Amyloid deposition through endocytosis in vascular endothelial cells.

No mechanistic lead is known for establishing AL amyloid deposits in organs. We here report an electron microscopic (EM) analysis in a case of intestinal AL amyloidosis before initiating treatment for amyloidosis. The dense deposits of amyloid fibrils are concentrated around the small blood vessels in the submucosal area of intestinal tissue. Surprisingly, we observed endothelial cells (ECs) of blood vessels containing plenty of endocytotic (pinocytotic) and transcytotic vesicles at the luminal side and above the basement membrane, indicating the one-way active trafficking of either the immunoglobulin (Ig) light chain or preassembled amyloid fibrils from the luminal side of ECs to the extraluminal area of ECs. Immunoelectron microscopy displayed that the immuno-gold signals were observed in the vascular cavity and the subendothelial area of amyloid deposits. However, there is no sign of an Ig light chain in pinocytotic vesicles. Therefore, the intestinal ECs may actively pump out mainly the preassembled amyloid fibrils (not light chains) from the blood stream into the subendothelial area as a physiologic function.

Hemophagocytic Syndrome-Like Tuberculosis-Immune Reconstitution Inflammatory Syndrome After the Initiation of Hepatic Tuberculosis Treatment.

A 25-year-old Nepalese woman was referred to our hospital because of fever and intermittent abdominal pain with inguinal lymphadenopathy, which had lasted for several months. A liver biopsy of the positron emission tomography-positive lesion led to a diagnosis of hepatic tuberculosis. After the initiation of antituberculosis treatment, her symptoms resolved. However, 11 days after treatment initiation, chest and back pain, high-grade fever, and vomiting appeared and gradually worsened. She developed anemia and her serum ferritin level was elevated. Hemophagocytic syndrome due to the initial deterioration of tuberculosis was suspected and steroid therapy was initiated with the continuation of the antituberculosis drugs. Thereafter, the patient's condition improved remarkably.

[Investigation for bone fixation effect of thin HA coated layer on Ti implants].

In 1995, the author reported the development of unique cementless total hip joints applying hydroxyapatite (HA) flame coating onto a porous titanium surface which is arc-sprayed with pure titanium material in argon gas at atmospheric pressure. The surface roughness of the porous layer was 360 microm Rmax, and the layer was more resistant to blast erosion than the conventional low pressure plasma sprayed porous layers. The thickness of the HA layer was between 20 and 40 microns considering the balance of bone conduction effect of HA and the risk of mechanical detachment of the layers from the porous titanium. Short-term animal tests showed excellent results. At that time a specific question was raised as to whether or not the coated HA layers would remain in the living bone for a long time. The results obtained from observation of retrieved specimens show that the thickness of the HA coating layer is sufficient for new bone formation after implantation, and HA would be absorbed in the body within a few years, after contributing to the new bone formation.

[Development and evaluation of a new surface treatment method for titanium alloy implants].

We developed and evaluated a new method of titanium surface treatment for direct bone fixation of implants. This method consists of hydroxyapatite (HA) flame coating onto a porous titanium surface which is arc-sprayed with pure titanium material in argon gas at atmospheric pressure. The surface roughness of the porous layer was 360 µm, Rmax, and showed higher resistance to blast erosion in comparison with the conventional low-pressure plasma-sprayed porous layers of titanium. The thickness of the HA layer was between 20 and 40 µm considering the balance of bone conduction effect of HA and the possibility of mechanical detachment of the layers from the porous titanium. Short-term animal tests showed excellent results. This new surface treatment method was applied to cementless total hip joints which were used clinically. The results obtained from observations of the retrieved specimens show that the thickness of the HA coating layer is sufficient for the new bone formation after implantation. It was concluded that the new surface treatment method for titanium alloy implants is effective and successful for direct bone fixation.

Assessment of wear in extensively irradiated UHMWPE cups in simulator studies.

