Cadaveric Tissue Research Articles

Neuroendoscopy Training

AbstractNeuroendoscopy can be learnt by assisting or doing live human surgery, cadaver dissection with or without augmented pulsatile vessel and cerebrospinal fluid (CSF) perfusion, and practicing on live animal, dead animal model, synthetic models, three-dimensional printing model with or without augmentation with animal, cadaver tissue, pulsatile vessel and reconstructed CSF model, virtual reality (VR) simulator, and hybrid simulators (combined physical model and VR model). Neurosurgery skill laboratory with basic and advanced learning should be there in all teaching hospitals. Skills can be transferred from simulation model or VR to cadaver to live surgery. Staged learning (first with simple model to learn basic endoscopic technique, then animal model, and then augmented cadavers) is the preferred method of learning. Although most surveys favor live surgery and practice on animal models and cadavers as the most preferred training model now, in future VR may also become a favored method of learning. This article is based on our experience in over 10,000 neuroendoscopic surgeries, and feedback from over 950 neuroendoscopic fellows or consultants who attended workshops conducted every 6 monthly since 2010. A literature search was done on PubMed and Google Scholar using (neuroendoscopy) AND (learning), and (neuroendoscopy) AND (training), which resulted in 121 and 213 results, respectively. Out of them, 77 articles were finally selected for this article. Most of the training programs typically focus on microneurosurgical training. There is lack of learning facilities for neuroendoscopy in most centers. Learning of neuroendoscopy differs greatly from microneurosurgery; switching from microneurosurgery to neuroendoscopy can be challenging. Postgraduate training centers should have well-equipped neuroendoscopy skill laboratory and the surgical educational curriculum should include neuroendoscopy training. Learning endoscopy is about taking advantages of the technique and overcoming the limitations of endoscopy by continuous training.

Rhinoplasty Training: An Innovative Learning Approach Using Chicken Breast.

Mastering rhinoplasty surgical techniques requires extensive learning involving theoretical studies, internships, cadaver laboratories, simulators, nonhuman models, and hands-on practice. A novel method using chicken breast simplifies skill development, offering flexibility and accessibility. Training on chicken breast allows surgeons to refine skills through various techniques and grafts. This study focuses on preservation techniques using chicken as a model. Surgical sequences simulate nasal structures using prepared chicken breasts. Documentation through media illustrates procedures from basic to advanced, using materials akin to traditional rhinoplasty but in reduced quantities. Chicken breast tissue, especially sternal cartilage, offers a suitable substrate for practicing techniques and grafting without live human or cadaveric tissue. This method provides an efficient, safe, and cost-effective approach to learning rhinoplasty.

Mechanical evaluation of human cadaveric lumbar soft tissues suggests possible physiological stress shielding within musculoskeletal soft tissues by the thoracolumbar fascia.

This study investigates the potential for stress shielding within musculoskeletal soft tissues through analysis of stress distributions between lumbar fascial and muscle tissues via mechanical testing. Using a custom apparatus, 51 posterior thoracolumbar fascia (TLF) samples and 18 erector spinae (ES) cadaveric samples underwent tensile testing involving three loading-unloading cycles, followed by loading to 6% strain, to mechanically characterize samples. Parallel tensile testing using 20 pairs of two TLF samples, and seven pairs of TLF and ES samples was then conducted for stress distribution analysis between tissues. P<0.05 was deemed significant. The TLF and ES exhibited an average elastic modulus of 150.9MPa and 0.6MPa, respectively. At 6% strain, parallel testing of the TLF pairs yielded an average tensile stress of 8.4MPa and 1.7MPa (p<0.001) exhibited by the stiffer and less stiff TLF samples, respectively. Similarly, TLF-ES parallel testing resulted in average tensile stresses of 7.1MPa and 0.07MPa of the TLF and ES (p<0.002). Results suggest elevated loading towards stiffer TLF samples relative to less stiff TLF and ES samples. In soft tissues affected by LBP, skewed stress distributions may result in the TLF withstanding the majority of stress, yielding cyclical stress shielding that may contribute to and/or promote LBP. This novel study demonstrates a potential load allocation bias towards the TLF, laying the foundation for stress shielding within lumbar musculoskeletal soft tissues affected by degenerative musculoskeletal conditions.

