Minimum 10-year Survivorship of Uncemented Acetabular Reconstruction with Bulk Femoral Head Autografting in Crowe Type IV Developmental Hip Dysplasia.

BACKGROUND: In total hip arthroplasty (THA) for severe dislocations such as Crowe type IV developmental dysplasia of the hip (DDH), sufficient bone volume for stable fixation of the acetabular component can be achieved by placing a reinforcing bone graft prepared from the resected femoral head into the deficient acetabulum. The purpose of the current study was to examine the long-term survivorship of uncemented THA in conjunction with a bulk femoral head autograft in patients who have Crowe type IV DDH. METHODS: A total of 35 patients (42 hips) who have Crowe type IV DDH and underwent THA using uncemented cup fixation with bulk femoral head autografting were followed up for a mean period of 15.0 years (range, 10.0 to 20.0) postoperatively. Anteroposterior pelvic radiographs were used for measurements such as the horizontal coverage of the grafted bone and the center-edge angle. Kaplan-Meier survivorship analyses were performed with revision of the acetabular component as the endpoint. RESULTS: The Kaplan-Meier analysis indicated 15-year survival rates of 90.4%. The mean horizontal coverage of grafted bone was 46.1% (range, 23.7 to 66.0), and there were 16 cases with horizontal coverage of ≥ 50%. There was no difference in the appearance of a thin (< 1 mm) radiolucency line around the cup between cases with < 50% versus ≥ 50% of the horizontal coverage of grafted bone (4 versus 2 hips; P = 0.446). Trabecular bridging and remodeling were seen in all cases after mean periods of 4.1 months and 9.0 months postoperatively, respectively. Trabecular reorientation was seen in 41 of 42 hips (97.6%) at a mean follow-up of 19.9 months. CONCLUSIONS: Acetabular reconstruction with femoral bulk bone grafting for Crowe type IV DDH resulted in high survival rates and was a good method to restore bone stock and obtain long-term fixation.

Bone union-promoting effect of romosozumab in an ovariectomized rat posterolateral lumbar fusion model.

Spinal fixation surgery has been increasingly performed in patients with osteoporosis. Romosozumab, a drug that was introduced in Japan recently, is known to possibly promote bone healing. However, few studies have reported the therapeutic effects of romosozumab in clinical practice in Japan. Therefore, here, we investigated the effects of romosozumab dosage on bone fusion promotion using an ovariectomized rat spinal fusion model. Eight-week-old female Sprague-Dawley rats were matched by body weight and divided into three groups: 1.0 romosozumab (R) group (Evenity®, 25 mg/kg), 1/10R group (Evenity®, 2.5 mg/kg), and control (C) group (saline). Subcutaneous injections were administered twice a week for 8 weeks postoperatively. Computed tomography scans were performed every 2 weeks from the time of surgery till 8 weeks postoperatively. The mean fusion rates in terms of volume were significantly higher in the R groups [1/10R, 1.0R] than in the C group from 4 weeks postoperatively. The rate of increase was significantly higher in the 1.0R group from 4 weeks postoperatively and in the 1/10R group from 6 weeks postoperatively, than in the C group. The proportion of trabecular bone area was approximately 1.5 times higher in the R groups than in the C group. No significant differences were observed between the R groups. Our results suggest that romosozumab stimulates bone growth at the graft site, and similar effects were achieved at 1/10 of the standard dosage.

A Novel Evaluation for Vertebral Artery Course Using 3D Magnetic Resonance Imaging with Computed Tomography -like Bone Contrast and Magnetic Resonance Angiography: A Proof of Concept Study.

