Toward optoacoustic sciatic nerve detection using an all-fiber interferometric-based sensor for endoscopic smart laser surgery.

OBJECTIVES: Laser surgery requires efficient tissue classification to reduce the probability of undesirable or unwanted tissue damage. This study aimed to investigate acoustic shock waves (ASWs) as a means of classifying sciatic nerve tissue. MATERIALS AND METHODS: In this study, we classified sciatic nerve tissue against other tissue types-hard bone, soft bone, fat, muscle, and skin extracted from two proximal and distal fresh porcine femurs-using the ASWs generated by a laser during ablation. A nanosecond frequency-doubled Nd:YAG laser at 532 nm was used to create 10 craters on each tissue type's surface. We used a fiber-coupled Fabry-Pérot sensor to measure the ASWs. The spectrum's amplitude from each ASW frequency band measured was used as input for principal component analysis (PCA). PCA was combined with an artificial neural network to classify the tissue types. A confusion matrix and receiver operating characteristic (ROC) analysis was used to calculate the accuracy of the testing-data-based scores from the sciatic nerve and the area under the ROC curve (AUC) with a 95% confidence-level interval. RESULTS: Based on the confusion matrix and ROC analysis of the model's tissue classification results (leave-one-out cross-validation), nerve tissue could be classified with an average accuracy rate and AUC result of 95.78  ± 1.3% and 99.58  ± 0.6%, respectively. CONCLUSION: This study demonstrates the potential of using ASWs for remote classification of nerve and other tissue types. The technique can serve as the basis of a feedback control system to detect and preserve sciatic nerves in endoscopic laser surgery.

Mouse and human HSPC immobilization in liquid culture by CD43- or CD44-antibody coating.

Keeping track of individual cell identifications is imperative to the study of dynamic single-cell behavior over time. Highly motile hematopoietic stem and progenitor cells (HSPCs) migrate quickly and do not adhere, and thus must be imaged very frequently to keep cell identifications. Even worse, they are also flushed away during medium exchange. To overcome these limitations, we tested antibody coating for reducing HSPC motility in vitro. Anti-CD43- and anti-CD44-antibody coating reduced the cell motility of mouse and human HSPCs in a concentration-dependent manner. This enables 2-dimensional (2D) colony formation without cell mixing in liquid cultures, massively increases time-lapse imaging throughput, and also maintains cell positions during media exchange. Anti-CD43 but not anti-CD44 coating reduces mouse HSPC proliferation with increasing concentrations. No relevant effects on cell survival or myeloid and megakaryocyte differentiation of hematopoietic stem cells and multipotent progenitors 1-5 were detected. Human umbilical cord hematopoietic CD34+ cell survival, proliferation, and differentiation were not affected by either coating. This approach both massively simplifies and accelerates continuous analysis of suspension cells, and enables the study of their behavior in dynamic rather than static culture conditions over time.

Optimized Decellularization Protocol for Large Peripheral Nerve Segments: Towards Personalized Nerve Bioengineering.

Nerve injuries remain clinically challenging, and allografts showed great promise. Decellularized nerve allografts possess excellent biocompatibility and biological activity. However, the vast majority of decellularization protocols were established for small-size rodent nerves and are not suitable for clinical application. We aimed at developing a new method of decellularizing large-diameter nerves suitable for human transplantation. Repeated rounds of optimization to remove immunogenic material and preserve the extracellular structure were applied to the porcine sciatic nerve. Following optimization, extensive in vitro analysis of the acellular grafts via immunocytochemistry, immunohistology, proteomics and cell transplantation studies were performed. Large segments (up to 8 cm) of the porcine sciatic nerve were efficiently decellularized and histology, microscopy and proteomics analysis showed sufficient preservation of the extracellular matrix, with simultaneous consistent removal of immunogenic material such as myelin, DNA and axons, and axonal growth inhibitory molecules. Cell studies also demonstrated the suitability of these acellular grafts for 3D cell culture studies and translation to future large animal studies and clinical trials. By using non-human donors for peripheral nerve transplantation, significant drawbacks associated with the gold standard can be eliminated while simultaneously preserving the beneficial features of the extracellular matrix.

Schwann Cell-Like Cells: Origin and Usability for Repair and Regeneration of the Peripheral and Central Nervous System.

Functional recovery after neurotmesis, a complete transection of the nerve fiber, is often poor and requires a surgical procedure. Especially for longer gaps (>3 mm), end-to-end suturing of the proximal to the distal part is not possible, thus requiring nerve graft implantation. Artificial nerve grafts, i.e., hollow fibers, hydrogels, chitosan, collagen conduits, and decellularized scaffolds hold promise provided that these structures are populated with Schwann cells (SC) that are widely accepted to promote peripheral and spinal cord regeneration. However, these cells must be collected from the healthy peripheral nerves, resulting in significant time delay for treatment and undesired morbidities for the donors. Therefore, there is a clear need to explore the viable source of cells with a regenerative potential similar to SC. For this, we analyzed the literature for the generation of Schwann cell-like cells (SCLC) from stem cells of different origins (i.e., mesenchymal stem cells, pluripotent stem cells, and genetically programmed somatic cells) and compared their biological performance to promote axonal regeneration. Thus, the present review accounts for current developments in the field of SCLC differentiation, their applications in peripheral and central nervous system injury, and provides insights for future strategies.