Teaching medical students ultrasound-guided needle aspiration of synthetic cysts: effect of a formalin-embalmed cadaver simulation model

BackgroundStandard training for ultrasound-guided cyst needle aspiration is currently performed on live patients during residency. This practice presents risk of iatrogenic injury to patients and provides a high-stress learning environment for medical trainees. Simulation training using synthetic cysts in a formalin-embalmed cadaver model may allow for realistic, practical, and effective training free from patient risk.MethodsThirty first-year medical students viewed an orientation video, then attended a skills workshop to perform cyst needle aspiration in formalin-embalmed cadaver tissue under ultrasound guidance. Participants were randomly assigned to one of three ultrasound-trained instructor-types which included a medical student, clinical anatomist, or an ultrasound fellowship trained emergency medicine physician. After training, participants underwent a 5-min skills test to assess their ability to drain a synthetic cyst independently. Pre- and post-training self-confidence surveys were administered.ResultsUltrasound images of synthetic cysts in formalin-embalmed tissue were clear and realistic in appearance, and sonographic needle visualization was excellent. Participants took an average of 161.5 s and 1.9 attempts to complete the procedure. Two of the 30 participants could not complete the procedure within the time limit. Participants’ self-reported confidence with respect to all aspects of the procedure significantly increased post-training. Mean confidence scores rose from 1.2 (95% CI 0.96 to 1.39) to 4.4 (95% CI 4.09 to 4.53) (P < 0.0001) Procedure time, number of attempts, performance scores, and self-confidence outcomes were not significantly affected by instructor type.ConclusionsThe use of synthetic cysts in formalin-embalmed cadaveric tissue is feasible, realistic, and efficacious for the teaching of ultrasound-guided needle aspiration to novice medical trainees. This simulation training method can be delivered effectively by multiple instructor types and may allow medical trainees to increase their tactical skill and self-confidence prior to performing ultrasound-guided cyst needle aspiration on live patients.

Insects on Decomposing Pigs and their Forensic Importance at a University in Nigeria

The use of insects to estimate post-mortem interval (PMI) of cadavers by forensic entomologists, is an innovative science enshrined in the medico-criminal system of the developed world. The science is dependent on the biology of insect species that are found in decomposing cadavers. Thus, insect species that are found on decomposing pigs as model to human cadaver, was studied during the rainy and dry seasons of 2021 in a University forest in Nigeria. The decomposition stages (fresh through skeletonization) of the pig cadavers were monitored, and insects found on them were collected with a sweep net, visually observed, and handpicked. They were identified morphologically using insect identification keys with the aid of a dissecting microscope. The tissues of the pig cadavers decomposed completely in 14 and 10 days, respectively, for dry and rainy seasons. Twenty-seven insect species belonging to the orders; Diptera, Coleoptera, and Hymenoptera were identified. The dipterans consist of the families Calliphoridae, Muscidae, Ulidiidae, and Stratiomyidae. The coleopteran families are the Carabidae, Chrysomelidae, Cleridae, Dermestidae, Histeridae, Scarabaeidae, and Staphylinidae, while Formicidae is the only family for the hymenopteran. The dipteran species were the first group to visit the cadavers during fresh, bloated, and wet decay stages while the coleopterans visited during wet and dry decay stages thus, creating a succession pattern on the cadavers. The insect species that aided the cadaver decomposition at different stages are discussed as forensic indicators, which could be useful as PMI estimators of the study area.

