RESUMO
Mesothelial cells are arranged as a monolayer on covering membranes that invest surfaces of body cavities like the pleura and peritoneum. Primary human mesothelial cell (HMC) cultures are needed for studying mesothelial cell homeostasis and developing disease models, such as wound healing or cancers. Remarkably, there is a paucity of useable HMC lines that are currently available that faithfully recapitulate normal in vivo phenotypic characteristics. Here, we present a strategy to recover HMC from human pleural tissue and to immortalize them for extended in vitro culturing. Human pleural membrane was harvested by minimally invasive surgical techniques. HMC were isolated using a two-step process combining explant cellular outgrowth from biopsy tissue and flow cytometry based on cell surface expression of cadherin-1 and CD71. Cell cultures were generated after lentiviral transfection with human telomerase. The new HMC cultures retain the same phenotypic traits and physiologic features as their in vivo counterparts, yet they can be adapted for short-term or long-term culture in large-scale in vitro experimentation. In particular, we generated a new HMC line harboring a germline mutation in breast cancer type-1-associated protein-1 (BAP1), a causal tumor suppressor gene, that could be instrumental to malignant mesothelioma research. Patient-specific, normal HMC may serve as novel discovery tools allowing more powerful research models of both normal physiology and disease processes. Our surgically driven approach leads to a limitless resource of novel mesothelial cell cultures.
Assuntos
Epitélio/patologia , Neoplasias Pulmonares/patologia , Mesotelioma/patologia , Pleura/patologia , Neoplasias Pleurais/patologia , Biópsia/métodos , Humanos , Mesotelioma Maligno , Neoplasias Pleurais/metabolismo , Células Tumorais Cultivadas , Proteínas Supressoras de Tumor/metabolismo , Ubiquitina Tiolesterase/metabolismoRESUMO
OBJECTIVE: Determine if superior canal dehiscence (SCD) found on flat-panel CT increases the risk for other defects in the otic capsule. STUDY DESIGN: Retrospective cohort study. SETTING: Tertiary care center. PATIENTS: One hundred ears (50 with SCD and 50 matched controls without SCD). INTERVENTIONS: Flat-panel CT imaging. MAIN OUTCOME MEASURES: (1) Prevalence of other dehiscences in SCD ears, (2) dehiscences in controls, and (3) otic capsule thickness in other reported dehiscence locations (cochlea-carotid, lateral semicircular canal [SCC] and mastoid, facial nerve-lateral SCC, vestibular aqueduct, posterior SCC-jugular bulb, posterior SCC-posterior fossa). Between-group comparisons were considered significant at p < 0.007 after applying the Bonferroni correction for multiple comparisons. RESULTS: Not including the SCD, there was a mean of 0.04 additional dehiscences in the SCD group (n = 2/50, 4%) and 0.04 non-SCD dehiscences in the controls (n = 2/50, 4%, p > 0.007). In the SCD group, there was one dehiscence between the cochlea and carotid artery and one between the posterior SCC and posterior fossa. The control group had one enlarged vestibular aqueduct and one dehiscence between the facial nerve and lateral SCC. As a group, SCD ears had wider vestibular aqueducts (0.68 ± 0.20 vs 0.51 ± 0.30 mm, p < 0.007) and thinner bone between the posterior SCC and posterior fossa (3.12 ± 1.43 vs 4.34 ± 1.67 mm, p < 0.007). The bone between the facial nerve and lateral SCC was thicker in SCD ears (0.77 ± 0.23 vs 0.55 ± 0.27 mm, p < 0.007) and no different for cochlea-carotid, and lateral SCC and mastoid (p > 0.007). CONCLUSIONS: SCD does not increase the likelihood of a second dehiscence in the same otic capsule. SCD patients may have congenitally thinner otic capsule bones compared to controls, particularly near the posterior SCC, where the vestibular aqueduct may be enlarged.
Assuntos
Deiscência do Canal Semicircular , Canais Semicirculares , Tomografia Computadorizada por Raios X , Humanos , Estudos Retrospectivos , Masculino , Feminino , Pessoa de Meia-Idade , Canais Semicirculares/diagnóstico por imagem , Canais Semicirculares/patologia , Adulto , Deiscência do Canal Semicircular/diagnóstico por imagem , Deiscência do Canal Semicircular/patologia , Idoso , Estudos de Coortes , Aqueduto Vestibular/diagnóstico por imagem , Aqueduto Vestibular/patologia , Aqueduto Vestibular/anormalidades , Cóclea/diagnóstico por imagem , Cóclea/patologia , Processo Mastoide/diagnóstico por imagem , Processo Mastoide/patologiaRESUMO
OBJECTIVE: Use microscopic video-based tracking of laryngeal surgical instruments to investigate the effect of robot assistance on instrument tremor. STUDY DESIGN: Experimental trial. SETTING: Tertiary Academic Medical Center. METHODS: In this randomized cross-over trial, 36 videos were recorded from 6 surgeons performing left and right cordectomies on cadaveric pig larynges. These recordings captured 3 distinct conditions: without robotic assistance, with robot-assisted scissors, and with robot-assisted graspers. To assess tool tremor, we employed computer vision-based algorithms for tracking surgical tools. Absolute tremor bandpower and normalized path length were utilized as quantitative measures. Wilcoxon rank sum exact tests were employed for statistical analyses and comparisons between trials. Additionally, surveys were administered to assess the perceived ease of use of the robotic system. RESULTS: Absolute tremor bandpower showed a significant decrease when using robot-assisted instruments compared to freehand instruments (P = .012). Normalized path length significantly decreased with robot-assisted compared to freehand trials (P = .001). For the scissors, robot-assisted trials resulted in a significant decrease in absolute tremor bandpower (P = .002) and normalized path length (P < .001). For the graspers, there was no significant difference in absolute tremor bandpower (P = .4), but there was a significantly lower normalized path length in the robot-assisted trials (P = .03). CONCLUSION: This study demonstrated that computer-vision-based approaches can be used to assess tool motion in simulated microlaryngeal procedures. The results suggest that robot assistance is capable of reducing instrument tremor.