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1.
Skeletal Radiol ; 53(5): 881-890, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-37935923

RESUMO

PURPOSE: To investigate the feasibility and application of a novel imaging technique, a three-dimensional dual adiabatic inversion recovery prepared ultrashort echo time (3D DIR-UTE) sequence, for high contrast assessment of cartilaginous endplate (CEP) imaging with head-to-head comparisons between other UTE imaging techniques. METHOD: The DIR-UTE sequence employs two narrow-band adiabatic full passage (AFP) pulses to suppress signals from long T2 water (e.g., nucleus pulposus (NP)) and bone marrow fat (BMF) independently, followed by multispoke UTE acquisition to detect signals from the CEP with short T2 relaxation times. The DIR-UTE sequence, in addition to three other UTE sequences namely, an IR-prepared and fat-saturated UTE (IR-FS-UTE), a T1-weighted and fat-saturated UTE sequence (T1w-FS-UTE), and a fat-saturated UTE (FS-UTE) was used for MR imaging on a 3 T scanner to image six asymptomatic volunteers, six patients with low back pain, as well as a human cadaveric specimen. The contrast-to-noise ratio of the CEP relative to the adjacent structures-specifically the NP and BMF-was then compared from the acquired images across the different UTE sequences. RESULTS: For asymptomatic volunteers, the DIR-UTE sequence showed significantly higher contrast-to-noise ratio values between the CEP and BMF (CNRCEP-BMF) (19.9 ± 3.0) and between the CEP and NP (CNRCEP-NP) (23.1 ± 1.7) compared to IR-FS-UTE (CNRCEP-BMF: 17.3 ± 1.2 and CNRCEP-NP: 19.1 ± 1.8), T1w-FS-UTE (CNRCEP-BMF: 9.0 ± 2.7 and CNRCEP-NP: 10.4 ± 3.5), and FS-UTE (CNRCEP-BMF: 7.7 ± 2.2 and CNRCEP-NP: 5.8 ± 2.4) for asymptomatic volunteers (all P-values < 0.001). For the spine sample and patients with low back pain, the DIR-UTE technique detected abnormalities such as irregularities and focal defects in the CEP regions. CONCLUSION: The 3D DIR-UTE sequence is able to provide high-contrast volumetric CEP imaging for human spines on a clinical 3 T scanner.


Assuntos
Dor Lombar , Humanos , Osso e Ossos , Imageamento por Ressonância Magnética/métodos , Cartilagem , Imagens de Fantasmas , Imageamento Tridimensional/métodos
2.
Gan To Kagaku Ryoho ; 50(11): 1207-1210, 2023 Nov.
Artigo em Japonês | MEDLINE | ID: mdl-38056876

RESUMO

A 79-year-old man visited a hospital for right upper abdominal pain and nausea. After conservative treatment for cholangitis and pancreatitis owing to a pancreatic head lesion, he was referred to our hospital for further evaluation and treatment of the lesion. He was diagnosed with pancreatic head cancer or carcinoma of papilla of Vater and underwent subtotal stomach- preserving pancreaticoduodenectomy. Postoperative histopathological examination revealed the coexistence of adenocarcinoma( 60%)and neuroendocrine carcinoma(40%)components, consistent with the diagnosis of mixed neuroendocrine- non-neuroendocrine neoplasm(MiNEN). In addition, regional lymph node metastasis of the adenocarcinoma component was found. Adjuvant chemotherapy was not administered because of a poor performance status. Lung metastasis occurred 13 months after surgery. Chemotherapy with S-1 was administered, and partial response was obtained 17 months after surgery. Herein, we report this rare case of MiNEN of the papilla of Vater with lung metastasis.


Assuntos
Adenocarcinoma , Ampola Hepatopancreática , Neoplasias Pulmonares , Neoplasias Pancreáticas , Masculino , Humanos , Idoso , Ampola Hepatopancreática/cirurgia , Ampola Hepatopancreática/patologia , Pancreaticoduodenectomia , Adenocarcinoma/cirurgia , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/cirurgia , Neoplasias Pulmonares/patologia , Neoplasias Pancreáticas/cirurgia , Pulmão/cirurgia
3.
Curr Eye Res ; 48(6): 564-572, 2023 06.
Artigo em Inglês | MEDLINE | ID: mdl-36852699