OBJECTIVE: Vertebral artery (VA) injury poses a significant risk in cervical spine surgery, necessitating accurate preoperative assessment. This study aims to introduce and validate a novel approach that combines the Fast field echo that resembles a computed tomography using restricted echo spacing (FRACTURE) sequence with Time of Flight (TOF) Magnetic Resonance Angiography (MRA) for comprehensive evaluation of VA courses in the cervical spine. MATERIALS AND METHODS: A total of eight healthy volunteers and two patients participated in this study. The FRACTURE sequence provided high-resolution bone images of the cervical spine, while TOF MRA offered non-invasive vascular imaging. Fusion images were created by merging FRACTURE and MRA modalities to simultaneously visualize cervical spine structures and VA courses. Board-certified orthopedic spine surgeons independently evaluated images to assess the visibility of anatomical characteristics of the VA course by Likert-scale. RESULTS: The FRACTURE-MRA fusion images effectively depicted the extraosseous course of the VA at the craniovertebral junction, the intraosseous course of the VA at the craniovertebral junction, the VA entrance level to the transverse foramen, and the side-to-side asymmetry of bilateral VAs. Additionally, clinical cases demonstrated the utility of the proposed technique in identifying anomalies and guiding surgical interventions. CONCLUSIONS: The integration of the FRACTURE sequence and TOF MRA presents a promising methodology for the precise evaluation of VA courses in the cervical spine. This approach improves preoperative planning for cervical spine surgery with detailed anatomy and is a valuable alternative to conventional methods without contrast agents.

The γ-glutamyl cycle serves as an amino acids supply system in colorectal cancer organoids under chronic hypoxia.

Malignant tumors are characterized by a hypoxic microenvironment, and metabolic reprogramming is necessary to ensure energy production and oxidative stress resistance. Although the microenvironmental properties of tumors vary under acute and chronic hypoxia, studies on chronic hypoxia-induced metabolic changes are limited. In the present study, we performed a comprehensive metabolic analysis in a chronic hypoxia model using colorectal cancer (CRC) organoids, and identified an amino acid supply system through the γ-glutamyl cycle, a glutathione recycling pathway. We analyzed the metabolic changes caused by hypoxia over time and observed that chronic hypoxia resulted in an increase in 5-oxoproline and a decrease in oxidized glutathione (GSSG) compared to acute hypoxia. These findings suggest that chronic hypoxia induces metabolic changes in the γ-glutamyl cycle. Moreover, inhibition of the γ-glutamyl cycle via γ-glutamyl cyclotransferase (GGCT) and γ-glutamyl transferase 1 (GGT1) knockdown significantly reversed chronic hypoxia-induced upregulation of 5-oxoproline and several amino acids. Notably, GGT1 knockdown downregulated the intracellular levels of γ-glutamyl amino acids. Conclusively, these results indicate that the γ-glutamyl cycle serves as an amino acid supply system in CRC under chronic hypoxia, which provides fresh insight into cancer metabolism under chronic hypoxia.

Machine learning algorithms for predicting Cobb angle beyond 25 degrees in female adolescent idiopathic scoliosis patients.

STUDY DESIGN: Retrospective cohort study. OBJECTIVE: To develop a machine learning (ML) model that predicts the progression of AIS using minimal radiographs and simple questionnaires during the first visit. SUMMARY OF BACKGROUND DATA: Several factors are associated with angle progression in patients with AIS. However, it is challenging to predict angular progression at the first visit. METHODS: Among female patients with AIS treated at a single institution from July 2011 to February 2023, 1119 cases were studied. Patient data, including demographic and radiographic data based on anterior-posterior and lateral whole-spine radiographs, were collected at the first and last visits. The last visit was defined differently based on treatment plans. For patients slated for surgery or bracing, the last visit occurred just before these interventions. For others, it was their final visit before turning 18 years. Angular progression was defined as a Cobb angle greater than 25 degrees for each of the proximal thoracic (PT), main thoracic (MT), and thoracolumbar/lumbar (TLL) curves at the last visit. ML algorithms were employed to develop individual binary classification models for each type of curve (PT, MT, and TLL) using PyCaret in Python. Multiple models were explored and analyzed, with the selection of optimal models based on the area under the curve (AUC) and Recall scores. Feature importance was evaluated to understand the contribution of each feature to the model predictions. RESULTS: For PT, MT, and TLL progression, the top-performing models exhibit AUC values of 0.94, 0.89, and 0.84, and achieve recall rates of 0.90, 0.85, and 0.81. The most significant factors predicting progression varied for each curve: initial Cobb angle for PT, presence of menarche for MT, and Risser grade for TLL. CONCLUSIONS: This study introduces an ML-based model using simple data at the first visit to precisely predict angle progression in female patients with AIS.

