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1.
J Bone Joint Surg Am ; 2024 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-38743813

RESUMO

BACKGROUND: Ultrasonography is used to diagnose osteochondritis dissecans (OCD) of the humerus; however, its reliability depends on the technical proficiency of the examiner. Recently, computer-aided diagnosis (CAD) using deep learning has been applied in the field of medical science, and high diagnostic accuracy has been reported. We aimed to develop a deep learning-based CAD system for OCD detection on ultrasound images and to evaluate the accuracy of OCD detection using the CAD system. METHODS: The CAD process comprises 2 steps: humeral capitellum detection using an object-detection algorithm and OCD classification using an image classification network. Four-directional ultrasound images of the elbow of the throwing arm of 196 baseball players (mean age, 11.2 years), including 104 players with normal findings and 92 with OCD, were used for training and validation. An external dataset of 20 baseball players (10 with normal findings and 10 with OCD) was used to evaluate the accuracy of the CAD system. A confusion matrix and the area under the receiver operating characteristic curve (AUC) were used to evaluate the system. RESULTS: Clinical evaluation using the external dataset resulted in high AUCs in all 4 directions: 0.969 for the anterior long axis, 0.966 for the anterior short axis, 0.996 for the posterior long axis, and 0.993 for the posterior short axis. The accuracy of OCD detection thus exceeded 0.9 in all 4 directions. CONCLUSIONS: We propose a deep learning-based CAD system to detect OCD lesions on ultrasound images. The CAD system achieved high accuracy in all 4 directions of the elbow. This CAD system with a deep learning model may be useful for OCD screening during medical checkups to reduce the probability of missing an OCD lesion. LEVEL OF EVIDENCE: Diagnostic Level II. See Instructions for Authors for a complete description of levels of evidence.

2.
Artigo em Inglês | MEDLINE | ID: mdl-38233599

RESUMO

PURPOSE: Osteochondritis dissecans (OCD) of the humeral capitellum is a common cause of elbow disorders, particularly among young throwing athletes. Conservative treatment is the preferred treatment for managing OCD, and early intervention significantly influences the possibility of complete disease resolution. The purpose of this study is to develop a deep learning-based classification model in ultrasound images for computer-aided diagnosis. METHODS: This paper proposes a deep learning-based OCD classification method in ultrasound images. The proposed method first detects the humeral capitellum detection using YOLO and then estimates the OCD probability of the detected region probability using VGG16. We hypothesis that the performance will be improved by eliminating unnecessary regions. To validate the performance of the proposed method, it was applied to 158 subjects (OCD: 67, Normal: 91) using five-fold-cross-validation. RESULTS: The study demonstrated that the humeral capitellum detection achieved a mean average precision (mAP) of over 0.95, while OCD probability estimation achieved an average accuracy of 0.890, precision of 0.888, recall of 0.927, F1 score of 0.894, and an area under the curve (AUC) of 0.962. On the other hand, when the classification model was constructed for the entire image, accuracy, precision, recall, F1 score, and AUC were 0.806, 0.806, 0.932, 0.843, and 0.928, respectively. The findings suggest the high-performance potential of the proposed model for OCD classification in ultrasonic images. CONCLUSION: This paper introduces a deep learning-based OCD classification method. The experimental results emphasize the effectiveness of focusing on the humeral capitellum for OCD classification in ultrasound images. Future work should involve evaluating the effectiveness of employing the proposed method by physicians during medical check-ups for OCD.

3.
Biotechnol Lett ; 45(10): 1265-1277, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37606752

RESUMO

OBJECTIVES: Gene therapy using viral vectors and antibody-based therapies continue to expand within the pharmaceutical market. We evaluated whether Cellhesion® VP, a chitin-based material, can be used as 3D culture platform for cell lines used for the production of antibodies and viral vectors. RESULTS: The results of Cell Counting Kit-8 assay and LDH assay revealed that Cellhesion® VP had no adverse effect to Human Embryonic Kidney (HEK) 293, A549 and Chinese hamster ovary (CHO) DG44-Interferon-ß (IFN) cells. Cell growth analyses showed that Cellhesion® VP supported the 3D culture of HEK293, A549 and CHO DG44- IFN-ß cells with a spherical morphology. Importantly, subculture of these cell lines on Cellhesion® VP was easily performed without trypsinization because cells readily transferred to newly added scaffold. Our data also suggest that CHO DG44-IFNß, cultured on Cellhesion® VP secreted IFNß stably and continuously during the culture period. CONCLUSIONS: Cellhesion® VP provides a simple and streamlined expansion culture system for the production of biopharmaceuticals.