RESUMO

PURPOSE: Corneal limbal stem cell (LSC) transplantation has been reported as a potential approach to treat the damaged corneal epithelium. Scaffolds such as human amniotic membrane (hAM) are commonly employed for the in vitro culture and as a carrier during in vivo transplantation. However, they carry the risk of biological contamination and donor to donor variability. To overcome these disadvantages, we herein report the capabilities of a synthetic thermoreversible gelation polymer (TGP) scaffold to serve as an encapsulation support during LSC transplantation and to enable engraftment for corneal regeneration. METHODS: Sixteen discarded human corneas were used to isolate the corneal epithelium which was cultured in TGP and hAM. The cell proliferation and characteristics between TGP and hAM culture methods were evaluated by microscopic observation, 3H Thymidine incorporation assay, immunoperoxidase and immunofluorescence staining. RESULTS: The 3H Thymidine assay's results showed that TGP allowed human-donor cornea-derived LSCs to proliferate well in vitro, compared to hAM and the cells encapsulated in TGP and transplanted ex vivo onto a human cadaver donor cornea denuded of its epithelium, migrated on the ocular surface, and proliferated to form a continuous layer in 25 days. Immunoperoxidase and Immunofluorescence staining of TGP-cultured cells were positive for LSC markers (p63, ABCG2, Connexin 43 and Integrin ß), proving that the TGP helps to preserve the limbal cells' stemness. CONCLUSION: TGP is found to be a multipurpose scaffold for (i) in vitro culture, (ii) ex vivo encapsulation, and in vivo transplantation (iii), enabling engraftment of LSCs in this study, with potentials to extend its application in cell-based therapies in several regenerative medicine approaches.


Assuntos
Transplante de Córnea , Epitélio Corneano , Limbo da Córnea , Humanos , Córnea , Epitélio Corneano/metabolismo , Células Cultivadas , Proliferação de Células
4.
J Knee Surg ; 36(6): 667-672, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-34952550

RESUMO

Anterior cruciate ligament (ACL) injuries commonly lead to translational and rotational tibiofemoral instability. The morphology of the medial tibial eminence (MTE) has received increased attention regarding its role in tibiofemoral stability in ACL-injured knees. Therefore, quantification of MTE dimensions on clinical imaging may help clinicians predict knee stability after ACL injury. Although magnetic resonance imaging (MRI) is routinely obtained in patients with ACL injuries, whether the dimensions of the MTE can be accurate quantified on MRI is unknown. The purpose of this study was to assess the degree of correlation between measurements of MTE height and width on computed tomography (CT) versus MRI. An institutional picture archiving and communication system imaging database was used to identify patients aged between 15 and 60 years who received concurrent MRI and CT of the same knee within a 1-year interval. Knees with significant arthrosis, deformity, intraarticular fracture, or hardware-related artifact that obscured visualization of the MTE were excluded. Mean differences and interstudy agreement between CT and MRI MTE measurements were compared using concordance correlation coefficient (r c) and Bland-Altman analysis. A total of 41 knees in 38 patients (mean age, 37 years; 82% male) were analyzed. Interrater reliability for CT and MRI measurements was high (intraclass correlation coefficient = 0.740-0.954). On coronal CT and MRI, mean MTE height measurements were 10.4 ± 1.9 and 10.4 ± 1.8 mm, respectively; mean MTE width measurements were 14.6 ± 3.6 and 14.2 ± 3.0 mm, respectively. On sagittal CT and MRI, mean MTE height measurements were 11.6 ± 1.7 and 11.7 ± 1.7 mm, respectively; mean MTE width measurements were 36.5 ± 4.8 and 36.2 ± 5.0 mm, respectively. Good agreement was observed between CT and MRI measurements of MTE height and width on coronal and sagittal planes (r c = 0.947-0.969). Measurements of MTE height and width were similar on MRI relative to CT on both coronal and sagittal planes. MRI may be suitable for characterizing the dimensions of the MTE when clinically evaluating patients with ACL injuries, potentially allowing for individualized patient care.


Assuntos
Lesões do Ligamento Cruzado Anterior , Ligamento Cruzado Anterior , Humanos , Masculino , Adolescente , Adulto Jovem , Adulto , Pessoa de Meia-Idade , Feminino , Ligamento Cruzado Anterior/diagnóstico por imagem , Lesões do Ligamento Cruzado Anterior/diagnóstico por imagem , Lesões do Ligamento Cruzado Anterior/cirurgia , Reprodutibilidade dos Testes , Tíbia/diagnóstico por imagem , Tíbia/anatomia & histologia , Articulação do Joelho/diagnóstico por imagem , Articulação do Joelho/anatomia & histologia , Imageamento por Ressonância Magnética/métodos , Tomografia Computadorizada por Raios X
5.
Radiology ; 305(1): 160-166, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-35699577