Machine Learning and Deep Learning in Spinal Injury: A Narrative Review of Algorithms in Diagnosis and Prognosis.

Spinal injuries, including cervical and thoracolumbar fractures, continue to be a major public health concern. Recent advancements in machine learning and deep learning technologies offer exciting prospects for improving both diagnostic and prognostic approaches in spinal injury care. This narrative review systematically explores the practical utility of these computational methods, with a focus on their application in imaging techniques such as computed tomography (CT) and magnetic resonance imaging (MRI), as well as in structured clinical data. Of the 39 studies included, 34 were focused on diagnostic applications, chiefly using deep learning to carry out tasks like vertebral fracture identification, differentiation between benign and malignant fractures, and AO fracture classification. The remaining five were prognostic, using machine learning to analyze parameters for predicting outcomes such as vertebral collapse and future fracture risk. This review highlights the potential benefit of machine learning and deep learning in spinal injury care, especially their roles in enhancing diagnostic capabilities, detailed fracture characterization, risk assessments, and individualized treatment planning.

[A Case of Testicular Torsion of Retractile Testis Requiring Orchidectomy].

We report a case of testicular torsion in an 8-year-old who was referred to our hospital for right groin pain. He was diagnosed with right retractile testis during a 12-month check-up. However, instead of performing orchiopexy, he was placed under observation until the age of 5, after which he did not seek medical attention. Physical examination revealed swelling and tenderness in the right inguinal region and no palpable testis in the right scrotum. Ultrasound and computed tomography revealed right testicular torsion, and emergency surgery was performed. Intraoperative findings revealed a dark and ischemic testis that was twisted at 180°in the right inguinal region. There was no improvement in blood flow even after the testicular torsion was released; therefore, right orchidectomy with left orchiopexy was performed. Although the incidence of testicular torsion is higher in patients with an undescended testis than in those with a normally positioned scrotal position testis, reports of testicular torsion associated with a retractile testis are rare.

Quantitative evaluation of the lumbar ligamentum flavum using MRI T2-mapping: Efficacy of its clinical application in patients with lumbar spinal stenosis.

OBEJECTIVE: To perform a magnetic resonance imaging T2-mapping of the ligamentum flavum in healthy individuals and patients with lumbar spinal stenosis scheduled for surgery and compare the T2 relaxation times. SUBJECTS AND METHODS: The T2 relaxation time of the ligamentum flavum was compared among 3 groups, healthy young individuals (H group (age< 50)), healthy middle-aged and older individuals (H group (age≥50)), and patients with lumbar spinal stenosis (L group). Additionally, the thickness of the ligament was measured in the axial image plane, and the occupied area ratio of each fiber was measured by staining the surgically obtained ligament, and each was correlated with the T2 relaxation time. We also evaluated the adhesion of the ligamentum flavum with the dura mater during the surgery. RESULTS: The T2 relaxation times were significantly prolonged in H group (age ≥50) and L group (P < 0.001) compared to H group (age<50). The relationship between collagen fiber and T2 relaxation times was significantly positive (r = 0.720, P < 0.001). Moreover, the relaxation times were significantly prolonged in those with adhesion of the ligamentum flavum with the dura mater (P < 0.05). The cut-off for the relaxation time was 50 ms (sensitivity: 62.50%, false positive rate: 10.8%). CONCLUSION: Healthy middle-aged and older individuals and patients with lumbar spinal stenosis and adhesion of the ligamentum flavum with the dura mater have prolonged T2 relaxation times. Hence, the adhesion between the ligamentum flavum and dura mater should be considered in cases with a relaxation time ≥50 ms.

Does vacuum phenomenon at non-fused discs affect the postoperative course after transforaminal lumbar interbody fusion in patients showing a positive value of difference in lumbar lordosis?