Higher levels of UHMWPE crosslinking currently are being advocated for improved wear resistance of acetabular cups. Pioneering Japanese studies, begun in 1971, have achieved good clinical results with UHMWPE irradiated to 1000 kGy for use with a cemented-cup design. The objective of our study was to use contemporary simulator techniques to determine the in vitro wear performance of such high-dose irradiated cups. Extruded UHMWPE cups were processed with 500, 1000, and 1500 kGy of gamma-radiation doses under vacuum, annealed, and machined to shape. The cups were mated with 26-mm alumina heads and run in a multidirectional simulator with bovine serum. Over a 6-million cycle (Mc) study, the weight loss of the nonirradiated control cups averaged 52.8 mg/Mc + 1.4% (wear = 57.2 mm(3)/Mc). In contrast, the irradiated wear cups had a consistent weight gain. Thus cups with irradiation of 500-1500 kGy had no detectable wear in this study. The original machining marks still were partially evident in the wear zones, along with some macrofissures in the 1000- and 1500-kGy cups. Areas adjacent to the fissures showed delaminating plaques of 100-300 microm in size. It also was noted that the wear cups systematically gained more weight than their corresponding soak controls. Each 200-kGy radiation gain increased the fluid sorption ratio by 10%. The increased fluid sorption and evidence of some surface deterioration may indicate that such high-dose irradiated cups are more susceptible to mechanical damage. This indicates that we should take care to ensure that our desire to reduce the wear debris to a zero amount does not result in a modified UHMWPE that lacks the necessary mechanical properties for contemporary metal-backed cup designs.

Thromboresistance of alkali- and heat-treated titanium metal formed with apatite.

Fibrin deposition and platelet adhesion onto alkali- and heat-treated titanium metal (AH-Ti), alkali- and water-treated titanium metal (Wa-Ti), and alkali- and heat-treated titanium metal formed with apatite (Ap-Ti) in simulated body fluid (SBF) were evaluated by exposure to anticoagulated blood or washed platelet suspension (WPS) under static conditions and subsequent observation with scanning electron microscopy (SEM). The results were compared with those for commercially pure titanium metal (cp-Ti). Thrombus formation on AH-Ti and Wa-Ti, which were exposed to heparinized whole blood for 1 h, was significantly less than that on cp-Ti, on which pronounced depositions of fibrin-erythrocytes and lymphocytes were observed. No thrombus was observed on Ap-Ti, possibly because of a high adsorption of heparin. Morphological change of platelets attached to surfaces via adsorbed plasma proteins was found to a significant extent on AH-Ti and Wa-Ti exposed to WPS. However, there was almost no difference between cp-Ti and Ap-Ti in round morphology of adherent platelets. These findings suggested that Ap-Ti exhibits stronger antithrombogenic characteristics than cp-Ti and other materials examined in heparinized blood.

Long-term ex vivo bovine experiments with the Gyro C1E3 centrifugal blood pump.

Centrifugal blood pumps are used widely for cardiopulmonary bypass, as ventricular assist devices, and for extracorporeal membrane oxygenation (ECMO). However, there is no centrifugal blood pump that is suitable for long-term ECMO. The authors developed the Gyro C1E3 centrifugal blood pump (Kyocera Corporation, Kyoto, Japan), which has superior antithrombogenic, antitraumatic, and hydraulic features in comparison with the conventional centrifugal blood pumps. Five ex vivo long-term durability tests of the Gyro C1E3 were performed using healthy miniature calves. The ECMO circuit was composed of a prototype hollow fiber silicone membrane oxygenator and a Gyro C1E3 pump. Venous blood was drained from the left jugular vein of a calf, passed through the oxygenator and infused into the left carotid artery using a Gyro C1E3. Ex vivo studies were performed from 7 to 15 days at a blood flow rate of 1 L/min. During this period, the Gyro C1E3 demonstrated a stable performance without exchanging the pump. Bleeding complications were the major reason for termination of each experiment. Rotational speed was maintained around 2,000 rpm. All five calves demonstrated neither abnormal signs nor abnormal blood examination data throughout the experiment. Neither clot nor thrombus formations were found during the necropsy in the cannula or pump nor were infarctions observed in any of the major organs. In conclusion, the Gyro C1E3 showed a stable and reliable performance during long-term ex vivo bovine experiments under the conditions tested.