RESUMO

Background Lumbar spine MRI studies are widely used for back pain assessment. Interpretation involves grading lumbar spinal stenosis, which is repetitive and time consuming. Deep learning (DL) could provide faster and more consistent interpretation. Purpose To assess the speed and interobserver agreement of radiologists for reporting lumbar spinal stenosis with and without DL assistance. Materials and Methods In this retrospective study, a DL model designed to assist radiologists in the interpretation of spinal canal, lateral recess, and neural foraminal stenoses on lumbar spine MRI scans was used. Randomly selected lumbar spine MRI studies obtained in patients with back pain who were 18 years and older over a 3-year period, from September 2015 to September 2018, were included in an internal test data set. Studies with instrumentation and scoliosis were excluded. Eight radiologists, each with 2-13 years of experience in spine MRI interpretation, reviewed studies with and without DL model assistance with a 1-month washout period. Time to diagnosis (in seconds) and interobserver agreement (using Gwet κ) were assessed for stenosis grading for each radiologist with and without the DL model and compared with test data set labels provided by an external musculoskeletal radiologist (with 32 years of experience) as the reference standard. Results Overall, 444 images in 25 patients (mean age, 51 years ± 20 [SD]; 14 women) were evaluated in a test data set. DL-assisted radiologists had a reduced interpretation time per spine MRI study, from a mean of 124-274 seconds (SD, 25-88 seconds) to 47-71 seconds (SD, 24-29 seconds) (P < .001). DL-assisted radiologists had either superior or equivalent interobserver agreement for all stenosis gradings compared with unassisted radiologists. DL-assisted general and in-training radiologists improved their interobserver agreement for four-class neural foraminal stenosis, with κ values of 0.71 and 0.70 (with DL) versus 0.39 and 0.39 (without DL), respectively (both P < .001). Conclusion Radiologists who were assisted by deep learning for interpretation of lumbar spinal stenosis on MRI scans showed a marked reduction in reporting time and superior or equivalent interobserver agreement for all stenosis gradings compared with radiologists who were unassisted by deep learning. © RSNA, 2022 Online supplemental material is available for this article. See also the editorial by Hayashi in this issue.


Assuntos
Aprendizado Profundo , Estenose Espinal , Constrição Patológica , Feminino , Humanos , Vértebras Lombares/diagnóstico por imagem , Imageamento por Ressonância Magnética/métodos , Pessoa de Meia-Idade , Estudos Retrospectivos , Canal Medular , Estenose Espinal/diagnóstico por imagem
6.
J Orthop ; 29: 60-64, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35145328

RESUMO

BACKGROUND: Chondrocytes are used in cell-based therapies such as autologous chondrocyte implantation (ACI) and matrix-associated cartilage implantation (MACI). To transport the cartilage tissue to the laboratory for in vitro culturing, phosphate-buffered saline (PBS), Euro-Collins solution (ECS) and Dulbecco's Modified Eagle's Medium (DMEM) are commonly employed at 4-8 °C. METHODS: In this study, eight samples of human cartilage biopsy tissues from elderly patients with severe osteoarthritis undergoing arthroscopy, which would otherwise have been discarded, were used. The cartilage tissue samples were compared to assess the cell yield between two transportation groups: i) a thermo-reversible gelation polymer (TGP) based method without cool preservation (∼25 °C) and ii) ECS transport at 4 °C. These samples were subjected to in vitro culture in a two-dimensional (2D) monolayer for two weeks and subsequently in a three-dimensional (3D) TGP scaffold for six weeks. RESULTS: The cell count obtained from the tissues transported in TGP was higher (0.2 million cells) than those transported in ECS (0.08 million cells) both after initial processing and after in vitro culturing for 2 weeks in 2D (18 million cells compared with 10 million cells). In addition, mRNA quantification demonstrated significantly higher expression of Col2a1 and SOX-9 in 3D-TGP cultured cells and lower expression of COL1a1 in RT-PCR, characteristic of the hyaline cartilage phenotype, than in 2D culture. CONCLUSION: This study confirms that the TGP cocktail is suitable for both the transport of human cartilage tissue and for in vitro culturing to yield better-quality cells for use in regenerative therapies.

7.
Magn Reson Med Sci ; 21(1): 29-40, 2022 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-34471014

RESUMO

Osteoarthritis (OA) is one of the most prevalent disorders in today's society, resulting in significant socio-economic costs and morbidity. MRI is widely used as a non-invasive imaging tool for OA of the knee. However, conventional knee MRI has limitations to detect subtle early cartilage degeneration before morphological changes are visually apparent. Novel MRI pulse sequences for cartilage assessment have recently received increased attention due to newly developed compositional MRI techniques, including: T2 mapping, T1rho mapping, delayed gadolinium-enhanced MRI of cartilage (dGEMRIC), sodium MRI, diffusion-weighted imaging (DWI)/ diffusion tensor imaging (DTI), ultrashort TE (uTE), and glycosaminoglycan specific chemical exchange saturation transfer (gagCEST) imaging. In this article, we will first review these quantitative assessments. Then, we will discuss the variations of quantitative values of knee articular cartilage with cartilage layer (depth)- and angle (regional)-dependent approaches. Multiple MRI sequence techniques can discern qualitative differences in knee cartilage. Normal articular hyaline cartilage has a zonal variation in T2 relaxation times with increasing T2 values from the subchondral bone to the articular surface. T1rho values were also higher in the superficial layer than in the deep layer in most locations in the medial and lateral femoral condyles, including the weight-bearing portion. Magic angle effect on T2 mapping is clearly observed in the both medial and lateral femoral condyles, especially within the deep layers. One of the limitations for clinical use of these compositional assessments is a long scan time. Recent new approaches with compressed sensing (CS) and MR fingerprinting (MRF) have potential to provide accurate and fast quantitative cartilage assessments.