INTRODUCTION: Preoperative difference in lumbar lordosis (DiLL) was associated with surgical outcomes after single-level transforaminal lumbar interbody fusion (TLIF). Patients with DiLL>0 (DiLL (+)) tended to show worse clinical outcomes and postoperative greater restoration of lumbar lordosis (LL). However, some patients with DiLL (+) showed relatively good outcomes and no postoperative LL restration. This study aimed to elucidate whether the lumbar intervertebral disc vacuum phenomenon (VP) influences clinical course after single-level TLIF in patients with DiLL (+) and DiLL (-). METHODS: Patients with lumbar spinal stenosis and degenerative spondylolisthesis treated with single-level TLIF were included. Pre- and postoperative LL were measured, and postoperative LL improvement was calculated. Preoperative DiLL was calculated as preoperative supine LL minus standing LL. Severity of VP at the non-fused discs (SVP (non-FS)) was evaluated using preoperative reconstructed computed tomography imaging. Clinical outcomes were assessed using the Oswestry disability index, visual analogue scale (VAS; low back pain (LBP), lower-extremity pain, numbness, and the Japanese Orthopaedic Association Back Pain Evaluation Questionnaire. Patients were stratified by the median preoperative SVP (non-FS) score into severe and mild VP groups in patients with DiLL (+) or DiLL (-), and their surgical outcomes were compared. RESULTS: Overall, 89 patients were included. In patients with DiLL (+) (n = 37), patients with severe VP showed worse clinical outcomes, particulary for LBP and DiLL (+) patients with mild VP showed greater LL improvement (6.5° ± 10.0°). In patients with DiLL(-) (n = 52), patients with severe VP showed worse clinical outcomes, particularly for LBP and no differences in preoperative, postoperative, and improvement of LL were observed between two groups. CONCLUSION: Patients with DiLL (+) and DiLL (-) showed different clinical courses depending on VP severity at the non-fused discs after single-level TLIF.

Clonal Origin and Lineage Ambiguity in Mixed Neuroendocrine Carcinoma of the Uterine Cervix.

Small-cell neuroendocrine carcinoma (SCNEC) of the cervix is a rare disease characterized by a high incidence of mixed tumors with other types of cancer. The mechanism underlying this mixed phenotype is not well understood. This study established a panel of organoid lines from patients with SCNEC of the cervix and ultimately focused on one line, which retained a mixed tumor phenotype, both in vitro and in vivo. Histologically, both organoids and xenograft tumors showed distinct differentiation into either SCNEC or adenocarcinoma in some regions and ambiguous differentiation in others. Tracking single cells indicated the existence of cells with bipotential differentiation toward SCNEC and adenocarcinomas. Single-cell transcriptional analysis identified three distinct clusters: SCNEC-like, adenocarcinoma-like, and a cluster lacking specific differentiation markers. The expression of neuroendocrine markers was enriched in the SCNEC-like cluster but not exclusively. Human papillomavirus 18 E6 was enriched in the SCNEC-like cluster, which showed higher proliferation and lower levels of the p53 pathway. After treatment with anticancer drugs, the expression of adenocarcinoma markers increased, whereas that of SCNEC decreased. Using a reporter system for keratin 19 expression, changes in the differentiation of each cell were shown to be associated with the shift in differentiation induced by drug treatment. These data suggest that mixed SCNEC/cervical tumors have a clonal origin and are characterized by an ambiguous and flexible differentiation state.

Objective Assessment of Sleep Disorders in Patients with Lumbar Spinal Stenosis Using Wearable Trackers.

Introduction: Low-back pain causes sleep disorders, which impairs the quality of life (QOL) of patients. Sleep disorders are associated with lumbar spinal stenosis (LSS); however, the postoperative effects of LSS surgery on sleep disorders are unknown. This study aimed to assess sleep disorders in patients with LSS using wearable activity trackers and determine whether surgery improves sleep quality. Methods: A total of 39 patients scheduled for LSS surgery (mean age 71.1±8.7 years; 22 men and 17 women) were studied. Sleep disorders in the participants were objectively evaluated using a wearable Motionlogger Micro system. Sleep efficiency (SEf), mean active count (MAC), and wake after sleep onset (WASO) were measured before and 6 months following surgery. Furthermore, the patient-based outcomes of pain and QOL-related scores were measured and compared with those of healthy participants. The group with improved SEf following surgery was designated as "nonpoor sleepers," whereas the group that did not exhibit improvements was designated as "poor sleepers." The two groups were compared based on patient factors, patient-based questionnaires, and sleep disorder measurements. Results: The SEf and WASO were significantly worse in patients with LSS compared with healthy participants (P<0.05). Furthermore, the SEf in patients with LSS was associated with the Oswestry Disability Index scores. No improvement was observed in the SEf, MAC, and WASO before and after surgery. Evaluation of each case revealed 21 and 12 cases of nonpoor and poor sleepers, respectively. Preoperative low-back pain was significantly associated with improvement in postoperative sleep quality. Conclusions: Sleep disorders in patients with LSS were evaluated, and improvement in sleep disorders following surgery was associated with the intensity of preoperative low-back pain. Sleep disorders are associated with QOL disorders, suggesting that focusing on the treatment of sleep disorders is important in the management of patients with LSS.

