Three-Dimensional Bio-Printed Tubular Tissue Using Dermal Fibroblast Cells as a New Tissue-Engineered Vascular Graft for Venous Replacement.

The current study was a preliminary evaluation of the feasibility and biologic features of three-dimensionally bio-printed tissue-engineered (3D bio-printed) vascular grafts comprising dermal fibroblast spheroids for venous replacement in rats and swine. The scaffold-free tubular tissue was made by the 3D bio-printer with normal human dermal fibroblasts. The tubular tissues were implanted into the infrarenal inferior vena cava of 4 male F344-rnu/rnu athymic nude rats and the short-term patency and histologic features were analyzed. A larger 3D bio-printed swine dermal fibroblast-derived prototype of tubular tissue was implanted into the right jugular vein of a swine and patency was evaluated at 4 weeks. The short-term patency rate was 100%. Immunohistochemistry analysis showed von Willebrand factor positivity on day 2, with more limited positivity observed on the luminal surface on day 5. Although the cross-sectional area of the wall differed significantly between preimplantation and days 2 and 5, suggesting swelling of the tubular tissue wall (both p < 0.01), the luminal diameter of the tubular tissues was not significantly altered during this period. The 3D bio-printed scaffold-free tubular tissues using human dermal or swine fibroblast spheroids may produce better tissue-engineered vascular grafts for venous replacement in rats or swine.

Novel Screening System for Biliary Excretion of Drugs Using Human Cholangiocyte Organoid Monolayers with Directional Drug Transport.

It has recently been reported that cholangiocyte organoids can be established from primary human hepatocytes. The purpose of this study was to culture the organoids in monolayers on inserts to investigate the biliary excretory capacity of drugs. Cholangiocyte organoids prepared from hepatocytes had significantly higher mRNA expression of CK19, a bile duct epithelial marker, compared to hepatocytes. The organoids also expressed mRNA for efflux transporters involved in biliary excretion of drugs, P-glycoprotein (P-gp), multidrug resistance-associated protein 2 (MRP2), and breast cancer resistance protein (BCRP). The subcellular localization of each protein was observed. These results suggest that the membrane-cultured cholangiocyte organoids are oriented with the upper side being the apical membrane side (A side, bile duct lumen side) and the lower side being the basolateral membrane side (B side, hepatocyte side), and that each efflux transporter is localized to the apical membrane side. Transport studies showed that the permeation rate from the B side to the A side was faster than from the A side to the B side for the substrates of each efflux transporter, but this directionality disappeared in the presence of inhibitor of each transporter. In conclusion, the cholangiocyte organoid monolayer system has the potential to quantitatively evaluate the biliary excretion of drugs. The results of the present study represent an unprecedented system using human cholangiocyte organoids, which may be useful as a screening model to directly quantify the contribution of biliary excretion to the clearance of drugs.

The comparison of the clinical outcomes and costs between surgical aortic valve replacement and transcatheter aortic valve implantation based on the Japanese Diagnosis Procedure Combination database.

PURPOSE: We evaluated the clinical outcomes and costs of surgical aortic valve replacement (SAVR) and transfemoral transcatheter aortic valve implantation (TAVI) for aortic stenosis using the Japanese Diagnosis Procedure Combination (DPC) database. METHODS: Using our extraction protocol, we retrospectively analyzed summary tables in the DPC database from 2016 to 2019, which were provided by the Ministry of Health, Labor and Welfare. A total of 27,278 patients were available (SAVR, n = 12,534; TAVI, n = 14,744). RESULTS: The TAVI group was older than the SAVR group (SAVR vs. TAVI: 74.6 vs. 84.5 years; P < 0.01), with a lower in-hospital mortality (1.0 vs. 0.6%; P < 0.01) and a shorter hospital stay (26.9 vs. 20.3 days; P < 0.01). TAVI conferred more total medical service reimbursement points than SAVR (493,944 vs. 605,241 points; P < 0.01), especially materials points (147,830 vs. 434,609 points; P < 0.01). Total insurance claims for TAVI were approximately 1 million yen higher than those for SAVR. Regarding the details of costs, only the operation cost was higher, while other costs were lower with TAVI than with SAVR. CONCLUSION: Our analysis revealed that both SAVR and TAVI showed acceptable clinical outcomes. TAVI was associated with higher total insurance claims than SAVR. If the material cost of TAVI operations can be reduced, greater cost-effectiveness can be expected.

