Chondroinduction of mouse mesenchymal stem cells in three-dimensional highly porous matrix scaffolds

Porous polyvinyl formal (PVF) resin and poly(lactide-caprolactone) [P(LA/CL)] sponges were examined as three-dimensional matrices for chondroinduction of cultured bone marrow mesenchymal stem cells (MSCs). Approximately 5 x 10(5) mouse MSCs were seeded in porous PVF resin or P(LA/CL) sponges and were cultured for up to 1 month in serum-free high-glucose Dulbecco's modified Eagle's medium containing 10 ng/mL transforming growth factor-beta3 and 100 nM dexamethasone for chondroinduction. After the 1-month culture period, the PVF resin and P(LA/CL) sponges contained approximately twice the amount of glycosaminoglycans compared with the control pellet. Safranin-O staining of PVF and P(LA/CL) after 1 month of culture revealed a cartilage-like extracellular matrix containing glycosaminoglycans and collagen. When implanted into nude mice, PVF and P(LA/CL) seeded with MSCs were found to be both biocompatible and chondroinductive. These highly porous scaffolds can maintain a large number of cells in a three-dimensional structure. Both are potentially promising for the chondroinduction of bone marrow MSCs for research and clinical applications.     

Efficient proliferation and maturation of fetal liver cells in three-dimensional culture by stimulation of oncostatin M, epidermal growth factor, and dimethyl sulfoxide

For the purpose of applying fetal liver cells (FLCs) as a cell source to tissue-engineered bioartificial livers, three-dimensional (3-D) cultures of FLCs using a porous polymer scaffold, as well as monolayer cultures as a control, were simultaneously performed. To achieve efficient growth and differentiation, the FLCs were cultured in the growth medium for the first 3 weeks and then cultured in the differentiation medium for 3 more weeks. In these cultures, stimulating factors (oncostatin M (OSM), epidermal growth factor (EGF), hepatocyte growth factor (HGF), or dimethyl sulfoxide (DMSO)) were added to the media, and their effects were examined. When the growth medium containing OSM and EGF was used, EGF stimulated the growth of FLCs synergistically with OSM. For the differentiation of FLCs into mature hepatocytes, DMSO added to the differentiation medium remarkably enhanced albumin secretion in the 3-D and monolayer cultures, although HGF was effective only in the monolayer culture. Microscopic observation proved that FLCs exhibited hepatocyte-like morphology only in the media containing DMSO. In conclusion, successive supply of the growth medium containing EGF and OSM and the differentiation medium containing DMSO efficiently induced the growth of the 3-D cultured FLCs and their differentiation into mature hepatocytes.     

Pathological and immunohistological findings and genetic aberrations of intestinal enteropathy-associated T cell lymphoma in Japan

Takeshita M, Nakamura S, Kikuma K, Nakayama Y, Nimura S, Yao T, Urabe S, Ogawara S, Yonemasu H, Matsushita Y, Karube K & Iwashita A
(2011) Histopathology 58 , 395–407
Pathological and immunohistological findings and genetic aberrations of intestinal enteropathy‐associated T cell lymphoma in Japan Aims: To elucidate the clinicopathological findings of primary intestinal enteropathy‐associated T cell lymphoma (EATL) in Japan, a non‐endemic area for coeliac disease. Methods and results: Of the 24 cases, four (17%) had large‐cell lymphoma (type I), and the remaining 20 (83%) had medium‐sized lymphoma (type II). Lymphoma cells of the three type I cases were CD56‐positive. Only one (4%) case showed typical CD56‐ and CD8‐negative and CD30‐positive type I EATL. In type II EATL, lymphoma cells of the 16 (80%) and 11 (55%) cases were positive for CD56 and CD8, respectively. Intramucosal tumour spreading and adjacent enteropathy‐like lesions were detected in 15 (71%) and 16 (76%) of 21 cases, with a severe increase of intraepithelial lymphocytes (IELs) in 12 (57%). IELs of enteropathy‐like lesions in five (24%) cases expressed T‐bet, with no cases of CD30‐positive IELs. Characteristic findings from comparative genomic hybridization of 15 cases indicated gains of 8q2 (47%), Xp (53%) and Xq (73%), but no gain of 9q3. Regarding, human leucocyte antigen (HLA) status, six cases examined did not express the DQB1*02 allele. Conclusions: Japanese EATL exhibited different histology, cytogenetic findings and HLA status from those of typical type I EATL. The rare incidence of coeliac disease may influence the tumour cell characteristics of EATL and IELs.     