Assuntos
Cartilagem Articular , Osteoartrite do Joelho , Cartilagem Articular/diagnóstico por imagem , Imagem de Tensor de Difusão , Humanos , Joelho , Articulação do Joelho/diagnóstico por imagem , Imageamento por Ressonância Magnética/métodos , Osteoartrite do Joelho/diagnóstico por imagem , Osteoartrite do Joelho/patologia
8.
Biopreserv Biobank ; 20(1): 97-103, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-34962137

RESUMO

Introduction: The transportation of tissues from hospitals to clinical laboratories for cell therapy is an essential component of regenerative medicine. Previously, we used laboratory-cultured mucosal cells from buccal epithelium expanded and encapsulated using a scaffold-hybrid approach to the urethral stricture (BEES-HAUS) procedure. In this study, to improve the outcomes, we compared the thermoreversible gelation polymer (TGP) transportation procedure with conventional culture methods, and reported its advantages. Methods: Human buccal mucosal tissues in Phase I of the study were transported in Euro-Collins solution (ECS) and the cells obtained were cultured in two-dimensional (2D) Dulbecco's modified Eagle's medium (DMEM), CnT-Prime epithelial 2D differentiation medium (CnT-PR), and a three-dimensional (3D)-TGP scaffold. In Phase II, tissues were transported in a TGP cocktail and the ECS. The cells were cultured in 2D-DMEM and 3D-TGP, quantified, and characterized by immunohistochemistry. Results: The cells in 3D-TGP culture maintained epithelial morphology in a better manner compared with 2D-DMEM, in which they developed fibroblast-like morphology. The TGP-transported cells grew rapidly. Immunohistochemical analysis results for AE1/AE3, EGFR, integrin-ß1, p63, and p75 were intensely positive in 3D-TGP. Conclusion: The TGP-based cocktail used in human buccal tissue transportation yielded cells with better morphology maintenance. The TGP scaffold provides an optimal in vitro environment wherein epithelial cells better maintain their native phenotype compared to those cultured through conventional methods. These results suggest using TGP for the transportation and culture of human buccal tissues for clinical applications. In addition, the use of a TGP-based cocktail for the transport of other tissues for regenerative medicine applications is worth further analysis.


Assuntos
Engenharia Tecidual , Estreitamento Uretral , Células Cultivadas , Células Epiteliais , Humanos , Polímeros , Engenharia Tecidual/métodos
9.
Sci Rep ; 11(1): 14059, 2021 07 07.
Artigo em Inglês | MEDLINE | ID: mdl-34234261

RESUMO

Regenerative medicine applications require cells that are not inflicted with senescence after in vitro culture for an optimal in vivo outcome. Methods to overcome replicative senescence include genomic modifications which have their own disadvantages. We have evaluated a three-dimensional (3D) thermo-reversible gelation polymer (TGP) matrix environment for its capabilities to reverse cellular senescence. The expression of senescence-associated beta-galactosidase (SA-ßgal) by human chondrocytes from osteoarthritis-affected cartilage tissue, grown in a conventional two-dimensional (2D) monolayer culture versus in 3D-TGP were compared. In 2D, the cells de-differentiated into fibroblasts, expressed higher SA-ßgal and started degenerating at 25 days. SA-ßgal levels decreased when the chondrocytes were transferred from the 2D to the 3D-TGP culture, with cells exhibiting a tissue-like growth until 42-45 days. Other senescence associated markers such as p16INK4a and p21 were also expressed only in 2D cultured cells but not in 3D-TGP tissue engineered cartilage. This is a first-of-its-kind report of a chemically synthesized and reproducible in vitro environment yielding an advantageous reversal of aging of human chondrocytes without any genomic modifications. The method is worth consideration as an optimal method for growing cells for regenerative medicine applications.


Assuntos
Biopolímeros , Senescência Celular/genética , Condrócitos/metabolismo , Regulação da Expressão Gênica , Engenharia Tecidual , Alicerces Teciduais , beta-Galactosidase/genética , Biomarcadores , Técnicas de Cultura de Células , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Citometria de Fluxo , Imunofluorescência , Humanos , Engenharia Tecidual/métodos , beta-Galactosidase/metabolismo
10.
J Biochem ; 170(4): 511-520, 2021 Dec 04.
Artigo em Inglês | MEDLINE | ID: mdl-34097066

RESUMO

Escherichia coli dihydropyrimidine dehydrogenase (EcDPD) catalyses the NADH-dependent reduction of uracil and thymine to the corresponding 5,6-dihydropyrimidines to control their metabolite pools. EcDPD consists of two subunits, PreT and PreA, and requires FAD, FMN and Fe-S clusters for activity. Recombinant EcDPD with a C-terminal His6-tagged-PreA subunit was overproduced in a DPD-lacking E. coli cells with augmented Fe-S cluster synthesis. Anaerobic purification resulted in purified enzyme with a specific activity of 13 µmol min-1 mg-1. The purified EcDPD was a heterotetramer and contained 0.81 FAD, 0.99 FMN, 14 acid-labile sulphur and 15 iron per PreT-PreA dimer. The enzyme exhibited Michaelis-Menten kinetics for both the forward and reverse reactions, which is distinct from mammalian DPDs showing substrate inhibition kinetics. For uracil reduction, the kcat, kcat/KNADH and kcat/Kuracil values were constant over the pH range of 5.5-10. For dihydrouracil (DHU) dehydrogenation, the pH-dependence of the kcat and kcat/KNAD+ values indicated that a residue with a pKa of 6.6 must be deprotonated for activity. Biochemical and kinetic comparisons with pig DPD revealed that protonation sates of the catalytically competent forms of EcDPD are distinct from those of pig enzyme.