Denosumab-associated Periprosthetic Atypical Femur Fracture: A Case Report.

Introduction: Denosumab is generally used for 5-10 years to treat postmenopausal osteoporosis. Although atypical fractures caused by bisphosphonate use are well known, reports of denosumab-associated femur fractures are rare. Case Report: Herein, a 75-year-old woman suffered an atypical periprosthetic femoral fracture 31 months after receiving denosumab. The fracture occurred transverse to the stem tip with lateral cortical thickening. The patient underwent revision surgery for conversion to a longer cemented stem. The fracture site healed 10 months after revision surgery. Conclusion: As far as we know, there have been no reports of a case on periprosthetic atypical femur fracture associated with denosumab. Our study shows the potential of periprosthetic atypical femoral fractures in patients using denosumab for a long time.

Ultra-Rapid and Specific Gelation of Collagen Molecules for Transparent and Tough Gels by Transition Metal Complexation.

Collagen is the most abundant protein in the human body and one of the main components of stromal tissues in tumors which have a high elastic modulus of over 50 kPa. Although collagen has been widely used as a cell culture scaffold for cancer cells, there have been limitations when attempting to fabricate a tough collagen gel with cells like a cancer stroma. Here, rapid gelation of a collagen solution within a few minutes by transition metal complexation is demonstrated. Type I collagen solution at neutral pH shows rapid gelation with a transparency of 81% and a high modulus of 1,781 kPa by mixing with K2 PtCl4 solution within 3 min. Other transition metal ions also show the same rapid gelation, but not basic metal ions. Interestingly, although type I to IV collagen molecules show rapid gelation, other extracellular matrices  do not exhibit this phenomenon. Live imaging of colon cancer organoids in 3D culture indicates a collective migration property with modulating high elastic modulus, suggesting activation for metastasis progress. This technology will be useful as a new class of 3D culture for cells and organoids due to its facility for deep-live observation and mechanical stiffness adjustment.

Risk Factors for Cage Subsidence in Minimally Invasive Lateral Corpectomy for Osteoporotic Vertebral Fractures.

Introduction: This study aims to investigate risk factors for cage subsidence following minimally invasive lateral corpectomy for osteoporotic vertebral fractures. Methods: Eight males and 13 females (77.2±6.0 years old) with osteoporotic vertebral fractures who underwent single corpectomy using a wide-footprint expandable cage with at least a 1-year follow-up were retrospectively included. The endplate cage (EC) angle was defined as the angle between the vertebral body's endplate and the cage's base on the cranial and caudal sides. A sagittal computed tomography scan was performed immediately after surgery and at the final follow-up, with cage subsidence defined as subsidence of ≥2 mm on the cranial or caudal side. Risk factors were analyzed by dividing cases into groups with (n=6) and without (n=15) cage subsidence. Results: No significant differences were noted in age, bone mineral density, number of fixed vertebrae, sagittal parameters, preoperative and final kyphosis angle, amount of kyphosis angle correction, bone union, screw loosening, and number of other vertebral fractures preoperatively and 1-year postoperatively between the two groups. No difference was noted in cranial EC angle, but a significant difference was noted in caudal EC angle in the group with (10.7±4.1°) and without (4.7±4.2°) subsidence (P=0.008). Logistic regression analysis with the dependent variable as presence or absence of subsidence showed that caudal EC angle (>7.5°) was a significant factor (odds ratio: 20, 95% confidence interval: 1.655-241.7, P=0.018). Conclusions: In minimally invasive lateral corpectomy for osteoporotic vertebral fractures, a cage tilted more than 7.5° to the caudal vertebral endplate is a risk factor for cage subsidence. The cage should be placed as perpendicular to the endplate as possible, especially to the caudal vertebral body, to avoid cage subsidence.