Polyvinyl alcohol scaffolds and supplementation support 3D and sphere culturing of human cancer cell lines by reducing apoptosis and promoting cellular proliferation

Three-dimensional (3D) culturing mimics the heterogeneous cellular conditions of the in vivo tumor microenvironment compared to 2D monolayer-cultured cells and 3D cultures of established cancer cell lines (sphere culture) or patient-derived cancer cells (organoid culture) are frequently used for cancer research or drug screening and evaluation. To establish more cost and time-efficient 3D culture methods for cancer cell lines, we supplemented sphere culture medium with polyvinyl alcohol (PVA) and found that 3D sphere cultures of breast and pancreatic cancer cell lines were significantly increased. Mechanistically, we found that PVA prevented cell death and promoted cellular proliferation while maintaining levels of stemness-related gene expression. Furthermore, we showed that polyvinyl formal resin (PVF) 3D scaffolds made by cross-linked PVA can function in serum-free, long-term 3D cultures to support maintenance of sphere- or tumor-like cell masses for diverse cancer cell types. Taken together, we demonstrate the effectiveness of PVA and PVF in human cancer cell line culture protocols.     

Central circulatory and peripheral O<Subscript>2</Subscript> extraction changes as interactive facilitators of pulmonary O<Subscript>2</Subscript> uptake during a repeated high-intensity exercise protocol in humans

It has frequently been demonstrated that prior high-intensity exercise facilitates pulmonary oxygen uptake response at the onset of subsequent identical exercise. To clarify the roles of central O₂ delivery and/or peripheral O₂ extraction in determining this phenomenon, we investigated the relative contributions of cardiac output (CO) and arteriovenous O₂ content difference to the transient during repeated bouts of high-intensity knee extension (KE) exercise. Nine healthy subjects volunteered to participate in this study. The protocol consisted of two consecutive 6-min KE exercise bouts in a supine position (work rate 70–75% of peak power) separated by 6 min of rest. Throughout the protocol, continuous-wave Doppler ultrasound was used to measure beat-by-beat CO (i.e., via simultaneous measurement of stroke volume and the diameter of the arterial aorta). The phase II response was significantly faster and the slow component (phase III) was significantly attenuated during the second KE bout compared to the first. This was a result of increased CO during the first 30 s of exercise: CO contributing to 100 and 56% of the speeding at 10 and 30 s, respectively. After this, the contribution of became increasingly more predominant: being responsible to an estimated 64% of the speeding at 90 s, which rose to 100% by 180 s. This suggests that, while both CO and clearly interact to determine the response, the speeding of kinetics by prior high-intensity KE exercise is predominantly attributable to increases in .     

Combination tumor immunotherapy with radiotherapy and Th1 cell therapy against murine lung carcinoma

Mice bearing established Lewis lung carcinoma (LLC) expressing model tumor antigen, ovalbumin (OVA) (LLC-OVA) marginally responded to local radiotherapy, but none of the mice was cured. In contrast, treatment of the tumor-bearing mice with intratumoral injection of tumor-specific T helper type 1 (Th1) cells and tumor antigen (OVA) after radiotherapy dramatically prolonged the survival days and induced complete cure of the mice at high frequency (80%). Radiation therapy combined with Th1 cells or OVA alone showed no significant therapeutic activity against LLC-OVA. Such a strong therapeutic activity was not induced by intratumoral injection of Th1 cells plus OVA. Compared with other treatment, radiation therapy combined with Th1 cells and OVA was superior to induce the generation of OVA/H-2^b tetramer⁺ tumor-specific cytotoxic T lymphocyte (CTL) with a strong cytotoxicity against LLC-OVA in draining lymph node (DLN). Moreover, the combined therapy is demonstrated to inhibit the growth of tumor mass, which grew at contralateral side. These results indicated that radiotherapy combined with Th1 cell/vaccine therapy induced a systemic antitumor immunity. These findings suggested that combination therapy with radiotherapy and Th1 cell/vaccine therapy may become a practical strategy for cancer treatment. Hiroshi Yokouchi and Kenji Chamoto are equally contributed.     

Stroke patients' evolving symptoms assessed by tractography

PURPOSE: To test the hypothesis that an interval evolution in the location of the depicted sensorimotor tract relative to the infarct (the "tract-infarct relationship") may be related to stroke victims' symptom progression. MATERIALS AND METHODS: Patients (N = 7) who underwent multiple diffusion-tensor imaging (DTI) studies during symptomatic progression were included in this study. DTI was performed using a single-shot echo-planar imaging (EPI) technique with a motion-probing gradient in six orientations, a b-value of 800 seconds/mm2, and six image averages. The total scan time was four minutes and 24 seconds. Fiber-tracking of the sensorimotor pathways was performed, and the locations of these tracts were retrospectively assessed in relation to the evolution of the symptoms. RESULTS: Five of the seven patients showed an interval enlargement of the infarct on diffusion-weighted (DW) images. In two of these cases the lesion enlarged to involve the sensorimotor tracts, while in three cases the lesion enlarged only so far to come into close proximity to the sensorimotor tract. In the remaining two cases there was no interval enlargement of the infarct, and therefore the tract-infarct relationship could not account for the evolution of the symptoms in these cases. CONCLUSION: A tract-infarct relationship can be observed with the use of a fiber-tracking technique, and the results may improve our understanding of the symptom progression seen in stroke victims.     

