Transformed lung organoids have extensive applications in lung cancer modeling and drug screening. Traditional two-dimensional (2D) cultures fail to propagate a large subpopulation of murine primary tumors in vitro. However, three-dimensional (3D) air-liquid interface (ALI) cultures, which are employed to grow normal lung organoids, can be used to efficiently culture cancerous lung tumor cells. Here, we detail a procedure for cultivating genetically modified lung organoids in 3D-ALI cultures. This protocol contains two parts. The first part describes how to transduce lung epithelial cells, which are either freshly sorted from lungs or from actively growing murine organoids, with virus in order to modify gene expression. The target lung cells are incubated with virus for 1-2 h for transduction. Then, the transduced cells are thoroughly washed and mixed with stromal support cells and Matrigel and are loaded into transwell inserts for culture and validated for genetic modifications through downstream assays. The second part describes how to isolate tumor cells growing orthotopically in genetically engineered mouse models to produce organoid cell lines that can be used for ex vivo drug discovery assays. For this protocol, tumors are isolated from lungs of mice, finely chopped and washed. Then, tumor chunks are mixed with Matrigel for 3D-ALI culture. Finally, organoids budding from tumor chunks are trypsinized and passaged to establish an organoid line. Together these two protocols provide a promising platform to study the genesis, progression, and treatment of lung cancer.

OBJECTIVE: The aim of this study was to identify cone-beam computed tomography (CBCT) protocols that offer an optimal balance between effective dose (ED) and 3D model for orthognathic virtual surgery planning, using CT as a reference, and to assess whether such protocols can be defined based on technical image quality metrics.
METHODS: Eleven CBCT (VISO G7, Planmeca Oy, Helsinki, Finland) scan protocols were selected out of 32 candidate protocols, based on ED and technical image quality measurements. Next, an anthropomorphic RANDO SK150 phantom was scanned using these 11 CBCT protocols and 2 CT scanners for bone quantity assessments. The resulting DICOM (Digital Imaging and Communications in Medicine) files were converted into Standard Tessellation Language (STL) models that were used for bone volume and area measurements in the predefined orbital region to assess the validity of each CBCT protocol for virtual surgical planning.
RESULTS: The highest CBCT bone volume and area of the STL models were obtained using normal dose protocol (F2) and ultra-low dose protocol (J13), which resulted in 48% and 96% of the mean STL bone volume and 48% and 95% of the bone area measured on CT scanners, respectively.
CONCLUSIONS: The normal dose CBCT protocol \"F2\" offered optimal bone area and volume balance for STL. The optimal CBCT protocol can be defined using contrast-to-noise ratio and modulation transfer function values that were similar to those of the reference CT scanners\'. CBCT scanners with selected protocols can offer a viable alternative to CT scanners for acquiring STL models for virtual surgical planning at a lower effective dose.

BACKGROUND: Three-dimensional printing (3DP) is being integrated into surgical practice at a significant pace, from preprocedural planning to procedure simulation. 3DP is especially useful in surgical education, where printed models are highly accurate and customizable. The aim of this study was to evaluate how 3DP is being integrated most recently into surgical residency training.
METHODS: We performed a structured literature search of the OVID/MEDLINE, EMBASE, and PUBMED databases following the Preferred Reporting Items for Systematic Reviews and Meta-analyses guidelines. Articles published from 2016 to 2023 that met predefined inclusion and exclusion criteria were included. Data extracted included surgical subspecialty using 3DP, application of 3DP, and any reported satisfaction measures of trainees. A thorough analysis of pooled data was performed to evaluate satisfaction rates among studies.
RESULTS: A total of 85 studies were included. The median number of participants was 18 (interquartile range 10-27). Fourteen surgical disciplines were represented, with ear, nose, and throat/otolaryngology having the highest recorded utilization of 3DP models among residents and medical students (22.0%), followed by neurosurgery (14.0%) and urology (12.0%). 3DP models were created most frequently to model soft tissue (35.3%), bone (24.7%), vessel (14.1%), mixed (16.4%), or whole organs (6.66%) (Fig.1). Feedback from trainees was overwhelmingly positive regarding the fidelity of the models and their support for integration into their training programs. Among trainees, the combined satisfaction rate with their use in the curriculum was 95% (95% confidence interval, 0.92-0.97), and the satisfaction rate with the model fidelity was 90% (95% confidence interval, 0.86-0.94).
CONCLUSIONS: There is wide variation in the surgical specialties utilizing 3DP models in training. These models are effective in increasing trainee comfort with both common and rare scenarios and are associated with a high degree of resident support and satisfaction. Plastic surgery programs may benefit from the integration of this technology, potentially strengthening future surgical curricula. Objective evaluations of their pedagogic effects on residents are areas of future research.