[Surgical Left Atrial Ablation Through the Left Atrial Appendage During Aortic Valve Replacement].

This is a 2-case report of concomitant left atrial( LA) surgical ablation through the left atrial appendage( LAA) for atrial fibrillation( AF) during aortic valve replacement( AVR). Case 1, a 74-year-old man, and Case 2, an 85-years-old woman, were presented for AVR and AF. Under the cardio-pulmonary bypass, right and left pulmonary vein isolations( PVIs) were performed with radiofrequency( RF) ablation devices. Through the opened LAA, an RF lesion was placed to connect the bilateral PVIs and the LAA, and another RF line was placed toward the mitral annulus. A right atrial( RA) isthmus ablation was also performed through a RA incision. Postoperatively, sinus rhythm and both RA and LA contraction were obtained, and have been maintained for 7.5 and 5 years without classⅠ/Ⅲ antiarrhythmic drugs in Case 1 and 2, respectively. This procedure is considered to be effective and safe, although further study is needed.

Scaffold-free human vascular calcification model using a bio-three-dimensional printer.

Morbidity and mortality rates associated with atherosclerosis-related diseases are increasing. Therefore, developing new research models is important in furthering our understanding of atherosclerosis and investigate novel treatments. Here, we designed novel vascular-like tubular tissues from multicellular spheroids composed of human aortic smooth muscle cells, endothelial cells, and fibroblasts using a bio-3D printer. We also evaluated their potential as a research model for Mönckeberg's medial calcific sclerosis. The tubular tissues were sufficiently strong to be handled 1 week after printing and could still be cultured for 3 weeks. Histological assessment showed that calcified areas appeared in the tubular tissues within 1 week after culture in a medium containing inorganic phosphate (Pi) or calcium chloride as the calcification-stimulating factors. Calcium deposition was confirmed using micro-computed tomography imaging. Real-time quantitative reverse transcription polymerase chain reaction analysis revealed that the expression of osteogenic transcription factors increased in calcified tubular tissues. Furthermore, the administration of Pi and rosuvastatin enhanced tissue calcification. The bio-3D printed vascular-like tubular structures, which are composed of human-derived cells, can serve as a novel research model for Mönckeberg's medial calcific sclerosis.

How to select the optimal size of frozen elephant trunk in total arch replacement for type A acute aortic dissection.

BACKGROUND: Total arch replacement (TAR) with a frozen elephant trunk (FET) is a common technique for acute aortic dissection, but there is no consensus on the optimal size of the FET. METHODS: Forty-four patients who underwent TAR with FET for acute aortic dissection at our hospital since 2014 were included. The aortic diameter obtained from FET was measured on postoperative computed tomography (CT) and the estimated oversizing ratio was calculated. We investigated the relationship between the estimated oversizing ratio and postoperative outcomes. We also measured the maximum true lumen diameter, circumference of the true lumen, and total aortic diameter at the same level as the FET end on preoperative CT and examined the correlation with the aortic diameter obtained from FET. RESULTS: The average estimated oversizing ratio was 109%. Early postoperative CT showed complete thrombosis of the false lumen in 41 (93.2%) patients. No distal stent graft-induced new entry occurred during follow-up. The correlation coefficients between the three measurements and aortic diameter obtained from FET were 0.64 (maximum true lumen diameter), 0.76 (true lumen diameter calculated from circumference), and 0.72 (total aortic diameter), respectively. CONCLUSIONS: The aortic diameter obtained from FET on postoperative CT was strongly correlated with the true lumen diameter calculated from the circumference and total aortic diameter on preoperative CT. It is reasonable to select a size of approximately 130% of the true lumen diameter calculated from the circumference or 80% to 85% of the total aortic diameter.