Sustainable varicose vein therapy using functionalized hydrogels derived solely from livestock waste

Developing sustainable and effective treatments for chronic venous insufficiency (CVI) is crucial. In this study, we propose an innovative restorative approach utilizing hydrogels derived from the decellularized extracellular matrix (dECM) of cadaveric vascular tissues, adipose-derived stem cells (ADSCs), and gold nanoparticles (AuNPs). This therapeutic method leverages waste valorization by repurposing discarded cadaveric tissues from slaughterhouse livestock. The dECM hydrogels, enriched with ADSCs and AuNPs, offer a biocompatible scaffold that supports cellular differentiation and vascular integrity. Our approach addresses the limitations of current allo-, auto-, and xenograft methods by enhancing integration and functionality while potentially reducing costs through sustainable practices. This study explores functionalized hydrogel formulation solely generated from agri-food waste, gelation mechanisms, and preliminary cost-effectiveness, presenting a promising new avenue for treating early-stage varicose veins that can ultimately be translated to human models using discarded tissues.

Mass Spectrometry Imaging of the Subchondral Bone in Osteoarthritis Reveals Tissue Remodeling of Extracellular Matrix Proteins that Precede Cartilage Loss.

Osteoarthritis (OA) of the knee is a degenerative condition of the skeletal extracellular matrix (ECM) marked by the loss of articular cartilage and subchondral bone homeostasis. Treatments for OA in the knee beyond full joint replacement are lacking primarily due to gaps in molecular knowledge of the biological drivers of disease. Here, Mass Spectrometry Imaging (MSI) enabled molecular spatial mapping of the proteomic landscape of human knee tissues. Histologic sections of human tibial plateaus from OA patients and cadaveric controls were treated with collagenase III to target ECM proteins prior to imaging using a timsTOF fleX mass spectrometer (Bruker) for matrix-assisted laser desorption ionization (MALDI)-MSI of bone and cartilage proteins in human knees. Spatial MSI data of the knee, using sections of the tibial plateau from non-arthritic, cadaveric donors or from knee replacement patients with medial OA were processed and automatically segmented identifying distinct areas of joint damage. ECM peptide markers compared either OA to cadaveric tissues or OA medial to OA lateral. Not only did candidate peptides distinguish OA relative to intact cartilage, but also emphasized a significant spatial difference between OA and intact subchondral bone (AUROC >0.85). Overall, 31 peptide candidates from ECM proteins, including COL1A1, COL3A1, and unanticipated detection of collagens COL6A1 and COL6A3 in adult bone, exhibited significantly elevated abundance in diseased tissue. Highly specific hydroxyproline-containing collagens dominated OA subchondral bone directly under regions of lost cartilage revealing dramatic tissue remodeling providing molecular details on the progression of joint degeneration in OA. The identification of specific spatial markers for the progression of subchondral bone degeneration in OA advances our molecular understanding of coupled deterioration of joint tissues.

Global trends and insights in ethical statements regarding the utilization of human cadaveric tissues for biomechanical research from 2017 to 2022: a bibliometric analysis.

Human cadaveric tissues are essential for biomechanical research, however, omitting statements regarding the ethical implications remains very common in many recent articles of biomechanics. We systematically searched PubMed for research articles published from 2017 to 2022 to identify all relevant studies (n=1850). Information on the characteristics of the cadaveric specimens and the acquisition of ethical approval was collected. The included articles were categorized as grade A (n=757), grade B (n=308), and grade C (n=785) according to our predetermined criteria. Additionally, we analyzed whether the reporting status of ethical approval varied based on publishing year, journals, regions, and research settings. No significant improvement was observed over the study period. Disparities in ethical reporting standards were identified among different regions and journals. This is the first bibliometric analysis that offers an overview of publications in biomechanical research that use human cadaveric samples. It serves as a reference for interested researchers and journal readers to discuss the associated ethical issues.

Energy-based devices and hyaluronic acid filler, polymer filler, and threads: Cadaveric study.