BACKGROUND: Automated segmentation of 3-dimensional pulp space on cone-beam computed tomography images presents a significant opportunity for enhancing diagnosis, treatment planning, and clinical education in endodontics. The aim of this systematic review was to investigate the performance of artificial intelligence-driven automated pulp space segmentation on cone-beam computed tomography images.
METHODS: A comprehensive electronic search was performed using PubMed, Web of Science, and Cochrane databases, up until February 2024. Two independent reviewers participated in the selection of studies, data extraction, and evaluation of the included studies. Any disagreements were resolved by a third reviewer. The Quality Assessment of Diagnostic Accuracy Studies-2 tool was used to assess the risk of bias.
RESULTS: Thirteen studies that met the eligibility criteria were included. Most studies demonstrated high accuracy in their respective segmentation methods, although there was some variation across different structures (pulp chamber, root canal) and tooth types (single-rooted, multirooted). Automated segmentation showed slightly superior performance for segmenting the pulp chamber compared to the root canal and single-rooted teeth compared to multi-rooted ones. Furthermore, the second mesiobuccal (MB2) canalsegmentation also demonstrated high performance. In terms of time efficiency, the minimum time required for segmentation was 13 seconds.
CONCLUSIONS: Artificial intelligence-driven models demonstrated outstanding performance in pulp space segmentation. Nevertheless, these findings warrant careful interpretation, and their generalizability is limited due to the potential risk and low evidence level arising from inadequately detailed methodologies and inconsistent assessment techniques. In addition, there is room for further improvement, specifically for root canal segmentation and testing of artificial intelligence performance in artifact-induced images.

The increasing application of virtual surgical planning (VSP) in orthognathic surgery implies a critical need for accurate prediction of facial and skeletal shapes. The craniofacial relationship in patients with dentofacial deformities is still not understood, and transformations between facial and skeletal shapes remain a challenging task due to intricate anatomical structures and nonlinear relationships between the facial soft tissue and bones. In this study, a novel bidirectional 3-dimensional (3D) deep learning framework, named P2P-ConvGC, was developed and validated based on a large-scale data set for accurate subject-specific transformations between facial and skeletal shapes. Specifically, the 2-stage point-sampling strategy was used to generate multiple nonoverlapping point subsets to represent high-resolution facial and skeletal shapes. Facial and skeletal point subsets were separately input into the prediction system to predict the corresponding skeletal and facial point subsets via the skeletal prediction subnetwork and facial prediction subnetwork. For quantitative evaluation, the accuracy was calculated with shape errors and landmark errors between the predicted skeleton or face with corresponding ground truths. The shape error was calculated by comparing the predicted point sets with the ground truths, with P2P-ConvGC outperforming existing state-of-the-art algorithms including P2P-Net, P2P-ASNL, and P2P-Conv. The total landmark errors (Euclidean distances of craniomaxillofacial landmarks) of P2P-ConvGC in the upper skull, mandible, and facial soft tissues were 1.964 ± 0.904 mm, 2.398 ± 1.174 mm, and 2.226 ± 0.774 mm, respectively. Furthermore, the clinical feasibility of the bidirectional model was validated using a clinical cohort. The result demonstrated its prediction ability with average surface deviation errors of 0.895 ± 0.175 mm for facial prediction and 0.906 ± 0.082 mm for skeletal prediction. To conclude, our proposed model achieved good performance on the subject-specific prediction of facial and skeletal shapes and showed clinical application potential in postoperative facial prediction and VSP for orthognathic surgery.