[Tricuspid Valve Ring Annuloplasty with Posterior Annular Plication for Functional Tricuspid Regurgitation].

This report presents a modified procedure of tricuspid valve ring annuloplasty (R-TAP) with posterior annular plication for functional tricuspid regurgitation (TR). Sutures on the native annulus were placed by a standard fashion in R-TAP, and those on the posterior annulus and its bilateral commissures were passed through in a narrow range between the 3 and 4 o'clock positions of the 26-mm ring. The other sutures were done with an usual manner and the ring was fixed to the annulus, resulting in the posterior annular plication( bicuspidization). Follow-up was performed for more than 5 years( mean: 7.9 years, range:5.5~11.5 years) by echocardiography in 13 cases. Postoperative TR reduced significantly to less than moderate, which was maintained during the entire follow-up period, even in the case with atrial fibrillation. There was no sign of tricuspid stenosis. R-TAP with posterior annular plication was feasible, reproducible, and effective, although further investigation is needed.

Directional Drug Transport through Membrane-Supported Monolayers of Human Liver-Derived Cell Lines.

The aim of this work is to develop a new assay system for screening biliary excretion drugs. When monolayers of human liver-derived cell lines HepG2 and Huh-7 were grown on an insert membrane, the efflux ratio (ER: ratio of the apparent permeability coefficient in the basal-to-apical direction (Papp,B-to-A) to that in the apical to basal direction (Papp,A-to-B)) of sulfobromophthalein (BSP), a model substrate of multidrug resistance-associated protein 2 (MRP2), was greater than 1.0, indicating transport of BSP in the efflux direction. The efflux transport was significantly suppressed by MK-571, an inhibitor of MRPs, in both cell lines. Expression of MRP2 mRNA in HepG2 and Huh-7 was 3.5- and 1.4-fold higher, respectively, than in primary human hepatocytes, while expression of P-glycoprotein and breast cancer resistance protein mRNAs was markedly lower, supporting the idea that MRP2 is the main mediator of directional BSP transport in this assay system. The advantage of our system is the potential to quantitatively evaluate biliary excretion of MRP2 substrates in vitro.

[Repeated Heart Failure after Transcatheter Aortic Valve Implantation Using Self-expandable Bioprosthetic Valve:Report of a Case].

The patient was an 81-year-old man. Transcatheter aortic valve implantation( TAVI) was performed for severe aortic stenosis using Evolut R. The patient moved to intensive care unit without an adverse event after the operation. But repeated acute heart failure occurred several times during hospital stay. Mitral regurgitation (MR) was worsened from mild at baseline to moderate or more by transthoracic echocardiography. Various factors that worsened MR after TAVI have been reported, and treatment strategy for severe aortic stenosis patients with MR should be carefully developed.

Early results of expanding the anatomical indications for using a Gore Iliac branch endoprosthesis to treat aortoiliac and iliac aneurysms.

PURPOSE: To assess the safety and anatomical suitability of using a Gore Iliac Branch Endoprosthesis (IBE) in aortoiliac and iliac aneurysm repair. METHODS: Between 2017 and 2020, 20 patients underwent endovascular aneurysm repair (EVAR) with a Gore IBE device (bilateral IBE, n = 1) after expanding the instructions for use (IFU) criteria. We evaluated the early clinical outcomes and suitability of the IFU criteria, retrospectively. RESULTS: Six patients (30%) met all the IFU criteria. Anatomical suitability according to the IFU criteria for the collective total of 21 IBE limbs was confirmed for 10 (47.6%) proximal common iliac arteries, 21 (100%) external iliac arteries, 18 (85.7%) internal iliac arteries, and in the length from the lowest renal artery to the iliac bifurcation in 15 (71.8%) patients. Assisted primary technical success was achieved in all patients with various bail-out techniques. One patient (5%) required a bare-stent insertion 7 days after EVAR for severe stenosis in the ipsilateral limb caused by a small terminal aorta. There was no case of occlusion of an iliac branch component device. CONCLUSIONS: Gore IBEs were implanted safely and effectively with various bail-out techniques to repair aortoiliac and iliac aneurysms in our Japanese patients with a low rate of inclusion IFU criteria.

