A Modified Magnified Analysis of Proteome (MAP) Method for Super-Resolution Cell Imaging that Retains Fluorescence

Biological systems consist of a variety of distinct cell types that form functional networks. Super-resolution imaging of individual cells is required for better understanding of these complex systems. Direct visualization of 3D subcellular and nano-scale structures in cells is helpful for the interpretation of biological interactions and system-level responses. Here we introduce a modified magnified analysis of proteome (MAP) method for cell super-resolution imaging (Cell-MAP) which preserves cell fluorescence. Cell-MAP expands cells more than four-fold while preserving their overall architecture and three-dimensional proteome organization after hydrogel embedding. In addition, Optimized-Cell-MAP completely preserves fluorescence and successfully allows for the observation of tagged small molecular probes containing peptides and microRNAs. Optimized-Cell-MAP further successfully applies to the study of structural characteristics and the identification of small molecules and organelles in mammalian cells. These results may give rise to many other applications related to the structural and molecular analysis of smaller assembled biological systems.ope

Three-Dimensional Printed Mesh-Cage Replacement for a Metastatic Spinal Tumor and a Vertebral Compression Fracture: A Report of Two Cases

Harms mesh cages are commonly used in the treatment of metastatic spinal tumors by spondylectomy with cage insertion and posterior screw fixation. These cages are even useful for the surgical reconstruction of spinal deformities with mesh-cage replacement after vertebral column resection (VCR). Mesh cages can be customized by 3-dimensional (3D) printing based on the angle between the endplates for the requisite weight-bearing capacity and fit. This study reports two cases wherein a 75-year-old man and a 63-year-old woman were treated with reconstructive spondylectomy and mesh-cage replacement, followed by posterior screw fixation. The 75-year-old man was initially diagnosed with an L4 metastatic non-small cell lung carcinoma accompanied by a pathologic fracture and severe paralysis. The 63-year-old woman had slipped 13 years before and sustained a T11-L1 compression fracture that was aggravated 6 months before this admission, with back pain and numbness in both soles. Herein, we describe the treatment approach with instrumentation using a customized 3D-printed mesh cage after total spondylectomy for the metastatic spinal tumor and after VCR for deformity repair surgery in the respective cases. Our application of a 3D-printed mesh cage in surgery for a metastatic tumor is the first report of such treatment in Korea. Although the use of customized 3D-printed cages for a metastatic spinal tumor and a severe compression fracture was effective, challenges remain regarding fitting, manufacturing time, and costs.ope

The Role and Future of Endoscopic Spine Surgery: A Narrative Review

Many types of surgeries are changing from conventional to minimally invasive techniques. Techniques in spine surgery have also changed, with endoscopic spine surgery (ESS) becoming a major surgical technique. Although ESS has advantages such as less soft tissue dissection and normal structure damage, reduced blood loss, less epidural scarring, reduced hospital stay, and earlier functional recovery, it is not possible to replace all spine surgery techniques with ESS. ESS was first used for discectomy in the lumbar spine, but the range of ESS has expanded to cover the entire spine, including the cervical and thoracic spine. With improvements in ESS instruments (optics, endoscope, endoscopic drill and shaver, irrigation pump, and multiportal endoscopic), limitations of ESS have gradually decreased, and it is possible to apply ESS to more spine pathologies. ESS currently incorporates new technologies, such as navigation, augmented and virtual reality, robotics, and 3-dimentional and ultraresolution visualization, to innovate and improve outcomes. In this article, we review the history and current status of ESS, and discuss future goals and possibilities for ESS through comparisons with conventional surgical techniques.ope

What Affects Segmental Lordosis of the Surgical Site after Minimally Invasive Transforaminal Lumbar Interbody Fusion?