BACKGROUND: Although the substrate in persistent atrial fibrillation (PeAF) is not limited to the pulmonary veins (PVs), PV isolation (PVI) remains the cornerstone ablation strategy.
OBJECTIVE: The aim of this study was to describe the mechanism of outgoing wavefronts (WFs) originating in the PV sleeves during PeAF.
METHODS: Eleven patients presenting for first-time PeAF ablation were recruited (mean age 63.1 ± 10.9 years, 91% men). A 64-electrode catheter (Constellation; 38 mm) was positioned within the PV under fluoroscopic guidance. An inverse mapping technique was used to reconstruct unipolar atrial electrograms on the PV surface, and the resulting phase maps were used to identify incoming and outgoing WFs at the PV junction and to classify focal and re-entrant activity within the PV sleeves.
RESULTS: During PeAF, the PVs gave rise to outgoing WFs with a frequency of 3.7 s-1 (Q1-Q3: 3.4-5.4 s-1) compared with 3.6 s-1 (Q1-Q3: 2.8-4.2 s-1) for incoming WFs. Circuitous macroscopic re-entry was the dominant mechanism driving outgoing WFs (frequency of re-entry 2.7 s-1 [Q1-Q3: 2.0-3.3 s-1] compared with focal activity 1.4 s-1 [Q1-Q3: 1.1-1.5 s-1]; P < 0.006). This was initiated by incoming WFs in 80% of cases. Consecutive focal activation from the same location was infrequent (10.0% ± 6.6%, n = 10). Rotors ≥360° were never observed. The median ratio (R) of outgoing to incoming WF frequency was 1.14 (Q1-Q3: 0.84-1.75), with R > 1 in 6 of 11 PVs.
CONCLUSIONS: Electric activity generated by PV sleeves during PeAF is due mainly to circuitous re-entry initiated by incoming waves, frequently with R > 1. That is, the PVs act less as drivers of atrial fibrillation than as \"echo chambers\" that sustain and amplify fibrillatory activity.

UNASSIGNED: Unintended secondary changes in the posterior tibial slope (PTS) and tibial torsion angle (TTA) may occur after medial open-wedge high tibial osteotomy (MOWHTO). In surgical procedures using patient-specific instruments (PSIs), it is essential to reproduce the PTS and TTA that were planned in simulations.
UNASSIGNED: To analyze the factors causing unintended sagittal and axial alignment changes after MOWHTO.
UNASSIGNED: Case series; Level of evidence, 4.
UNASSIGNED: Overall, 63 patients (70 knees) who underwent MOWHTO using a PSI between June 2020 and June 2023 were retrospectively reviewed. Preoperative and postoperative computed tomography scans were 3-dimensionally reconstructed. Simulated osteotomy was performed so that the weightbearing line could pass through the target point. A PSI gapper was 3-dimensionally printed to fit the posteromedial corner of the osteotomy gap in the simulated HTO model. After MOWHTO using the PSI gapper, the actual postoperative model was compared with the preoperative or simulation model. This assessment included PTS, TTA, hinge axis, and osteotomy-related parameters. Cortical breakage around the lateral hinge was evaluated to assess stability.
UNASSIGNED: The mean PTS and TTA did not change in the simulation. However, significant changes were observed in the actual postoperative PTS and TTA (change, -2.4°± 2.2° and -3.9°± 4.7°, respectively). The PTS was reduced, while the TTA decreased with internal rotation of the distal fragment. The difference in the axial hinge axis angle (AHA) between the simulation and actual surgery was the factor most correlated with the difference in the PTS (r = 0.625; P < .001). In regression analysis, the difference in the AHA was the only factor associated with the difference in the PTS (β = 0.558; P = .001), and there were no factors that showed any significant associations with the difference in the TTA. In subgroup analyses for the change in the TTA, the correction angle and anterior osteotomy angle were significantly higher in the more internal rotation group (P = .023 and P = .010, respectively). The TTA change was significantly higher in the unstable group with lateral cortical breakage (P = .018). The unstable group was more likely to show an internal rotation of ≥5° (odds ratio, 5.0; P = .007).
UNASSIGNED: The AHA was associated with a difference in the PTS between the simulation and actual surgery. The change in the TTA was caused by a combination of multiple factors, such as a large correction angle and anterior osteotomy angle, but mainly by instability of the lateral cortical hinge.