Correction: Fabrication of scaffold-free tubular cardiac constructs using a Bio-3D printer.

[This corrects the article DOI: 10.1371/journal.pone.0209162.].

[Preservation of the Spinal Perfusion Artery during Descending Thoracic and Thoracoabdominal Aortic Surgery].

This is a 3-case report of successful descending thoracic and thoracoabdominal aortic surgery by preserving the spinal perfusion artery (SPA) identified preoperatively. In Case 1, an 85-year-old woman, computed tomography (CT) showed Crawford type Ⅴ thoracoabdominal aortic aneurysm (TAAA:60 mm) and a SPA originated from L2. In Case 2, a 76-year-old man, CT revealed type Ⅳ TAAA( 58 mm) and a SPA originated from Th11. In Case 3, a 74-year-old man, CT detected an infectious pseudoaneurysm(44 mm) in the descending thoracic aorta with 2 SPAs originating from Th10 and L2. The ranges of graft replacement were Th7-Th12, Th12-L4, and Th8-Th10, respectively, while preserving all SPAs. All patients recovered well without postoperative neurological deficits. Although the protective effect of the SPA preservation against the spinal cord ischemia is still controversial, preoperative identification of the SPA was useful for planning a surgical strategy for descending thoracic and thoracoabdominal aortic repair surgery.

Patient-derived ovarian cancer organoids capture the genomic profiles of primary tumours applicable for drug sensitivity and resistance testing.

The use of primary patient-derived organoids for drug sensitivity and resistance testing could play an important role in precision cancer medicine. We developed expandable ovarian cancer organoids in < 3 weeks; these organoids captured the characteristics of histological cancer subtypes and replicated the mutational landscape of the primary tumours. Seven pairs of organoids (3 high-grade serous, 1 clear cell, 3 endometrioid) and original tumours shared 59.5% (36.1-73.1%) of the variants identified. Copy number variations were also similar among organoids and primary tumours. The organoid that harboured the BRCA1 pathogenic variant (p.L63*) showed a higher sensitivity to PARP inhibitor, olaparib, as well as to platinum drugs compared to the other organoids, whereas an organoid derived from clear cell ovarian cancer was resistant to conventional drugs for ovarian cancer, namely platinum drugs, paclitaxel, and olaparib. The overall success rate of primary organoid culture, including those of various histological subtypes, was 80% (28/35). Our data show that patient-derived organoids are suitable physiological ex vivo cancer models that can be used to screen effective personalised ovarian cancer drugs.

Drug response analysis for scaffold-free cardiac constructs fabricated using bio-3D printer.

Cardiac constructs fabricated using human induced pluripotent stem cells-derived cardiomyocytes (iPSCs-CMs) are useful for evaluating the cardiotoxicity of and cardiac response to new drugs. Previously, we fabricated scaffold-free three-dimensional (3D) tubular cardiac constructs using a bio-3D printer, which can load cardiac spheroids onto a needle array. In this study, we developed a method to measure the contractile force and to evaluate the drug response in cardiac constructs. Specifically, we measured the movement of the needle tip upon contraction of the cardiac constructs on the needle array. The contractile force and beating rate of the cardiac constructs were evaluated by analysing changes in the movement of the needle tip. To evaluate the drug response, contractile properties were measured following treatment with isoproterenol, propranolol, or blebbistatin, in which the movement of the needle tip was increased following isoproterenol treatment, but was decreased following propranolol or blebbistain, treatments. To evaluate cardiotoxicity, contraction and cell viability of the cardiac constructs were measured following doxorubicin treatment. Cell viability was found to decrease with decreasing movement of the needle tip following doxorubicin treatment. Collectively, our results show that this method can aid in evaluating the contractile force of cardiac constructs.