The objective of this experiment was to investigate the thermal effects on hyaluronic acid fillers, PCL fillers, and PDO threads when exposed to controlled heat. This study aims to provide insights into how these materials respond to thermal energy, which is crucial for safe and effective cosmetic procedures involving combined modalities. Cadaveric tissue was utilized to simulate clinical conditions. Hyaluronic acid fillers were injected at approximately 1 mm and 5 mm thicknesses, with variations in G' value (high and low). PCL fillers were similarly injected in 1 mm and 5 mm thicknesses. PDO threads were also inserted. All materials were injected at a depth of 2 cm. A thermometer was used to measure heat penetration, and a multi-wavelength laser was applied to the tissue. The temperature was maintained at 60°C for 5 min to assess whether heat penetrated more than 3 cm in thickness. Observations were made regarding the heat distribution and any physical changes in the fillers and threads. In thick layers, heat accumulated above the PCL filler without penetrating deeper layers. In thin layers, heat penetration was observed. For the HA fillers, heat energy was not blocked, regardless of the G' value or thickness. For the threads, no significant heat blockage effect was observed. For all materials, no visual changes were detected in any of the materials due to temperature exposure. The findings suggest that the thickness and composition of fillers significantly influence heat penetration. Thick PCL fillers act as a thermal barrier, whereas thin PCL fillers allow deeper heat penetration. Hyaluronic acid fillers do not impede heat transfer, regardless of their physical properties. PDO threads do not exhibit any notable thermal resistance. These insights are essential for optimizing the safety and efficacy of combined filler and energy-based device treatments in esthetic medicine.

16S rRNA, metagenomics and 2bRAD-M sequencing to decode human thanatomicrobiome

Microorganisms are essential in the decomposition of corpses and play a significant role in forensic science. However, previous studies have primarily focused on animal remains, specifically the gut, skin, and burial environment. Insufficient research has been conducted on the microbiota of human cadavers, especially in cases of advanced decomposition and additional tissues, resulting in a lack of relevant reference data. In this study, the microbiota of eight cadavers at different stages of decomposition were detected using 16S rRNA, metagenomic sequencing and 2bRAD-M sequencing. Nine different sites, including oral and nasal cavities, heart, liver, spleen, lung, kidney, muscle and gut, were analysed and the efficacy of these methods was evaluated. The results showed that 16S rRNA sequencing was the most cost-effective method for the study of cadavers in the early stages of decomposition, whereas for cadaveric tissues in the late stages of decomposition, 2bRAD-M could overcome host contamination more effectively than metagenomic sequencing. This paves the way for new opportunities in data retrieval and promotes in-depth investigations into the microbiota.

Comparative Toxicological Analyses of Traditional Matrices and Blow Fly Larvae in Four Cases of Highly Decomposed Human Cadavers.

In forensic investigation, determining the time and cause of death becomes challenging, especially in cases where the remains are found in advanced decomposition, rendering traditional toxicological samples unavailable or unreliable. Entomotoxicology, an emerging methodology within forensic science, leverages insect specimens collected from cadavers as alternative toxicological samples. Several laboratory and field research studies have highlighted the efficacy in detecting various drugs, toxins, and elements absorbed by insects feeding on cadaveric tissues, even at low concentrations. However, correlation studies between drug concentrations in conventional matrices and insects remain controversial due to unknown factors influencing drug metabolism and larval feeding activity. This paper presents four real cases in which human cadavers were discovered in advanced stages of decomposition, and toxicological analyses were performed on both insect samples and available matrices. The results presented complement the scant literature currently available on the application of entomotoxicology in real cases, providing insights into the correlation between larvae and human specimen results. Furthermore, guidelines to collect and preserve entomological evidence at the crime scene and during the autopsy for use in entomotoxicological analyses are provided. This advancement holds promise in aiding forensic investigations, particularly in cases where traditional methods cannot be applied or require supporting data for further validation.