Purpose: This study was undertaken to identify factors that affect segmental lordosis (SL) after minimally invasive transforaminal lumbar interbody fusion (MIS-TLIF) by comparing patients whose postoperative SL increased with those whose decreased. Materials and methods: Fifty-five patients underwent MIS-TLIF at our institute from January 2018 to September 2019. Demographic, pre- and postoperative radiologic, and cage-related factors were included. Statistical analyses were designed to compare patients whose SL increased with decreased after surgery. Results: After surgery, SL increased in 34 patients (group I) and decreased in 21 patients (group D). The index level, disc lordosis, SL, lumbar lordosis, proximal lordosis (PL), and Y-axis position of the cage (Yc) differed significantly between groups I and D. The cage in group I was more anterior than that in group D (Yc: 55.84% vs. 51.24%). Multivariate analysis showed that SL decreased more significantly after MIS-TLIF when the index level was L3/4 rather than L4/5 [odds ratio (OR): 0.46, p=0.019], as preoperative SL (OR: 0.82, p=0.037) or PL (OR: 0.68, p=0.028) increased, and as the cage became more posterior (OR: 1.10, p=0.032). Conclusion: Changes in SL after MIS-TLIF appear to be associated with preoperative SL and PL, index level, and Yc. An index level at L4/5 instead of L3/4, smaller preoperative SL or PL, and an anterior position of the cage are likely to result in increased SL after MIS-TLIF. Materials and methods: Fifty-five patients underwent MIS-TLIF at our institute from January 2018 to September 2019. Demographic, pre- and postoperative radiologic, and cage-related factors were included. Statistical analyses were designed to compare patients whose SL increased with decreased after surgery. Results: After surgery, SL increased in 34 patients (group I) and decreased in 21 patients (group D). The index level, disc lordosis, SL, lumbar lordosis, proximal lordosis (PL), and Y-axis position of the cage (Yc) differed significantly between groups I and D. The cage in group I was more anterior than that in group D (Yc: 55.84% vs. 51.24%). Multivariate analysis showed that SL decreased more significantly after MIS-TLIF when the index level was L3/4 rather than L4/5 [odds ratio (OR): 0.46, p=0.019], as preoperative SL (OR: 0.82, p=0.037) or PL (OR: 0.68, p=0.028) increased, and as the cage became more posterior (OR: 1.10, p=0.032). Conclusion: Changes in SL after MIS-TLIF appear to be associated with preoperative SL and PL, index level, and Yc. An index level at L4/5 instead of L3/4, smaller preoperative SL or PL, and an anterior position of the cage are likely to result in increased SL after MIS-TLIF.ope

The Technical Feasibility of Unilateral Biportal Endoscopic Decompression for The Unpredicted Complication Following Minimally Invasive Transforaminal Lumbar Interbody Fusion: Case Report

Minimally invasive techniques for transforaminal lumbar interbody fusion (MIS-TLIF) are advantageous because they allow for sufficient surgical exposure and fewer complications through a smaller incision than conventional TLIF. It could be difficult to maintain minimally invasive spine surgery following the unexpected complications after MIS-TLIF. Because MIS-TLIF is usually done via a paramedian small incision with posterior fusion using screws and rods, visualization of the surgical field is limited, and it is difficult to directly assess the neural structure without removing instrumentation. Unilateral biportal endoscopic decompression (UBE) is a rapidly growing surgical method using two 1-cm incisions that are 2 to 3 cm apart. We would like to suggest UBE as an option for immediate reoperation after MIS-TLIF because it has the advantages of targeting pathologic regions and a wide field of visualization through small wounds. The operation is independent of the existing incision from MIS-TLIF, enabling immediate revision surgery without the removal of the screws and rods. UBE has the advantages of targeting specific surgical regions and providing a wide visualization of the operation field through small incisions. UBE can be very useful for discectomy or decompression surgery as well as in immediate reoperation after MIS-TLIF.ope

Multiplexed and scalable super-resolution imaging of three-dimensional protein localization in size-adjustable tissues