Inlet port positioning for a miniaturized centrifugal blood pump.

We are developing the Baylor-Kyocera KP implantable centrifugal blood pump for small sized adult and pediatric patients. This pump eccentrically positions the inlet port, which eliminates flow stagnation around the top pivot bearing. The inlet port design is important because it may vary the inlet orifice pressure on the top housing and change hydraulic performance and hemolytic characteristics. The pressure distribution inside the KP pump was assessed by a computational fluid dynamic (CFD) analysis with 2.7 x 10(5) elements and 3.16 x 10(5) nodes. Hydraulic performance and hemolysis were evaluated with 3 different pump housings, which had 3.8, 4.5, and 6.1 mm offset inlet ports from the center in a mock circuit. The CFD analysis revealed that the pressure gradually increased from the center toward the peripheral. The pressure difference between the 3.8 to 6.1 mm offsets was less than 600 Pa. The hydraulic performance did not drastically change at 3.8, 4.5, and 6.1 mm offset from the center. However, the hemolysis increased with the increase of the port offset from 0.0080+/- 0.0048 to 0.054 +/- 0.028 g/100 L. The inlet port positioning is important to attain less blood trauma in this small Gyro centrifugal blood pump. The preferable position of the inlet port is less than 4.5 mm offset from the center.

Impeller inner diameter in a miniaturized centrifugal blood pump.

To design a miniaturized centrifugal blood pump, the impeller internal diameter (ID), which is a circle diameter on the inner edge of the vane, is considered one of the important aspects. Hydraulic performance, hemolysis, and thrombogenicity were evaluated with different impeller IDs. Two impellers were fabricated with an outer diameter of 35 mm, of which 1 had an 8 mm ID impeller and the other had a 12 mm ID. These impellers were combined with 2 different housings in which the inlet port was eccentrically positioned 3.8 and 4.5 mm offset from the center. The hydraulic performance and hemolysis were evaluated in a mock circuit, and thrombogenicity was evaluated in a 2 day ex vivo study with each impeller housing combination. Both impellers required 3,000 rpm in the 3.8 mm offset inlet to attain 5 L/min against 100 mm Hg (left ventricular assist device condition). The 8 mm ID impeller required 3,200 rpm, and the 12 mm ID impeller required 3,100 rpm in the 4.5 mm offset housing. The normalized index of hemolysis was 0.0080 +/- 0.0048 g/100 L in the 8 mm ID impeller with the 3.8 mm offset and 0.022 +/- 0.018 g/100 L with 4.5 mm offset. The 12 mm ID impeller had 0.068 +/- 0.028 g/100 L with the 3.8 mm offset and 0.010 +/- 0.002 g/100 L with the 4.5 mm offset. After the 2 day ex vivo study, no blood clot was formed around the top bearing in all the pump heads. The 8 mm ID impeller with 3.8 mm offset inlet and the 12 mm ID impeller with the 4.5 mm offset had less hemolysis compared to the other pump heads that were subjected to 14 day ex vivo and 10 day in vivo studies. The 8 mm ID impeller with the 3.8 mm offset inlet had a blood clot around the top bearing after the 14 day ex vivo study. No thrombus was found around the top bearing of the 12 mm ID impeller with the 4.5 mm offset in the 10 day in vivo study. These results suggest that the ID does not greatly change the hydraulic performance of a small centrifugal blood pump. The proper combination of the impeller ID and inlet port offset obtains less hemolysis. The larger impeller ID is considered to have less thrombogenicity around the top bearing.