Assuntos
Di-Hidrouracila Desidrogenase (NADP)/metabolismo , Proteínas de Escherichia coli/metabolismo , Flavoproteínas/metabolismo , Proteínas Ferro-Enxofre/metabolismo , Animais , Catálise , Escherichia coli/enzimologia , Escherichia coli/metabolismo , Proteínas de Escherichia coli/química , Flavinas/metabolismo , Flavoproteínas/química , Histidina/metabolismo , Ferro/química , Ferro/metabolismo , Cinética , Oligopeptídeos/metabolismo , Enxofre/química , Enxofre/metabolismo , Suínos , Timina/metabolismo , Uracila/análogos & derivados , Uracila/metabolismo
11.
Radiology ; 300(1): 130-138, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-33973835

RESUMO

Background Assessment of lumbar spinal stenosis at MRI is repetitive and time consuming. Deep learning (DL) could improve -productivity and the consistency of reporting. Purpose To develop a DL model for automated detection and classification of lumbar central canal, lateral recess, and neural -foraminal stenosis. Materials and Methods In this retrospective study, lumbar spine MRI scans obtained from September 2015 to September 2018 were included. Studies of patients with spinal instrumentation or studies with suboptimal image quality, as well as postgadolinium studies and studies of patients with scoliosis, were excluded. Axial T2-weighted and sagittal T1-weighted images were used. Studies were split into an internal training set (80%), validation set (9%), and test set (11%). Training data were labeled by four radiologists using predefined gradings (normal, mild, moderate, and severe). A two-component DL model was developed. First, a convolutional neural network (CNN) was trained to detect the region of interest (ROI), with a second CNN for classification. An internal test set was labeled by a musculoskeletal radiologist with 31 years of experience (reference standard) and two subspecialist radiologists (radiologist 1: A.M., 5 years of experience; radiologist 2: J.T.P.D.H., 9 years of experience). DL model performance on an external test set was evaluated. Detection recall (in percentage), interrater agreement (Gwet κ), sensitivity, and specificity were calculated. Results Overall, 446 MRI lumbar spine studies were analyzed (446 patients; mean age ± standard deviation, 52 years ± 19; 240 women), with 396 patients in the training (80%) and validation (9%) sets and 50 (11%) in the internal test set. For internal testing, DL model and radiologist central canal recall were greater than 99%, with reduced neural foramina recall for the DL model (84.5%) and radiologist 1 (83.9%) compared with radiologist 2 (97.1%) (P < .001). For internal testing, dichotomous classification (normal or mild vs moderate or severe) showed almost-perfect agreement for both radiologists and the DL model, with respective κ values of 0.98, 0.98, and 0.96 for the central canal; 0.92, 0.95, and 0.92 for lateral recesses; and 0.94, 0.95, and 0.89 for neural foramina (P < .001). External testing with 100 MRI scans of lumbar spines showed almost perfect agreement for the DL model for dichotomous classification of all ROIs (κ, 0.95-0.96; P < .001). Conclusion A deep learning model showed comparable agreement with subspecialist radiologists for detection and classification of central canal and lateral recess stenosis, with slightly lower agreement for neural foraminal stenosis at lumbar spine MRI. © RSNA, 2021 Online supplemental material is available for this article. See also the editorial by Hayashi in this issue.


Assuntos
Aprendizado Profundo , Interpretação de Imagem Assistida por Computador/métodos , Imageamento por Ressonância Magnética/métodos , Estenose Espinal/diagnóstico por imagem , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Vértebras Lombares/diagnóstico por imagem , Masculino , Pessoa de Meia-Idade , Reprodutibilidade dos Testes , Estudos Retrospectivos , Adulto Jovem
12.
Life Sci ; 278: 119553, 2021 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-33932445

RESUMO

AIMS: We have evaluated the potential of a three-dimensional (3D) thermoreversible gelation polymer (TGP) matrix in enhancing miRNA 140 expression (a biomarker correlating with homeostasis and cartilage regeneration) during the in vitro expansion of osteoarthritis (OA)-affected human chondrocytes. MATERIALS AND METHODS: OA-chondrocytes were cultured in two-dimensional (2D) monolayer followed by culture in 3D-TGP. miRNA 140 expression levels in cell culture supernatant followed by expression in the cell lysate of both 2D and 3D-TGP cultures were analyzed. KEY FINDINGS: The expression of miRNA 140 in cell culture supernatant from the 3D-TGP group was 0.001 to 0.002% that in 2D culture supernatant while in the cell lysate, miRNA 140 expression in the 3D-TGP was nearly 30-fold higher than that of 2D group. SIGNIFICANCE: The 3D-TGP matrix allows enhanced expression of miRNA 140 in OA-affected human chondrocytes in vitro which after necessary validations can be applied in clinical transplantation to significantly improve the outcome.