The biology of multicellular organisms is coordinated across multiple size scales, from the subnanoscale of molecules to the macroscale, tissue-wide interconnectivity of cell populations. Here we introduce a method for super-resolution imaging of the multiscale organization of intact tissues. The method, called magnified analysis of the proteome (MAP), linearly expands entire organs fourfold while preserving their overall architecture and three-dimensional proteome organization. MAP is based on the observation that preventing crosslinking within and between endogenous proteins during hydrogel-tissue hybridization allows for natural expansion upon protein denaturation and dissociation. The expanded tissue preserves its protein content, its fine subcellular details, and its organ-scale intercellular connectivity. We use off-the-shelf antibodies for multiple rounds of immunolabeling and imaging of a tissue's magnified proteome, and our experiments demonstrate a success rate of 82% (100/122 antibodies tested). We show that specimen size can be reversibly modulated to image both inter-regional connections and fine synaptic architectures in the mouse brain.ope

Comparative Assessment of Different Percutaneous Endoscopic Interlaminar Lumbar Discectomy (PEID) Techniques

BACKGROUND: Percutaneous endoscopic lumbar discectomy is a common surgical treatment for lumbar disc herniation, and percutaneous endoscopic interlaminar lumbar discectomy (PEID) is commonly used for direct decompression of L5-S1. Like microdiscectomy, recurrence of herniation after endoscopic discectomy is an important problem. In this study, we aimed to decrease the recurrence after PEID using a new surgical technique. OBJECTIVES: We propose a new surgical technique for reducing the recurrence after PEID for lumbar disc herniation. The new technique uses annular sealing after fragmentectomy. We compared clinical results and recurrent lumbar disc herniation (had radiculopathy and confirmed by MRI) between patients who underwent surgery with and without annular sealing during PEID. STUDY DESIGN: Retrospective cohort study of patients undergoing PEID. METHODS: A total of 224 patients with radiculopathy due to L5-S1 disc herniation who were treated by PEID with (91 patients) or without annular sealing (133) were included in this study. We compared the demographic characteristics (age, sex, height, weight, BMI, smoking status, and occupation), clinical results, and recurrence rates between the 2 groups. We classified recurrence according to time period (early recurrence ≤ 6 months, late recurrence > 6 months). RESULTS: The study groups were demographically similar, and substantial improvement in clinical results was noted. There were 5 recurrences (5.5%) (2 early, 3 late recurrences) in the group with annular sealing, and 18 (13.5%) (13 early, 5 late recurrences) in the group without annular sealing. Early recurrence rates were significantly higher in the group without sealing (2 vs. 13, P = 0.029). Increasing age was associated with overall recurrence (P = 0.004) and late recurrence (P = 0.008), while operative technique correlated with early recurrence (P = 0.026). LIMITATIONS: First, this study incorporates a retrospective design. Second, the operations were performed by 2 surgeons. Additionally, this is relatively a short-term follow-up study (mean 19.5 ± 5.0 months). CONCLUSIONS: Though a learning curve is needed in order to become familiar with PEID, recurrence after PEID was associated with advanced age, and PEID with annular sealing resulted in lower early recurrence rates than without annular sealing. Thus, PEID with annular sealing may be a useful technique for reducing early recurrence.ope

3D Printer Application for Endoscope-Assisted Spine Surgery Instrument Development: From Prototype Instruments to Patient-Specific 3D Models

Developing new surgical instruments is challenging. While making surgical instruments could be a good field of application for 3D printers, attempts to do so have proven limited. We designed a new endoscope-assisted spine surgery system, and using a 3D printer, attempted to create a complex surgical instrument and to evaluate the feasibility thereof. Developing the new surgical instruments using a 3D printer consisted of two parts: one part was the creation of a prototype instrument, and the other was the production of a patient model. We designed a new endoscope-assisted spine surgery system with a cannula for the endoscope and working instruments and extra cannula that could be easily added. Using custom-made patient-specific 3D models, we conducted discectomies for paramedian and foraminal discs with both the newly designed spine surgery system and conventional tubular surgery. The new spine surgery system had an extra portal that can be well bonded in by a magnetic connector and greatly expanded the range of access for instruments without unnecessary bone destruction. In foraminal discectomy, the newly designed spine surgery system showed less facet resection, compared to conventional surgery. We were able to develop and demonstrate the usefulness of a new endoscope-assisted spine surgery system relying on 3D printing technology. Using the extra portal, the usability of endoscope-assisted surgery could be greatly increased. We suggest that 3D printing technology can be very useful for the realization and evaluation of complex surgical instrument systems.ope