Assuntos
Produtos Biológicos , Animais , Cricetinae , Humanos , Células HEK293 , Quitina , Células CHO , Cricetulus , Técnicas de Cultura de Células
4.
J Orthop Surg Res ; 17(1): 568, 2022 Dec 27.
Artigo em Inglês | MEDLINE | ID: mdl-36575495

RESUMO

BACKGROUND: Diabetes mellitus weakens bone strength due to deterioration of bone quality; however, the histological mechanisms are still unknown. We hypothesized that histological assessment of cortical bone would enable us to determine the cause of the bone strength reduction associated with diabetes mellitus. Our aim was to evaluate the histomorphometric changes of cortical bone associated with deterioration of intrinsic bone properties and bone quality in diabetes mellitus. METHODS: We compared the outcomes of mechanical tests, bone mineral density measured using micro-computed tomography, and histological assessments, by applying Villanueva's bone stain, to the tibial bones of 40-week-old diabetic and control male rats. RESULTS: With respect to mechanical testing, the maximum load and energy absorption were significantly lower in the diabetic than in the control group, although fracture displacement and stiffness were not significantly different between the two groups. Bone mineral density was significantly higher in the diabetic group than in the control group. Bone histomorphometry revealed that the diabetic rats had fewer osteocytes, greater cortical porosity, and increased mineralization in cortical bone compared with the control group. CONCLUSIONS: Increased mineralization of the cortical bone with greater cortical porosity leads to a weakening of bone strength in diabetes mellitus.


Assuntos
Diabetes Mellitus Experimental , Ratos , Masculino , Animais , Diabetes Mellitus Experimental/complicações , Microtomografia por Raio-X/métodos , Osso e Ossos/diagnóstico por imagem , Densidade Óssea , Osso Cortical/diagnóstico por imagem
5.
BMC Musculoskelet Disord ; 23(1): 16, 2022 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-34980094

RESUMO

BACKGROUND: Teriparatide (TPTD) is a drug for osteoporosis that promotes bone formation and improves bone quality. However, the effects of TPTD on cortical bone are not well understood. Sweep imaging with Fourier transform (SWIFT) has been reported as a useful tool for evaluating bound water of cortical bone, but it has yet to be used to investigate the effects of TPTD on cortical bone. This study aimed to evaluate the consequences of the effect of TPTD on cortical bone formation using SWIFT. METHODS: Twelve-week-old female Sprague-Dawley rats (n = 36) were reared after ovariectomy to create a postmenopausal osteoporosis model. They were divided into two groups: the TPTD and non-TPTD groups. Rats were euthanized at 4, 12, and 24 weeks after initiating TPTD treatment. Tibial bones were evaluated using magnetic resonance imaging (MRI) and bone histomorphometry. In MRI, proton density-weighted imaging (PDWI) and SWIFT imaging were performed. The signal-to-noise ratio (SNR) was calculated for each method. The same area evaluated by MRI was then used to calculate the bone formation rate by bone histomorphometry. Measurements were compared using the Mann-Whitney U-test, and a P-value of < 0.05 was considered significant. RESULTS: PDWI-SNR was not significantly different between the two groups at any time point (P = 0.589, 0.394, and 0.394 at 4, 12, and 24 weeks, respectively). Contrarily, SWIFT-SNR was significantly higher in the TPTD group than in the non-TPTD group at 4 weeks after initiating treatment, but it was not significantly different at 12 and 24 weeks (P = 0.009, 0.937, and 0.818 at 4, 12, and 24 weeks, respectively). The bone formation rate assessed by histomorphometry was significantly higher in the TPTD group than in the non-TPTD group at all timepoints (P < 0.05, all weeks). In particular, at 4 weeks, the bone formation rate was markedly higher in the TPTD group than in the non-TPTD group (P = 0.028, 1.98 ± 0.33 vs. 0.09 ± 0.05 µm3/µm2/day). CONCLUSIONS: SWIFT could detect increased signals of bound water, reflecting the effect of TPTD on the cortical bone. The signal detected by SWIFT reflects a marked increase in the cortical bone formation rate.