Assuntos
Condrócitos/patologia , MicroRNAs/genética , Osteoartrite/genética , Regulação para Cima , Técnicas de Cultura de Células , Células Cultivadas , Condrócitos/metabolismo , Humanos , Osteoartrite/patologia , Polímeros/química , Alicerces Teciduais/química
13.
IEEE J Biomed Health Inform ; 25(7): 2388-2397, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-34003757

RESUMO

We propose an interpretable and lightweight 3D deep neural network model that diagnoses anterior cruciate ligament (ACL) tears from a knee MRI exam. Previous works focused primarily on achieving better diagnostic accuracy but paid less attention to practical aspects such as explainability and model size. They mainly relied on ImageNet pre-trained 2D deep neural network backbones, such as AlexNet or ResNet, which are computationally expensive. Some of them tried to interpret the models using post-inference visualization tools, such as CAM or Grad-CAM, which lack in generating accurate heatmaps. Our work addresses the two limitations by understanding the characteristics of ACL tear diagnosis. We argue that the semantic features required for classifying ACL tears are locally confined and highly homogeneous. We harness the unique characteristics of the task by incorporating: 1) attention modules and Gaussian positional encoding to reinforce the seeking of local features; 2) squeeze modules and fewer convolutional filters to reflect the homogeneity of the features. As a result, our model is interpretable: our attention modules can precisely highlight the ACL region without any location information given to them. Our model is extremely lightweight: consisting of only 43 K trainable parameters and 7.1 G of Floating-point operations per second (FLOPs), that is 225 times smaller and 91 times lesser than the previous state-of-the-art, respectively. Our model is accurate: our model outperforms the previous state-of-the-art with the average ROC-AUC of 0.983 and 0.980 on the Chiba and Stanford knee datasets, respectively.


Assuntos
Lesões do Ligamento Cruzado Anterior , Aprendizado Profundo , Ligamento Cruzado Anterior , Lesões do Ligamento Cruzado Anterior/diagnóstico por imagem , Humanos , Imageamento por Ressonância Magnética , Redes Neurais de Computação
14.
Biotechnol Lett ; 43(6): 1241-1251, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33768381

RESUMO

PURPOSE: McCarey-Kaufman's (MK) medium and Optisol-GS medium are the most commonly employed media for human donor corneal preservation. In this study, we evaluated the preservation efficacy of discarded human donor corneas using a Thermo-reversible gelation polymer (TGP) added to these two media. METHODS: Thirteen human corneal buttons collected from deceased donors, which were otherwise discarded due to low endothelial cell density (ECD) were used. They were stored in four groups: MK medium, MK medium with TGP, Optisol-GS and Optisol-GS with TGP at 4 °C for 96 h. Slit lamp examination and specular microscopy were performed. Corneal limbal tissues from these corneas were then cultured using explant methodology one with and the other without TGP scaffold, for 21 days. RESULTS: MK + TGP and Optisol-GS + TGP preserved corneas better than without TGP, which was observed by maintenance of ECD which was significantly higher in Optisol-GS + TGP than MK + TGP (p-value = 0.000478) and corneal thickness remaining the same for 96 h. Viable corneal epithelial cells could be grown from the corneas stored only in MK + TGP and Optisol-GS + TGP. During culture, the TGP scaffold helped maintain the native epithelial phenotype and progenitor/stem cell growth was confirmed by RT-PCR characterization. CONCLUSION: TGP reconstituted with MK and Optisol-GS media yields better preservation of human corneal buttons in terms of relatively higher ECD maintenance and better in vitro culture outcome of corneal limbal tissue. This method has the potential to become a standard donor corneal transportation-preservation methodology and it can also be extended to other tissue or organ transportation upon further validation.


Assuntos
Meios de Cultura/química , Endotélio Corneano/citologia , Preservação de Tecido/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Cadáver , Sulfatos de Condroitina/química , Misturas Complexas/química , Dextranos/química , Feminino , Gentamicinas/química , Humanos , Masculino , Pessoa de Meia-Idade , Compostos Orgânicos/química , Microscopia com Lâmpada de Fenda
15.
J Dance Med Sci ; 25(1): 38-45, 2021 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-33706854

