04 mg/kg cisplatin solution by administering them into the right renal artery. Platinum concentrations within the renal parenchyma were analyzed immediately following infusion (day 0) and on days 1, 3, and 7 using the atomic absorption method. In a second experiment four groups of five rabbits each with implanted VX2 liver tumors were treated intraarterially through the hepatic artery with the following drugs: 2 mg/kg cisplatin-loaded GMSs (=0.04 mg/kg cisplatin) (group I), 2 mg/kg GMSs without any drug (group II), 1.5 mg/kg cisplatin solution (group III) and saline (group IV). Tumor volumes were analyzed pre-injection and 7 days after with MRI allowing calculating the relative tumor growth rate (%). Degree of liver cell necrosis was assessed on the histopathological specimens. RESULTS The renal parenchymal platinum concentrations (microg/ml) with 4.51+/-2.25 (day 0), 1.59+/-0.70 (day 1), 0.72+/-0.10 (day 3) and 0.20+/-0.06 (day 7) were significantly more pronounced after cisplatin-loaded GMS on days one and three compared to cisplatin with 1.99+/-0.55, 0.08+/-0.03, 0.18+/-0.01 and 0.10+/-0.07, respectively. Relative tumor growth rates resulted in 84.5%+/-26.4 (group I); 241.4%+/-145.1 (II); 331.9%+/-72.2 (III), and 413.6%+/-103.6 (IV) with statistical significant differences between groups I and III, and groups I and IV. Similar degrees of necrosis were observed in both GMSs treated groups, while ballooning of hepatocytes was highest in cisplatin-loaded GMSs. CONCLUSIONS With cisplatin-loaded GMSs more pronounced and prolonged local parenchymal cisplatin concentrations may be achieved offering the advantage of an increased and prolonged anti-cancer effect compared to cisplatin alone or controls. Moreover this proves indirectly the breakdown and release of cisplatin from the GMSs which is of primary importance for drug delivery systems.

Embolization Materials Made of Gelatin: Comparison Between Gelpart and Gelatin Microspheres

Purpose:The object of this study was to assess the level of embolization in the embolized artery and the degradation period of these two embolic agents in the renal arteries using rabbit models.Materials and Methods: The renal artery was embolized using 5 mg of gelatin microspheres (GMSs; diameter, 35–100 μm; group 1) or 1 mg of Gelpart (diameter, 1 mm; group 2). For each group, angiographies were performed on two kidneys immediately after the embolic procedure and on days 3, 7, and 14 after embolization. This was followed by histopathological examinations of the kidneys.Results:Follow-up angiograms on each day revealed the persistence of poorly enhanced wedge-shaped areas in the parenchymal phase in all cases. In group 1, four of six cases showed poorly enhanced small areas in the follow-up angiograms. In group 2, all cases showed poorly enhanced large areas. In the histopathological specimens, it was observed that immediately after embolization, the particles reached the interlobular arteries in group 1 and the interlobar arteries in group 2. In all cases in group 1, the particles were histologically identified even on day 14. In one case in group 2 on day 14, the particles were not identified.Conclusion:In conclusion, although GMSs and Gelpart were similar in the point of gelatin particles, the level of embolization and the degradation period were different between GMSs and Gelpart.

Cisplatin-conjugated degradable gelatin microspheres: fundamental study in vitro

The object of this study was to generate cisplatin-conjugated gelatin microspheres (GMSs) and to confirm the subsequent release of cisplatin in vitro. The GMSs (1 mg) were immersed in 50 microl of a cisplatin solution (0.06, 0.15, 0.27, 0.30 or 0.54 mg ml(-1)) at 38 degrees C to allow conjugation. The cisplatin-conjugated GMSs were then extensively washed in double-distilled water and freeze-dried. The platinum concentration in the GMSs samples was investigated as a function of the concentration of cisplatin solution used in their preparation, the number of immersions in cisplatin (1, 2, 3, 4 or 5) and the period of immersion (1, 6 or 11 h). In vitro release tests were performed at different time intervals (1, 3, 6, 12 or 24 h) to allow the rate of cisplatin release to be calculated. The platinum concentration of the GMSs increased in proportion to the concentration of cisplatin solution and the length or number of immersions in cisplatin. In vitro release tests demonstrate that the release rate (%) from GMSs after 1, 3, 6, 12 or 24 h was 4.8, 5.5, 7.6, 10.0 and 12.4, respectively. We demonstrated the ability of GMSs to bind cisplatin forming cisplatin-conjugated GMSs. Moreover, we showed that cisplatin continued to bind GMSs strongly during the in vitro release test.

Pluronic F127: Application in Arterial Embolization

PURPOSE Pluronic is a substance that is widely used in medical and pharmaceutical fields. In particular, 20% Pluronic F127 solution is a unique substance that is liquid at less than 15 degrees C and gelatinous at 25 to 60 degrees C. In this study, the authors took advantage of the gelation property of Pluronic F127 at human body temperature to simulate embolization and dissolution of the embolism in the renal artery and the superior mesenteric artery (SMA) using a rabbit model. MATERIALS AND METHODS Four female Japanese rabbits (weight, 2.5-3 kg each) were used. The renal artery was fitted with a 4-F cobra-type catheter and embolized with a 20% Pluronic F127 solution at a temperature of 20 degrees C. The embolic effect was evaluated by angiography immediately after the initial injection and every 15 minutes for 2.5 hours after embolization. After 24 hours, pathologic changes of the renal parenchyma were also evaluated. The embolic effect for SMA and ischemic changes of the intestine were evaluated in the same manner. RESULTS Angiographic findings showed that Pluronic F127 caused embolization immediately after injection and dissolved in the renal artery and the SMA after 90 to 120 minutes. The pathologic findings showed no ischemic change in the renal parenchyma. Necrosis was not found in the intestine, but focal hemorrhagic changes were extensively present when the gel had dissolved. This suggested that Pluronic F127 dissolved before severe tissue damage could occur. CONCLUSION Pluronic F127 can potentially be used as a temporary embolic material.

Time-course studies of implanted rabbit VX2 liver tumors to identify the appropriate time for starting hepatic arterial embolization in animal models.

Purpose: We followed the 4-week course of implanted VX2 tumors in rabbits and compared MRI and pathological findings to determine the appropriate time for starting therapy in animal liver tumor models. Materials and Methods: We used 18 Japanese white rabbits. The VX2 liver tumor was harvested from one tumor-bearing rabbit and implanted in the liver of the other 17 rabbits. They were then sacrificed at 1 (n = 5), 2 (n = 3), 3 (n = 4), and 4 weeks (n = 5) after implantation and MRI study. Using MRI scans and/or pathological specimens of individual rabbits, we evaluated the tumor survival ratio, the major tumor axes, intrahepatic metastases, and peritoneal dissemination. Results: All tumor transplantations were successful. At 1 week, 56.25% of the implanted tumors were visualized on MRI scans. At 2 weeks or later, all transplanted rabbits were confirmed to be tumor-bearing on MRI scans. At 3 weeks after implantation, the tumor size was similar on MRI scans and in pathological specimens. There were no intra-hepatic metastases or peritoneal disseminations within 2 weeks of tumor transplantation. Conclusion: We suggest that in studies of implanted VX2 models addressing the treatment of solid hepatic tumors, it may be prudent to start hepatic arterial embolization at 2 weeks after implantation.