Comparison Between 3-Dimensional-Printed Titanium and Polyetheretherketone Cages: 1-Year Outcome After Minimally Invasive Transforaminal Interbody Fusion

Objective: Three-dimensional (3D)-printed titanium implants have been developed recently, but the utility is not yet proven. The aim of this study was to compare 3D-printed titanium and polyetheretherketone (PEEK) implants after minimally invasive transforaminal lumbar interbody fusion (MIS-TLIF). Methods: Between October 2018 and September 2021, we retrospectively analyzed 83 patients who underwent single-level MIS-TLIF (3D-printed titanium, 40; PEEK, 43). Radiologic parameters were assessed with x-ray and computed tomography (CT) at postoperative 1 week, 6 months, and 1 year. Clinical status was evaluated using Oswestry Disability Index, visual analogue scale score, and Bridwell fusion grading was assessed on 6-month and 1-year postoperative CT. Results: There were no differences between the 2 groups in demographics and clinical outcomes. At 1-year of follow-up, the reported 3D-printed titanium fusion grades were grade I: 77.5% (31 patients), grade II: 17.5% (7 patients), and grade III: 5% (2 patients). The PEEK fusion grades were grade I: 51.2% (22 patients), grade II: 41.9% (18 patients), and grade III: 7.0% (3 patients). For overall fusion rate (grade I + II), there was no difference between the 2 cages (95.0% vs. 93.0%, p = 0.705), but grade I was reported at a higher incidence in 3D-printed titanium than PEEK (77.5% vs. 51.2%, p = 0.013). There was no difference between cages based on subsidence and complications. Conclusion: There were no significant differences in the overall fusion rate for MIS-TLIF surgery between 3D-printed titanium and PEEK, but the fusion grade was better in 3D-printed titanium than in PEEK. Long-term follow-up is required to verify the effectiveness.ope

Surgical Outcomes of Dysphagia Provoked by Diffuse Idiopathic Skeletal Hyperostosis in the Cervical Spine

Purpose: This study aimed to predict the surgical outcomes of diffuse idiopathic skeletal hyperostosis (DISH)-related dysphagia (DISH-phagia) and to evaluate the importance of prevertebral soft tissue thickness (PVST). Materials and methods: In total, 21 surgeries (anterior osteophytectomy or anterior cervical decompression and fixation) were included in this study for DISH-phagia from 2003 to 2019. Clinical outcomes were assessed using the Dysphagia Outcome and Severity Scale (DOSS) preoperatively, at 1 month postoperatively, and last follow up (mean 29.5 months). PVST was measured using lateral plain radiographs. Paired t-test and Spearman's correlation test was used to identify relationships between various PVST indices and DOSS. Results: Comparisons were made from 17 patients out of 21, in which the record had all of three measurements. The narrowest PVST preoperatively was 2.55±0.90 mm, with a DOSS score of 4.47±1.61, and that at 1 month after surgery was 5.02±2.33 mm, with a DOSS score of 6.12±1.32. At last follow up, PVST and DOSS values were 3.78±0.92 mm and 5.82±1.34, and three patients experienced symptom relapse. Significant relationships were found between PVST and DOSS at all time points: before surgery (R=0.702, p<0.001), 1 month after surgery (R=0.539, p=0.012), and last follow up (R=0.566, p=0.020). Conclusion: Surgical removal of anterior osteophytes is an effective treatment option for DISH-phagia, and PVST is a useful parameter in DISH-phagia. The goal of DISH surgery should be to remove DISH as much as possible to ensure sufficient PVST postoperatively.ope