Assuntos
Conservadores da Densidade Óssea , Teriparatida , Animais , Densidade Óssea , Conservadores da Densidade Óssea/farmacologia , Conservadores da Densidade Óssea/uso terapêutico , Osso Cortical/diagnóstico por imagem , Feminino , Análise de Fourier , Humanos , Imageamento por Ressonância Magnética , Osteogênese , Ratos , Ratos Sprague-Dawley , Teriparatida/farmacologia , Teriparatida/uso terapêutico
6.
Commun Biol ; 4(1): 1316, 2021 11 19.
Artigo em Inglês | MEDLINE | ID: mdl-34799690

RESUMO

Three-dimensional aggregate-suspension culture is a potential biomanufacturing method to produce a large number of human induced pluripotent stem cells (hiPSCs); however, the use of expensive growth factors and method-induced mechanical stress potentially result in inefficient production costs and difficulties in preserving pluripotency, respectively. Here, we developed a simple, miniaturized, dual-compartment dialysis-culture device based on a conventional membrane-culture insert with deep well plates. The device improved cell expansion up to approximately ~3.2 to 4×107 cells/mL. The high-density expansion was supported by reduction of excessive shear stress and agglomeration mediated by the addition of the functional polymer FP003. The results revealed accumulation of several growth factors, including fibroblast growth factor 2 and insulin, along with endogenous Nodal, which acts as a substitute for depleted transforming growth factor-ß1 in maintaining pluripotency. Because we used the same growth-factor formulation per volume in the upper culture compartment, the cost reduced in inverse proportional manner with the cell density. We showed that growth-factor-accumulation dynamics in a low-shear-stress environment successfully improved hiPSC proliferation, pluripotency, and differentiation potential. This miniaturised dialysis-culture system demonstrated the feasibility of cost-effective mass production of hiPSCs in high-density culture.


Assuntos
Técnicas de Cultura de Células/instrumentação , Diferenciação Celular , Células-Tronco Pluripotentes Induzidas/fisiologia , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/administração & dosagem
7.
JBJS Case Connect ; 10(4): e20.00279, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33749226

RESUMO

CASE: We describe a case of cryptococcal osteomyelitis in a 70-year-old man who presented with left elbow and shoulder pain. Subsequent imaging indicated osteomyelitis of the entire humerus, and he underwent debridement of the elbow and shoulder. Pathological findings revealed Cryptococcus neoformans infection. Surgical management was followed by a prolonged course of antibiotics. CONCLUSION: Although C. neoformans is rare as the etiology of infection of the entire humerus, orthopaedic surgeons should consider cryptococcosis as a potential cause of infection.


Assuntos
Criptococose/microbiologia , Cryptococcus neoformans/isolamento & purificação , Úmero/microbiologia , Osteomielite/microbiologia , Idoso , Criptococose/diagnóstico por imagem , Criptococose/imunologia , Humanos , Úmero/diagnóstico por imagem , Hospedeiro Imunocomprometido , Imageamento por Ressonância Magnética , Masculino , Osteomielite/diagnóstico por imagem
8.
PLoS One ; 13(9): e0200515, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30240390

RESUMO

Scalable production of avian cell lines exhibits a valuable potential on therapeutic application by producing recombinant proteins and as the substrate for virus growth due to the special glycosylation occurs in avian species. Chicken primordial germ cells (cPGCs), a germinal pluripotent avian cell type, present the ability of self-renewal, an anchorage-independent cell growth and the ability to be genetically modified. This cell type could be an interesting bioreactor system for industrial purposes. This study sought to establish an expandable culture system with defined components for three-dimensional (3D) culture of cPGCs. cPGCs were cultured in medium supplemented with the functional polymer FP003. Viscoelasticity was low in this medium but cPGCs did not sediment in culture and efficiencies of space and nutrient utilization were thus enhanced and consequently their expansion was improved. The total number of cPGCs increased by 17-fold after 1 week of culture in 3D-FAot medium, an aseric defined medium containing FP003 polymer, FGF2 and Activin A as growth factors and Ovotransferrin as protein. Moreover, cPGC cell lines stably expressed the germline-specific reporter VASA:tdTOMATO, as well as other markers of cPGCs, for more than 1 month upon culture in 3D-FAot medium, indicating that the characteristics of these cells are maintained. In summary, this novel 3D culture system can be used to efficiently expand cPGCs in suspension without mechanical stirring, which is available for long-term culture and no loss of cellular properties was found. This system provides a platform for large-scale culture of cPGCs.