Poster 352: Histopathological Etiology of Mucoid Degeneration of the Anterior Cruciate Ligament

Objectives: Mucoid degeneration of the anterior cruciate ligament (ACL) is an uncommon clinical condition with reported prevalence below 0.5%. There is a paucity of histopathological description in the literature, which has limited our understanding of the underlying pathophysiology. Such limited understanding has resulted in treatment options limited to mechanical debridement rather than biologic interventions to prevent development or progression of mucoid degeneration. The purpose of this study is to define the histopathology of mucoid degeneration of the ACL to better guide future biologic and surgical management options. Methods: This is a case control study where a retrospective series of 10 patients were identified to have mucoid degeneration of the ACL from 2002 to 2023. They were evaluated and treated by two senior attending orthopedic sports medicine surgeons. Magnetic resonance images (MRI) were evaluated for multiple anatomic and morphometric measures. Tissue samples were obtained at the time of arthroscopic partial debridement of the ACL for all patients. Intra-operative ACL samples and cadaveric control tissues were prepared for histologic examination of microstructure and composition, and gene expression using multiplex transcriptional profiling. For the gene expression analysis RNA was isolated from archived formalin-fixed and paraffin-embedded (FFPE) sections. Three consecutive 6mm sections were pooled to generate one RNA sample and total RNA was extracted using a High Pure FFPET RNA Isolation Kit per the manufacturer’s protocol and stored at -80°C until further analysis. RNA concentration and integrity was evaluated using an Agilent BioAnalyzer 2100 and a corrected input of 50ng of total RNA was used for NanoString analyses using the NanoString nCounter® Human Fibrosis V2 Panel for multiplex molecular analysis of 770 genes. Analysis was performed using nSolver 4.0 Software and only probes with counts above background were included following the manufacturer instructions. Results: All 10 patients reported pain and limited knee flexion. MRI exhibited a bulbous appearance of the ACL. There was an increased posterior tibial slope (PTS), narrow notch width index, and increased signal intensity of the ACL. Medial meniscus tear and cartilage degeneration were reported with high prevalence. The histological analysis showed significant differences in the architecture of the tissue compared with normal ACL controls, with disorganized collagen matrix and increased glycosaminoglycan content. NanoString analysis demonstrated a total of 155 differentially expressed genes (DEGs) between the mucoid degeneration and control groups after adjustments for multiple comparisons (p&lt;0.05), with the top 20 DEGs summarized in Table 1. Among all the DEGs identified the 5 most upregulated ones include Fibronectin 1 (FN1), COL5A1, COL6A3, COL3A1 and COL1A2. There were also significant differences in the pathway scores for epithelial-to-mesenchymal transition (EMT), Extracellular matrix Degradation and Synthesis, Collagen Biosynthesis, Focal Adhesion Kinase, PDGF Signaling and PI3K-Akt (Fig. 1). The histology findings demonstrate distinct changes in the structure and composition of the ACL, and we hypothesize that repetitive microtrauma of the ACL may lead to cumulative damage that ultimately results in mucoid degeneration. Increased posterior tibial slope and narrow notch width index may contribute to cumulative loading of the ligament and subsequent mucoid degeneration. Identification of biomechanical risk factors may help to identify high risk patients. Associated injury of the medial meniscus and medial compartment cartilage also suggests that mucoid degeneration of the ACL may be part of an early degenerative process in the knee. Conclusions: Mucoid degeneration of the ACL is an uncommon condition that may represent the result of repetitive microtrauma to the ligament. Current interventions address the condition after it has progressed to an irreversible biological state. By defining the cellular and molecular mechanism(s) of the condition, we aim to broaden treatment options to potentially identify biologic interventions. The association of mucoid degeneration and medial compartment osteoarthritis suggests that patients may benefit from earlier interventions. [Figure: see text][Figure: see text]

Simultaneous multi-targeted forensic toxicological screening in biological matrices by MRM-IDA-EPI mode