Oculomotor nerve palsy evaluated by diffusion-tensor tractography

Introduction The aim of the study was to test the feasibility of the tractography technique based on diffusion-tensor imaging (DTI) for the assessment of small infarcts involving the brainstem.Methods A patient who presented with an isolated left third cranial nerve palsy underwent magnetic resonance examination. Images were obtained by use of a whole-body, 1.5-T imager. Data were transferred to an off-line workstation for fiber tracking.Results The conventional diffusion-weighted imaging (DWI) performed using a 5 mm slice thickness could only depict an equivocal hyperintensity lesion located at the left paramedian midbrain. An additional thin-slice DTI was performed immediately after the initial DWI using a 3 mm slice thickness and was able to delineate the lesion more clearly. Image postprocessing of thin-slice DTI data revealed that the lesion location involved the course of the third cranial nerve tract, corresponding with the patient’s clinical symptoms.Conclusion The tractography technique can be applied to assess fine neuronal structures of the brainstem, enabling direct clinicoradiological correlation of small infarcts involving this region.     

Optimization of TI values in inversion-recovery MR sequences for the depiction of fine structures within gray and white matter: separation of globus pallidus interna and externa

RATIONALE AND OBJECTIVES: The authors' purpose was to search the inversion time (TI) values that enable the best differentiation of fine structures in gray matter (gray-gray differentiation). MATERIALS AND METHODS: Seven healthy adult volunteers with no history of neurologic disease or head trauma were recruited and gave their informed consent. The subjects consisted of two men and five women ranging in age from 25 to 38 years, with a mean age of 28 years +/- 5. The subjects were imaged with a turbo spin-echo inversion-recovery sequence. This sequence was performed in the axial plane at the level of the basal ganglia with the following parameters: repetition time, 3,200 msec; echo time, 15 msec; three signals acquired; echo train, seven; section thickness, 3 mm; matrix size, 256 x 256; and field of view, 180 mm. The tested values were TI = 100, 200, 300, 400, and 500 msec. Region-of interest measurements were performed at the following anatomic structures and represent gray-gray and white-white differentiations, respectively: globus pallidus externa versus globus pallidus interna, and optic radiation versus surrounding white matter. RESULTS: The maximum contrast index value occurred at TI = 400 msec for globus pallidus externa versus globus pallidus interna (P < .05) With the contrast-to-noise ratio, no significant difference in gray-gray differentiation was observed among the various TIs. The minimum signal-to-noise ratio of the gray matter occurred at TI = 400 msec (P < .05). A subjective evaluation revealed an overall superiority of gray-matter differentiation with TI = 400 msec. CONCLUSION: A TI of 400 msec was the most suitable for this purpose.     

Pipeline flow diversion with adjunctive coil embolization for internal carotid artery aneurysms following an intradural component: results in 46 consecutive aneurysms from a Japanese single-center experience

In the treatment of an intracranial aneurysm with the flow diverter, the combined use of coil embolization can help promote subsequent progressive thrombosis within the aneurysm sac and reduce the risk of delayed aneurysm rupture. This study retrospectively reviewed outcomes of patients who had undergone the Pipeline Embolization Device (PED) with adjunctive coil embolization (PED/coil) at a single center to determine its safety and efficiency. Patients with internal carotid artery aneurysms following an intradural component were selected for PED/coil between 2015 and 2020. All patients were premedicated with dual antiplatelet therapy of aspirin plus clopidogrel or prasugrel. A minimal number of PEDs were deployed, with coils inserted using a stent-jail technique, avoiding dense packing. A total of 46 aneurysms (43 patients; median dome size, 11.6 mm; median neck width, 6.3 mm) were treated with PED/coil. The median volume embolization ratio was 14.8%. The degree of angiographic filling at the 6-month and latest angiography showed complete occlusion in 60.5% (26/43) and 70.5% (31/44), respectively. Small (<?10 mm) aneurysms achieved a higher complete occlusion rate in the early period; a lower cumulative incidence of aneurysm occlusion was observed in large and giant (≥?10 mm) aneurysms (P?=?.024). The median clinical follow-up was 22 months, and no aneurysm ruptures occurred. Favorable clinical outcomes were achieved, with permanent neurological morbidity of 4.7% and no mortality. PED/coil demonstrated a high angiographic occlusion rate at an early stage. Loosely packed coils are sufficient to obliterate aneurysms effectively.