Assuntos
Técnicas de Cultura de Células/métodos , Meios de Cultura/química , Meios de Cultura/farmacologia , Células Germinativas/citologia , Células Germinativas/metabolismo , Animais , Embrião de Galinha , Galinhas
9.
J Magn Reson Imaging ; 48(2): 389-397, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-29360263

RESUMO

BACKGROUND: Diabetes decreases bone strength, possibly because of cortical bone changes. Sweep imaging with Fourier transform (SWIFT) has been reported to be useful for cortical bone evaluation. PURPOSE: To evaluate cortical bone changes in diabetic rats using SWIFT, assess the usefulness of this technique through comparisons with microcomputed tomography (µCT) and conventional MRI, and clarify the mechanism underlying cortical bone changes using histomorphometry STUDY TYPE: Animal cohort. ANIMAL MODEL: 8-week-old male Wistar/ST rats (N = 36) were divided into diabetes (induced by streptozotocin injection) and control groups. FIELD STRENGTH/SEQUENCE: 7.04T MRI, SWIFT. ASSESSMENT: Six animals from each group were sacrificed at 2, 4, and 8 weeks after injection. Tibial bones were extracted and evaluated using µCT and MRI. The cortical bone mineral density (BMD) was measured using µCT. Proton density-weighted imaging (PDWI) and SWIFT were also performed. The signal-to-noise ratio (SNR) was calculated for each acquisition. The bone formation rate was evaluated using histomorphometry. STATISTICAL TESTS: Findings at each timepoint were compared using Mann-Whitney U-tests. RESULTS: Cortical BMD was significantly lower in the diabetes group than in the control group only at 8 weeks (P < 0.05). At all timepoints, PDWI-SNR showed no significant differences between groups (P = 0.59, 0.70, and 0.82 at 2, 4, and 8 weeks, respectively). SWIFT-SNR was significantly lower in the diabetes group than in the control group (P < 0.05 at 2 and 4 weeks and P < 0.01 at 8 weeks), and the bone formation rate was significantly lower in the diabetes group than in the control group (P < 0.01 for all). DATA CONCLUSION: SWIFT can detect cortical bone changes even before a decline in the cortical BMD in a diabetic model. LEVEL OF EVIDENCE: 1 Technical Efficacy: Stage 3 J. MAGN. RESON. IMAGING 2018;48:389-397.


Assuntos
Osso Cortical/diagnóstico por imagem , Diabetes Mellitus Experimental/diagnóstico por imagem , Análise de Fourier , Imageamento por Ressonância Magnética , Animais , Glicemia , Densidade Óssea , Masculino , Variações Dependentes do Observador , Ratos , Ratos Wistar , Razão Sinal-Ruído , Estresse Mecânico , Microtomografia por Raio-X
10.
Org Biomol Chem ; 7(18): 3772-81, 2009 Sep 21.
Artigo em Inglês | MEDLINE | ID: mdl-19707682

RESUMO

The total syntheses of (-)-cylindrocyclophane A (1), a naturally occurring, cytotoxic [7.7]paracyclophane, and its enantiomer have been achieved in an enantiodivergent manner starting from a chiral propargyl alcohol building block using Smith's cross metathesis/ring-closing metathesis protocol as the key step. The biological evaluation of both enantiomers of cylindrocyclophane A (1 and ent-1) and its analogues indicated that the chirality of 1 is irrelevant to its cytotoxicity, which is attributed to the resorcinol motifs embedded in the robust [7.7]paracyclophane framework.


Assuntos
Produtos Biológicos/síntese química , Produtos Biológicos/farmacologia , Hidrocarbonetos Policíclicos Aromáticos/síntese química , Hidrocarbonetos Policíclicos Aromáticos/farmacologia , Compostos Policíclicos/síntese química , Compostos Policíclicos/farmacologia , Álcoois/química , Antineoplásicos/síntese química , Antineoplásicos/química , Antineoplásicos/farmacologia , Produtos Biológicos/química , Proliferação de Células , Células HCT116 , Humanos , Hidrocarbonetos Policíclicos Aromáticos/química , Compostos Policíclicos/química , Estereoisomerismo , Relação Estrutura-Atividade
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