The toxicologist ascertains drug assumptions in case of paediatric intoxications and death for overdose. The analytical approach consists of initially screening and consequently confirming drug positivity. We developed a toxicological screening method and validated its use comparing the results with a LC–MS/MS analysis. The method identifies 751 drugs and metabolites (704 in positive and 47 in negative mode). Chromatographic separation was achieved eluting mobile phase A (10 mM ammonium formate) and B (0.05% formic acid in methanol) in gradient on Kinetex Phenyl-Hexyl (50 × 4.6 mm, 2.6 μm) with 0.7 mL/min flow rate for 11 min. Multiple Reaction Monitoring (MRM) was adopted as survey scan and, after an Information-Dependent Analysis (IDA) (threshold of 30,000 for positive and 1000 cps for negative mode), the Enhanced Product Ion (scan range: 50–700 amu) was triggered. The MS/MS spectrum generated was compared with one of the libraries for identification. Data processing was optimised through creation of rules. Sample preparation, mainly consisting of deproteinization and enzymatic hydrolysis, was set up for different matrices (blood, urine, vitreous humor, synovial fluid, cadaveric tissues and larvae). Cut-off for most analytes resulted in the lowest concentration tested. When the results from the screening and LC–MS/MS analysis were compared, an optimal percentage of agreement (100%) was assessed for all matrices. Method applicability was evaluated on real paediatric intoxications and forensic cases. In conclusion, we proposed a multi-targeted, fast, sensitive and specific MRM-IDA-EPI screening having an extensive use in different toxicological fields.

A study protocol to characterise pathophysiological and molecular markers of rheumatic heart disease and degenerative aortic stenosis using multiparametric cardiovascular imaging and multiomics techniques.

Rheumatic heart disease (RHD), degenerative aortic stenosis (AS), and congenital valve diseases are prevalent in sub-Saharan Africa. Many knowledge gaps remain in understanding disease mechanisms, stratifying phenotypes, and prognostication. Therefore, we aimed to characterise patients through clinical profiling, imaging, histology, and molecular biomarkers to improve our understanding of the pathophysiology, diagnosis, and prognosis of RHD and AS. In this cross-sectional, case-controlled study, we plan to recruit RHD and AS patients and compare them to matched controls. Living participants will undergo clinical assessment, echocardiography, CMR and blood sampling for circulatory biomarker analyses. Tissue samples will be obtained from patients undergoing valve replacement, while healthy tissues will be obtained from cadavers. Immunohistology, proteomics, metabolomics, and transcriptome analyses will be used to analyse circulatory- and tissue-specific biomarkers. Univariate and multivariate statistical analyses will be used for hypothesis testing and identification of important biomarkers. In summary, this study aims to delineate the pathophysiology of RHD and degenerative AS using multiparametric CMR imaging. In addition to discover novel biomarkers and explore the pathomechanisms associated with RHD and AS through high-throughput profiling of the tissue and blood proteome and metabolome and provide a proof of concept of the suitability of using cadaveric tissues as controls for cardiovascular disease studies.

Surgical training in extraperitoneal laparoscopic para‑aortic lymphadenectomy for the treatment of gynecological cancer using a Thiel‑embalmed cadaver.