RESUMO

The ankles of ballet dancers are routinely under heavy loading that may lead to osteoarthritic changes. It would be clinically useful to identify such pathology as early as possible in a dancer's career. Therefore, the purpose of this study was to compare quantitative measurements in magnetic resonance (MR) images of the talocrural and talonavicular joints in ballet dancers and healthy non-dancers for use in formulating prediction of chronic injury and degenerative joint disease in these locations. Quantitative measurements in MR images of the talocrural and talonavicular joints were compared in 10 female ballet dancers, 10 healthy female non-dancers, and nine male ballet dancers. Fat-suppressed density-weighted proton, T1rho, and T2 mapping images were acquired with a 3.0 T MR scanner. Medial and lateral subchondral bone distance between the tibia and talus (MSBD and LSBD), axial navicular-talus axis angle (ANT angle), sagittal talar neck angle against the posterior talocalcaneal joint (TN angle), and curvature of navicular surface at the talonavicular joint were measured on sagittal images. The medial subchondral bone distance was found to be significantly larger in female dancers than female non-dancers (4.05 mm vs. 2.75 mm, p < 0.05), whereas there were no significant differences in LSBD (2.63 mm vs. 2.63 mm, p = 0.87). Axial navicular talus angles in female dancers were significantly larger than those in female non-dancers (38.9° vs. 24.3°, p < 0.05). There was a tendency for the TN angle to be smaller and navicular curvature (NC) to be larger in female dancers compared to female non-dancers, though the differences were not significant (TN angle: 16.6° vs. 22.3°, p = 0.09, and NC: 0.186 vs. 0.165, p = 0.28). There were no significant differences in T1rho or T2 values of talonavicular joint cartilage. These results show that the bony anatomy of dancers' ankles may adapt to the stresses placed on them by ballet.


Assuntos
Dança , Articulações Tarsianas , Feminino , Humanos , Imageamento por Ressonância Magnética , Masculino
16.
Knee ; 29: 365-373, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33690017

RESUMO

BACKGROUND: Chondrocyte transplantation to address cartilage damage is an established solution. Because hyaluronic acid (HA) is an essential component for homeostasis of the cartilage, in order to arrive at methodologies to utilize its advantages in cell-based therapies, we compared the HA retention capability of a thermoreversible gelation polymer scaffold-based environment (3D-TGP) with conventional in vitro cell culture methodologies. METHODS: Chondrocytes derived from osteoarthritis-affected knee joint cartilage of elderly patients were used and accomplished in three phases. In Phase I, the levels of HA secreted by chondrocytes were measured in culture supernatant. In Phase II, retention capacity of externally added HA was quantified indirectly by measuring the HA released in culture supernatant, and in Phase III, the expression of CD44 on cells was analysed by immunohistochemistry. RESULTS: In Phase I, the average HA in the 3D supernatant was 3% that of 2D. In phase II, 80% of externally added HA was detected in the 2D on day 7, while in 3D-TGP, only 0.1% was released until day 21. In Phase III, 2D yielded individual cells that started degenerating from the third week; in 3D-TGP cells grew for a longer duration, formed a tissue-like architecture with extracellular matrix with significantly intense staining of CD44 than 2D. CONCLUSION: The capability of the 3D-TGP culture environment to retain HA and support chondrocytes to grow with a tissue-like architecture expressing higher HA content is considered advantageous as it serves as an in vitro culture platform that enables tissue engineering of cartilage tissue with native hyaline phenotype and higher HA expression. The in vitro environment being conducive, based on this data, we also recommend that the TGP be tried as an encapsulation material in clinical studies of chondrocyte implantation for optimal clinical outcome.


Assuntos
Cartilagem Articular/citologia , Condrócitos/metabolismo , Ácido Hialurônico/metabolismo , Osteoartrite do Joelho/metabolismo , Alicerces Teciduais , Idoso , Células Cultivadas , Humanos , Receptores de Hialuronatos/metabolismo , Imuno-Histoquímica
17.
J Orthop ; 23: 138-141, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33510554

RESUMO

OBJECTIVE: We evaluated the expression of stem/progenitor biomarkers in osteoarthritic tissue derived chondrocytes cultured using a three-dimensional (3D) thermo-reversible gelation polymer (TGP). METHODS: The chondrocytes from discarded biopsy tissues obtained from human elderly patients with osteoarthritis were cultured using the 3D-TGP up to six weeks. RESULTS: The chondrocytes grew in a tissue-like manner, without de-differentiation into fibroblasts, and the cells thus tissue-engineered were proven positive for CD49e, OCT4, CD-105 and STRO-1 by immunohistochemistry. CONCLUSION: This study establishes the efficacy of this 3D-TGP platform for clinically useable in-vitro tissue-engineered cartilage for improvising the clinical outcome of cell therapy for cartilage repair.

18.
Gan To Kagaku Ryoho ; 48(1): 85-87, 2021 Jan.
Artigo em Japonês | MEDLINE | ID: mdl-33468730

RESUMO

A 53-year-old woman was admitted to our hospital because of hepatic dysfunction found during a medical checkup. Cholecystitis was suspected, and unenhanced computed tomography (CT) was initially performed because she had bronchial asthma. However, a tumor-like lesion was seen at the bottom of the gallbladder. Contrast-enhanced CT was performed 3 weeks later, and the tumor-like lesion was enhanced and had increased in size. Endoscopic ultrasound fine-needle aspiration did not reveal any signs of malignancy. Colonoscopy revealed ulcerations in the transverse colon, and invasion from gallbladder cancer was suspected. Our preoperative diagnosis was xanthogranulomatous cholecystitis, but gallbladder cancer could not be excluded. Gallbladder bed resection and partial resection of the transverse colon were performed. Intraoperative frozen section analysis did not reveal any malignant findings; hence, we considered that lymph node dissection was unnecessary. Pathological examination confirmed xanthogranulomatous cholecystitis with abscess formation. In cases of surgery for xanthogranulomatous cholecystitis, it is important to consider that this condition could coexist with gallbladder cancer.