Glioblastoma (GBM) is the most aggressive adult primary brain tumor with nearly universal treatment resistance and recurrence. The mainstay of therapy remains maximal safe surgical resection followed by concurrent radiation therapy and temozolomide chemotherapy. Despite intensive investigation, alternative treatment options, such as immunotherapy or targeted molecular therapy, have yielded limited success to achieve long-term remission. This difficulty is partly due to the lack of pre-clinical models that fully recapitulate the intratumoral and intertumoral heterogeneity of GBM and the complex tumor microenvironment. Recently, GBM 3D organoids originating from resected patient tumors, genetic manipulation of induced pluripotent stem cell (iPSC)-derived brain organoids and bio-printing or fusion with non-malignant tissues have emerged as novel culture systems to portray the biology of GBM. Here, we highlight several methodologies for generating GBM organoids and discuss insights gained using such organoid models compared to classic modeling approaches using cell lines and xenografts. We also outline limitations of current GBM 3D organoids, most notably the difficulty retaining the tumor microenvironment, and discuss current efforts for improvements. Finally, we propose potential applications of organoid models for a deeper mechanistic understanding of GBM and therapeutic development.

Stent underexpansion in calcified coronary stenosis is an important predictor of major short- and long-term adverse cardiovascular events. In this case, we describe a novel method for assessing stent expansion using 3-dimensional stent reconstruction with C-arm motion compensated computed tomography.

UNASSIGNED: Assessment of 3-dimensional (3D) femoral head coverage is critical in evaluating, preoperative planning, and treating hip dysplasia.
UNASSIGNED: To (1) propose a mathematical model to establish 3D femoral head coverage using conventional computed tomography (CT), (2) determine the correlation of 2D parameters with 3D coverage, and (3) characterize the patterns of dysplasia based on 3D morphology.
UNASSIGNED: Cross-sectional study; Level of evidence, 3.
UNASSIGNED: We identified 30 patients (n = hips) with symptomatic dysplasia and 30 patients (n = hips) without dysplasia. Patients with dysplastic hips were matched with regard to sex, age, and body mass index to those with nondysplastic hips. Preoperative CTs were analyzed using 3D software, and 3D femoral head surface area coverage (FHSAC; in %) was assessed in 4 quadrant zones: anteromedial, anterolateral, posteromedial, and posterolateral. To assess lateral coverage of the femoral head, we introduced the anterolateral femoral head coverage angle (ALFC) and the posterolateral femoral head coverage angle (PLFC).
UNASSIGNED: Reduced femoral head coverage was more pronounced in dysplastic versus nondysplastic hips in the anterolateral quadrant (18% vs 40.7%, respectively) and posterolateral quadrant (35.8% vs 56.9%, respectively) (P < .0001 for both). Dysplastic hips had smaller ALFC and PLFC (18.4° vs 38.7°; P < .0001; 47.2° vs 72.3°; P = .0002). Anterolateral and posterolateral FHSAC were strongly correlated with the ALFC (r = 0.88; P < .0001) and the PLFC (r = 0.82; P < .0001) along with the lateral center-edge angle (anterolateral, r = 0.75; P < .0001; posterolateral, r = 0.73; P < .0001). Prediction models established for FHSAC had strong agreement with explanatory CT variables (anterolateral: r = 0.91; P < .0001; posterolateral: r = 0.90; P < .0001). The cutoff values for anterolateral and posterolateral FHSAC were 25% and 41%, respectively. In dysplastic hips, global deficiency was most common (15/30 hips), 9 hips showed an anterolateral deficiency, and 4 hips had a posterolateral deficiency pattern.
UNASSIGNED: The ALFC and The PLFC were strongly correlated with 3D lateral FHSAC and were able to predict 3D coverage accurately.