The extraperitoneal laparoscopic approach (ELPAN) for para-aortic lymphadenectomy provides excellent visibility of the left side of the aorta, thus facilitating surgery in the retroperitoneal space. This technique is highly complex compared with the transperitoneal approach. In particular, advanced techniques are required to develop an appropriate surgical field in the narrow retroperitoneal space; therefore, surgeons need to undergo a significant amount of training to become competent. A variety of tools are available for surgical training but are limited by their ability to reproduce complex anatomy. Thus, cadavers may represent the most suitable tool for learning this unique technique. The present study describes a surgical training protocol for the ELPAN technique using a Thiel-embalmed human cadaver and provides a step-by-step description of the ELPAN technique performed at Okayama University (Okayama, Japan). A 72-year-old Thiel-embalmed female cadaver was used to develop a protocol for surgical training in the ELPAN technique that effectively reproduced the methodology required in clinical practice. A training method for ELPAN surgery was developed and successfully completed using the Thiel-embalmed cadaver that secured the surgical field in the retroperitoneal space and permitted resection of the lymph nodes. The Thiel-embalmed cadaver tissue possessed excellent properties for surgical training, including color tone, flexibility, and the membrane structure of connective and fat tissues. In addition, this method of fixation preserved stiffness and elasticity of the peritoneum, although large vessels were slightly fragile and poorly extensible. Surgical training using a Thiel-embalmed human cadaver represents a valuable option for learning the ELPAN surgical technique. However, this technique may be unsuitable for training in perivenous manipulation. To the best of our knowledge, this is the first report to describe the use of Thiel-embalmed cadavers as a tool for surgeons to undergo training in the ELPAN technique.

Compression cycling of 3D-printed meniscal tissues in vitro using a custom bioreactor

An estimated 750,000 arthroscopic knee operations are performed in the United States each year, and many are due to a torn meniscus. Transplantation with donor tissue is the gold standard of care in cases where the meniscus cannot be repaired. However, there is a limited supply of transplantable tissue, which may not be the ideal size or shape for the recipient. 3D printing and tissue engineering have been used to produce replacement tissue of specified shape and size, but none offer the compressive modulus or durability of adult-derived tissue. While biomechanical loading of engineered tissues is known to increase mechanical strength, no current paradigms provide sufficient strength. Instead, a combinatorial approach addressing both physiological form and function has emerged as a promising strategy. In this work, anisotropic menisci were bioprinted using ink composed of collagen types I & II, chondroitin sulfate, and mesenchymal stem cells. After printing, a custom bioreactor was used to apply cyclic compression within an incubator throughout the culture period. Compression cycled prints containing cells maintained viability for 3 weeks, while the mechanical strength of cellularized prints increased after 1 week. However, print dimensions and mass of cellular prints decreased over time independent of compression, while glycosaminoglycans were lost from the prints into the culture media. The expression of eight genes were significantly altered due to compression cycling. This work demonstrated that bioprinted menisci containing live cells can be successfully compressed over long time periods in culture without cell death, and despite changing print dimensions, cells under compression contributed to meniscal strengthening whereas acellular prints consistently weaken. By optimizing structure, culture conditions, and compression paradigms, the strength of bioprinted menisci may approach that of native tissue, and this combinatorial approach may reduce or eliminate the need for cadaveric tissues for allograft transplants.

A Untethered Magnetic Micro-Robot for Local Therapeutic Delivery for Neurosurgical Oncology

INTRODUCTION: Microrobots designed for biomedical applications have garnered significant research interest, offering advantages in drug delivery, microsurgery, and biopsies. Magnetic fields enabled by electromagnetic coils permit untethered nonlinear movement of these tiny robots within the brain on the surface, the cerebrospinal fluid, and parenchyma. Microdrillers (either individually or in swarms) are a promising tool for microsurgery, capable of drilling through viscoelastic media (brain tissue). However, challenges remain in scaling down the robot size and exploring diverse applications. METHODS: Parallelizable (batch) microfabrication involved 3D printing via two-photon lithography. The pH-sensitive hydrogel was synthesized through UV-initiated radical polymerization mixed with a tracer (fluorescein sodium salt). Imaging via photon-counting CT was performed for microbot tracking in preclinical models including hydrogel phantoms, large animal cadaveric fresh brain tissue, and human cadaveric tissue. RESULTS: Microbot speeds in various tissues were characterized with speeds up to 800 um/sec over magnetic field amplitudes of 5 to 55 mT while performing complex nonlinear trajectories via open-loop surgeon control via surgeon. Targeted hydrogel delivery of fluorescein label/nanoparticle drug over a pH range of 6.2 (tumor tissue) and 7.8 (normal tissue) over a time interval of 50 s is demonstrated as proof of concept with a rate of 2.4 times faster in acidic conditions. Lastly, we show state-of-the-art micromillmetric resolution for real time imaging via photon-counting spectral CT for implementation of closed loop control. CONCLUSIONS: A novel minimally invasive microrobotic platform technology capable of targeted local drug delivery in brain parenchyma is demonstrated.