De-differentiation in cultures of organoids from luminal-type breast cancer is restored by inhibition of NOTCH signaling.

Estrogen receptor (ER) expression in breast cancer can change during progression and the treatment, but the mechanism has not been well studied. In this study, we successfully prepared organoids from samples obtained from 33 luminal-type breast cancer patients and studied their ER expression. The expression status was well maintained in primary organoids, whereas it decreased after passaging in most of the cases. In fact, the studied organoid lines were classified into those that retained a high level of ER expression (9%), those that completely lost it (9%), and those that repressed it to varying degrees (82%). In some cases, the ER expression was suddenly and drastically decreased after passaging. Marker protein immunohistochemistry revealed that after passaging, the differentiation status shifted from a luminal- to a basal-like status. Differentially expressed genes suggested the activation of NOTCH signaling in the passaged organoids, wherein a NOTCH inhibitor was able to substantially rescue the decreased ER expression and alter the differentiation status. Our findings suggest that the differentiation status of luminal-type cancer cells is quite flexible, and that by inhibiting the NOTCH signaling we can preserve the differentiation status of luminal-type breast cancer organoids.

Oxygen metabolism analysis of a single organoid for non-invasive discrimination of cancer subpopulations with different growth capabilities.

Heterogeneous nature is a pivotal aspect of cancer, rendering treatment problematic and frequently resulting in recurrence. Therefore, advanced techniques for identifying subpopulations of a tumour in an intact state are essential to develop novel screening platforms that can reveal differences in treatment response among subpopulations. Herein, we conducted a non-invasive analysis of oxygen metabolism on multiple subpopulations of patient-derived organoids, examining its potential utility for non-destructive identification of subpopulations. We utilised scanning electrochemical microscopy (SECM) for non-invasive analysis of oxygen metabolism. As models of tumours with heterogeneous subpopulations, we used patient-derived cancer organoids with a distinct growth potential established using the cancer tissue-originated spheroid methodology. Scanning electrochemical microscopy measurements enabled the analysis of the oxygen consumption rate (OCR) for individual organoids as small as 100 µm in diameter and could detect the heterogeneity amongst studied subpopulations, which was not observed in conventional colorectal cancer cell lines. Furthermore, our oxygen metabolism analysis of pre-isolated subpopulations with a slow growth potential revealed that oxygen consumption rate may reflect differences in the growth rate of organoids. Although the proposed technique currently lacks single-cell level sensitivity, the variability of oxygen metabolism across tumour subpopulations is expected to serve as an important indicator for the discrimination of tumour subpopulations and construction of novel drug screening platforms in the future.

Eribulin mesylate induces bone mass loss by promoting osteoclastic bone resorption in mice.

Over the past few decades, the clinical outcomes of patients with cancer have significantly improved mostly owing to the development of effective chemotherapeutic treatments. However, chronic health conditions such as bone mass loss and risk of fragility fractures caused by chemotherapy have also emerged as crucial issues in patients treated for cancer. In this study, we aimed to understand the effect of eribulin mesylate (ERI), a microtubule-targeting agent currently used to treat metastatic breast cancer and certain subtypes of advanced sarcomas, on bone metabolism in mice. The administration of ERI reduced bone mass in mice, mainly by promoting osteoclast activity. Gene expression analysis of skeletal tissues revealed no change in the expression levels of the transcripts for RANK ligand, one of the master regulators of osteoclastogenesis; however, the transcript levels of osteoprotegerin, which neutralizes RANK ligand, were significantly reduced in ERI-treated mice compared with those in vehicle-treated controls, indicating a relative increase in RANK ligand availability after ERI treatment. In line with the increased bone resorption in ERI-treated mice, we found that zoledronate administration effectively suppressed bone loss in these mice. These results reveal a previously unrecognized effect of ERI on bone metabolism and suggest the application of bisphosphonates for patients with cancer undergoing treatment with ERI.