Placement of a Zenith® Dissection Endovascular System in the Descending Thoracic Aorta Can Hamper Further Surgical Aortic Operations.

INTRODUCTION: The efficacy of endovascular treatment for complicated Stanford type B acute aortic dissection is being established. However, aortic events sometimes occur, and some cases require surgical intervention. REPORT: A 52 year old man underwent ascending aorta replacement for Stanford type A acute aortic dissection in August 2016. Post-operative computed tomography (CT) showed residual dissection from the aortic arch to the right common iliac artery and a large re-entry in the right common iliac artery (RCIA). Two months after the operation, CT revealed enlargement of the false lumen of the thoracic aorta and the thoracic aortic diameter. Aiming to reduce the false lumen and remodel the aorta, a three stage operation was performed, as described below. Four months after the dissection, total aortic arch replacement and a frozen elephant trunk insertion were performed as the first stage. Subsequently, as a second stage operation, thoracic endovascular repair (TEVAR) was performed using a Zenith® Dissection Endovascular System (Cook Japan Co., Ltd, Tokyo, Japan), with the aim of expanding the true aortic lumen. The implanted devices were a stent graft for the proximal part and two bare stents for the middle and distal part. As a third stage operation, abdominal aortic endovascular treatment was performed with the purpose of closing the re-entry from the RCIA. However, two years after the three stage operation, CT showed that the thoracic aorta was over 60 mm in diameter. Graft replacement of the thoraco-abdominal aorta was performed. The bare stents were expected to be easily removable from the aorta, but unexpectedly, they were strongly attached to the intima, which made it extremely difficult to perform surgical and aortic operations. DISCUSSION: Surgical operations for the aorta can become more difficult after bare stent placement in the aorta.

Establishment and characterization of a new malignant peritoneal mesothelioma cell line, KOG-1, from the ascitic fluid of a patient with pemetrexed chemotherapy resistance.

Malignant peritoneal mesothelioma (MPeM) is a rare and aggressive form of malignant mesothelioma. Sufficient biological tools for studying the functional characteristics of this cancer have not been developed. Therefore, in this study, a novel human cancer cell line, KOG-1, was established from ascites fluids isolated from a 39-year-old Japanese woman with pemetrexed-resistant MPeM. Cells were dendritic or linear immediately after thawing, showed a jigsaw puzzle-like and spindle arrangement during growth, and formed monolayers without contact inhibition in two-dimensional (2D) culture. The population doubling time was 13.7 h. Karyotypic and molecular genetic analyses showed that chromosome numbers ranged from 62 to 142, with a peak of 73 with complicated copy number alterations. No germline BAP1 pathogenic variant was detected. Cells expressed various tumor markers of mesothelioma, such as calretinin, podoplanin, and Wilms tumor 1 (WT-1). Drug sensitivity and resistance testing with a set of 36 drugs using 2D and three-dimensional (3D) culture models demonstrated that KOG-1 cells showed high and low sensitivity to pemetrexed under 2D and 3D culture conditions, respectively, whereas control ovarian cancer cell lines showed low sensitivity to pemetrexed under both culture conditions. This newly established cell line will be a valuable biological resource to expand the feasibility of functional studies as well as drug testing for potential therapeutic purposes in MPeM.

[Retrosternal Giant Aortic Aneurysm;Report of Two Cases].