Assuntos
Colecistite , Colo Transverso , Neoplasias da Vesícula Biliar , Colecistite/cirurgia , Feminino , Vesícula Biliar , Neoplasias da Vesícula Biliar/diagnóstico por imagem , Neoplasias da Vesícula Biliar/cirurgia , Granuloma/diagnóstico por imagem , Granuloma/cirurgia , Humanos , Pessoa de Meia-Idade , Xantomatose
19.
Eur J Radiol ; 134: 109418, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33302025

RESUMO

OBJECTIVES: Assessment of the ulnar attachment of the triangular fibrocartilage complex (TFCC) in a neutral forearm position remains challenging. Our study aims to evaluate the visibility of ulnar sided TFCC on 3 T MRI and compare isotropic 3D FSE sequences utilizing multiplanar reformation (MPR) with standard high-resolution 2D FSE sequences. METHODS: Ninety-nine MRI wrist studies in patients with wrist pain were retrospectively analyzed. Patients were scanned with a neutral forearm position and reviewed with isotropic 3D coronal FSE proton density-weighted images (PDWI) and 2D coronal FSE PDWI. MPR was used for 3D assessment. Visibility of the dorsal radioulnar ligament (DRUL), triangular ligament (TL), and volar radioulnar ligament (VRUL) was assessed by three raters utilizing a five-point grading scale. Grades were compared between 2D and 3D sequences. Intrarater and interrater reliability for the delineation of anatomic structures was measured by Spearman's rank correlation coefficient, Cohen's kappa, and percentage of exact agreement/agreement within a range of ±1 score point. RESULTS: Visibility grades in 3D were statistically significantly higher than those in 2D in all ligaments by all raters (p < 0.01). In Spearman's rank correlation coefficient and Cohen's kappa analysis, interrater correlations and agreements are variable but tended to be higher on 3D than on 2D. Both 2D and 3D sequences showed high intrarater exact agreement in all ligaments (80-91 % on 2D and 88-95 % on 3D). All exact interrater agreements on 3D were acceptable for TL (83-93 %) and acceptable to close to acceptable for VRUL (72-96 %). CONCLUSION: The utilization of isotopic 3D imaging combined with MPR function significantly improves visibility of ulnar attachment of the TFCC.


Assuntos
Fibrocartilagem Triangular , Humanos , Imageamento Tridimensional , Ligamentos , Imageamento por Ressonância Magnética , Reprodutibilidade dos Testes , Estudos Retrospectivos , Fibrocartilagem Triangular/diagnóstico por imagem
20.
Acta Radiol ; 61(10): 1365-1376, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32028774

RESUMO

BACKGROUND: Since ballet dancers begin their training before skeletal maturity, accurate and non-invasive identification of cartilage diseases is clinically important. Angle-dependent analysis of T1rho and T2 sequences can be useful for quantification of the composition of cartilage. PURPOSE: To investigate the angle-dependent T1rho and T2 profiles of ankle cartilage in non-dancers and dancers. MATERIAL AND METHODS: Ten female non-dancers, ten female dancers, and 9 male dancers were evaluated using T1rho and T2 mapping sequences. Manual segmentation of talar and tibial cartilage on these images was performed by two radiologists. Inter- and intra-rater reliabilities were calculated using intraclass correlation coefficients (ICCs) and Bland-Altman analysis. Mean thickness and volume of cartilage were estimated. Angle-dependent relaxation time profiles of talar and tibial cartilage were created. RESULTS: ICCs of the number of segmented pixels were poor to excellent. Bland-Altman plots indicated that differences were associated with segment sizes. Segmented cartilage on T1rho demonstrated larger thickness and volume than those on T2 in all populations. Male dancers showed larger cartilage thickness and volume than female dancers and non-dancers. Each cartilage demonstrated angular-dependent T1rho and T2 profiles. Minimal T1rho and T2 values were observed at approximately 180°-200°; higher values were seen at the angle closer to the magic angle. Minimal T2 value of talar cartilage of dancers was larger than that of non-dancers. CONCLUSION: In this small cohort study, regional and sex variations of ankle cartilage T1rho and T2 values in dancers and non-dancers were demonstrated using an angle-dependent approach.


Assuntos
Articulação do Tornozelo/diagnóstico por imagem , Doenças das Cartilagens/diagnóstico por imagem , Cartilagem Articular , Dança , Artropatias/diagnóstico por imagem , Imageamento por Ressonância Magnética/métodos , Adulto , Feminino , Humanos , Masculino
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