Inhibition of the bone morphogenetic protein pathway suppresses tumor growth through downregulation of epidermal growth factor receptor in MEK/ERK-dependent colorectal cancer.

The bone morphogenetic protein (BMP) pathway promotes differentiation and induces apoptosis in normal colorectal epithelial cells. However, its role in colorectal cancer (CRC) is controversial, where it can act as context-dependent tumor promoter or tumor suppressor. Here we have found that CRC cells reside in a BMP-rich environment based on curation of two publicly available RNA-sequencing databases. Suppression of BMP using a specific BMP inhibitor, LDN193189, suppresses the growth of select CRC organoids. Colorectal cancer organoids treated with LDN193189 showed a decrease in epidermal growth factor receptor, which was mediated by protein degradation induced by leucine-rich repeats and immunoglobulin-like domains protein 1 (LRIG1) expression. Among 18 molecularly characterized CRC organoids, suppression of growth by BMP inhibition correlated with induction of LRIG1 gene expression. Notably, knockdown of LRIG1 in organoids diminished the growth-suppressive effect of LDN193189. Furthermore, in CRC organoids, which are susceptible to growth suppression by LDN193189, simultaneous treatment with LDN193189 and trametinib, an FDA-approved MEK inhibitor, resulted in cooperative growth inhibition both in vitro and in vivo. Taken together, the simultaneous inhibition of BMP and MEK could be a novel treatment option in CRC cases, and evaluating in vitro growth suppression and LRIG1 induction by BMP inhibition using patient-derived organoids could offer functional biomarkers for predicting potential responders to this regimen.

Tracking the growth fate of single cells and isolating slow-growing cells in human colorectal cancer organoids.

Patient-derived tumor organoids are three-dimensionally cultured cancer cells that enable a suitable platform for studying heterogeneity and plasticity of cancer. We present a protocol for tracking the growth fate of single cells and isolating slow-growing cells in human colorectal cancer organoids. We describe steps for organoid preparation and culturing using the cancer-tissue-originating spheroid method, maintaining cell-cell contact throughout. We then detail a single-cell-derived spheroid-forming and growth assay, confirming single-cell plating, monitoring growth over time, and isolating slow-growing cells. For complete details on the use and execution of this protocol, please refer to Coppo et al.1.

Effect of Preoperative Severity and Location of Lumbar Intervertebral Disc Vacuum Phenomenon on Surgical Outcomes After Single-Level Transforaminal Lumbar Interbody Fusion.

OBJECTIVE: This study aimed to examine whether preoperative severity and location of lumbar intervertebral disc vacuum phenomenon (VP) influence surgical outcomes after single-level transforaminal lumbar interbody fusion. METHODS: We included 106 patients (age, 67.4 ± 10.4 years; 51 male/55 female) with lumbar degenerative diseases, who were treated with single-level transforaminal lumbar interbody fusion. Severity of VP (SVP) score was measured preoperatively. SVP score at fused disc was used as SVP (FS) score and at nonfused discs was used as SVP (non-FS) score. Surgical outcomes were assessed using the Oswestry Disability Index (ODI) and visual analog scale (VAS; low back pain (LBP), lower extremity pain, numbness, LBP in motion, in standing, and in sitting). The patients were divided into severe VP (FS or non-FS) and mild VP (FS or non-FS) groups, and surgical outcomes were compared between the 2 groups. Correlations between each SVP score and surgical outcomes were analyzed. RESULTS: There were no differences in surgical outcomes between the severe VP (FS) and mild VP (FS) groups. Postoperative ODI, VAS score for LBP, lower extremity pain, numbness, and LBP in standing were significantly worse in the severe VP (non-FS) group than in the mild VP (non-FS) group. SVP (non-FS) scores significantly correlated with postoperative ODI, VAS score for LBP, lower extremity pain, numbness, and LBP in standing; however, SVP (FS) scores did not correlate with any surgical outcomes. CONCLUSIONS: Preoperative SVP at fused disc is not associated with surgical outcomes; however, SVP at nonfused discs is correlated with clinical outcomes.