This is a 2-case report of successful aortic repair surgery for the retrosternal giant aortic aneurysm. Our surgical strategy is "deep hypothermia and left ventricular( LV) unloading under cardiopulmonary bypass before approaching to the aortic aneurysm" in case of possible catastrophic bleeding. Case 1, a 64-year-old woman, had a retrosternal pseudoaneurysm (80 mm) at the distal anastomosis of a Dacron graft used to replace the ascending aorta 7 years before. An LV vent tube was cannulated via the right upper pulmonary vein through an inferior T-shaped ministernotomy. Case 2, an 86-year-old woman, had a retrosternal chronic aortic dissecting aneurysm (66 mm). An LV vent cannula was inserted via the LV apex through a left minithoracotomy. Arch replacement and ascending aorta replacement were performed in Case 1 and 2, respectively, without cardiac, neurological, or any other complications. This strategy is safe and useful in a case with complex aortic disease.

[Mitral Valve Replacement with the On-X Valve].

This is a 3-case report of mitral valve replacement (MVR) with an On-X mechanical valve followed up to 10 years. Case 1(64-year-old man) and case 2 (66-year-old woman) experienced traffic accident and traumatic event, respectively, in their chronic phase after MVR. Case 1 had multiple bone fractures of the bilateral lower limbs, which was followed by systemic infection and pyogenic spondylitis. He needed long-term antibiotics therapy for more than 4 years. Case 2 fell down at home and severely hit her head, which resulted in a traumatic subarachnoid hemorrhage. She was in a deep coma, and needed discontinuation of anticoagulation therapy for 4 weeks. Case1, 2, and 3(54-year-old man) are doing well in New York Heart Association functional class I without any valve-related thromboembolic or hemorrhagic events at 10, 9 and 8 years after MVR, respectively. On-X valve performance has also been found well maintained in all cases by echocardiography, even after traumatic accident or discontinuation of anticoagulation in Case 1 and 2. In this report, the On-X mechanical valve demonstrated good midterm result of its valve performance in the mitral position and its potential advantages in antithrombogenicity.

Adult-onset primary Ewing's sarcoma of the right atrium: a case report.

BACKGROUND: Primary cardiac tumors, which are only detected in 0.001-0.03% of autopsies, are rare. Only 25% of primary cardiac tumors are malignant, of which 95% are sarcomas. Ewing's sarcoma, one of the Ewing's sarcoma of family tumors, is thought to be derived from neural crest cells. While Ewing's sarcoma usually presents in the bone of children, Ewing's sarcoma of cardiac origin is rare, with only a few reports described in the literature. The prognosis is unpredictable because of the scarcity and unestablished treatment. We herein report an extremely rare case of primary cardiac Ewing's sarcoma in the right atrium of a 64-year-old man. CASE PRESENTATION: The patient is a 64-year-old Japanese male who was referred to our hospital to treat a floating mass of the right atrium (RA). Although the patient was asymptomatic, we performed an operation to urgently resect the floating mass on the next day of admission due to the risk of pulmonary embolism. The operation was performed under cardiopulmonary bypass and cardiac arrest. We resected the tumor with at least 1.5 cm of the RA wall as a margin. The postoperative pathological diagnosis of the mass was compatible with a primitive neuroectodermal tumor (PNET, a form of Ewing's sarcoma). The cells were positive for CD56, CD99, and Vimentin and negative for S-100 and Desmin. Although no malignant cells were observed in the margin of the resected RA wall and the sarcoma was completely resected, he was transferred to another hospital to receive adjuvant postoperative chemotherapy to improve the prognosis by preventing subclinical micrometastasis. CONCLUSIONS: We experienced an extremely rare case of primary cardiac Ewing's sarcoma in the right atrium of a 64-year-old man, which was successfully resected under cardiac arrest. Although the sarcoma was completely resected, postoperative chemotherapy and long-term follow-up are recommended for patients with primary cardiac sarcoma because of the high rates of metastasis and recurrence.

Publisher Correction: Development of an immunodeficient pig model allowing long-term accommodation of artificial human vascular tubes.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.