Single nuclei transcriptomics of the in situ human limbal stem cell niche

The corneal epithelium acts as a barrier to pathogens entering the eye; corneal epithelial cells are continuously renewed by uni-potent, quiescent limbal stem cells (LSCs) located at the limbus, where the cornea transitions to conjunctiva. There has yet to be a consensus on LSC markers and their transcriptome profile is not fully understood, which may be due to using cadaveric tissue without an intact stem cell niche for transcriptomics. In this study, we addressed this problem by using single nuclei RNA sequencing (snRNAseq) on healthy human limbal tissue that was immediately snap-frozen after excision from patients undergoing cataract surgery. We identified the quiescent LSCs as a sub-population of corneal epithelial cells with a low level of total transcript counts. Moreover, TP63, KRT15, CXCL14, and ITGβ4 were found to be highly expressed in LSCs and transiently amplifying cells (TACs), which constitute the corneal epithelial progenitor populations at the limbus. The surface markers SLC6A6 and ITGβ4 could be used to enrich human corneal epithelial cell progenitors, which were also found to specifically express the putative limbal progenitor cell markers MMP10 and AC093496.1.

Quality and applicability of cadaveric donor eyes for molecular biology research: An Indian experience.

Human ocular tissue banking plays an important part in the advancement of translational research for identifying the molecular processes involved in disease etiology and pathogenesis. Timely obtaining a good-quality ocular tissue from a cadaveric donor is exceedingly difficult, especially in remote areas, with a variable transportation time (within 12-24 h), raising concerns about RNA quality and its subsequent applications. Therefore, we assessed the utility of retinal tissues from cadaver donor and enucleated eyes based on the RNA quality and gene expression by real-time polymerase chain reaction (PCR). Prospective study. Retina tissues were separated from the donor/enucleated eyes received in the eye bank within 24 h of death (n = 15) and within an hour from OR (n = 3), respectively, and stored immediately at -80 degree. RNA was isolated using trizol, and the quantity and quality were assessed using Qubit and agarose gel electrophoresis, respectively. QPCR was performed for measuring the expression of different retinal-specific genes. The cellular viability of the retina was assessed by establishing explant primary cell cultures. The data were calculated as an average of normalised Ct values ± standard error of the mean. RNA obtained from cadaveric tissues despite being partially degraded showed a uniform strong gene expression of several retinal-specific genes such as PAX6, RHO, TUBB3, CRX , and ALDH1L1 . The primary cultures established from cadaveric tissues showed viable cells. The cadaver donor tissues collected within 24 hours of death can be effectively utilized for gene expression profiling.

Virtual histological staining of unlabeled autopsy tissue

Traditional histochemical staining of post-mortem samples often confronts inferior staining quality due to autolysis caused by delayed fixation of cadaver tissue, and such chemical staining procedures covering large tissue areas demand substantial labor, cost and time. Here, we demonstrate virtual staining of autopsy tissue using a trained neural network to rapidly transform autofluorescence images of label-free autopsy tissue sections into brightfield equivalent images, matching hematoxylin and eosin (H&E) stained versions of the same samples. The trained model can effectively accentuate nuclear, cytoplasmic and extracellular features in new autopsy tissue samples that experienced severe autolysis, such as COVID-19 samples never seen before, where the traditional histochemical staining fails to provide consistent staining quality. This virtual autopsy staining technique provides a rapid and resource-efficient solution to generate artifact-free H&E stains despite severe autolysis and cell death, also reducing labor, cost and infrastructure requirements associated with the standard histochemical staining.