Liposome-exosome hybrids for in situ detection of exosomal miR-1246 in breast cancer.

Several lines of evidence suggest that exosomal miRNAs are potential biomarkers for cancer monitoring. An urgent need remains for the in situ detection of exosomal miRNAs at low concentrations without destroying the exosome structure. In the present study, a novel sensitive exosomal miR-1246 in situ detection strategy has been developed by integrating the CRISPR/Cas13a system with the formation of hybrids between exosomes and cationic liposomes. The liposomes were loaded with CRISPR/Cas13a, CRISPR RNA (crRNA), and RNA reporter probes. In the presence of exosomes, the liposome-exosome hybrids were formed through electrostatic interactions, and CRISPR/Cas13a was activated to cleave the reporter probes by exosomal miR-1246. The acquired fluorescence signal showed a linear response to the logarithm of MCF-7 exosome concentrations, indicating a quantitative response to exosomal miR-1246. The regression equation is y = 5021 log C - 9976 (R2 = 0.9985) with a limit of detection of 3 × 102 particles per mL. This strategy could not only be used to detect serum exosomal miR-1246 in breast cancer patients but also to distinguish early form advanced disease. This strategy can be exploited in future exosomal miRNA analyses.

Polyglycolic acid filaments guide Schwann cell migration in vitro and in vivo.

Nerve conduits filled with longitudinal aligned filaments have demonstrated a better regenerative outcome for bridging large peripheral nerve gaps than hollow nerve conduits. In the present study, we investigated the in vitro and in vitro cellular behavior of Schwann cells on polyglycolic acid (PGA) filaments by immunocyto/histochemistry and light/electron microscopy. After 1-3-week culture of rat dorsal root ganglia (DRGs) onto PGA filaments, Schwann cells from rat DRGs adhered to and migrated along PGA filaments. Twenty-four rats received implantation of chitosan conduits inserted with PGA filaments to bridge 10-mm-long sciatic nerve gaps. At 1, 2, 3 and 4 weeks post-implantation (n = 6, each time point), Schwann cells were found to migrate along PGA filaments and form cell columns resembling bands of Büngner. These results suggest that PGA filaments may play a contact guidance role in Schwann cell migration and thus serve as a promising conduit-filling material to facilitate peripheral nerve repair.

Repairing a 35-mm-long median nerve defect with a chitosan/PGA artificial nerve graft in the human: a case study.

We have developed a chitosan/polyglycolic acid (PGA) artificial nerve graft which was previously used for bridge implantation of dog sciatic nerves across 30-mm long defects. Here we describe a clinical trial of this graft for repairing a 35-mm-long median nerve defect at elbow of a human patient. During the 3-year follow-up period, functional recovery of the injured median nerve was assessed by pinch gauge test, hydraulic hand dynamometry, static two-point discrimination and touch test with monofilaments, in couple with electrophysiological examinations. The motor and sensory function of the median nerve demonstrated an ongoing recovery postimplantation, reaching M4 and S3+ levels during the follow-up period. The results indicate that the chitosan/PGA artificial nerve graft could be used for surgical repair of larger defects in major peripheral nerves at a higher level in the human.

Surgical repair of a 30 mm long human median nerve defect in the distal forearm by implantation of a chitosan-PGA nerve guidance conduit.

This paper describes a clinical case study in which a chitosan/polyglycolic acid nerve guidance conduit (chitosan-PGA NGC) was utilized to repair a 30 mm long median nerve defect in the right distal forearm of a 55 year-old male patient. Thirty-six months after the nerve repair, the palm abduction of the thumb and the thumb-index digital opposition recovered, facilitating the patient to accomplish fine activities, such as handling chopsticks. Static two-point discrimination measured 14, 9 and 9 mm in the thumb, index and middle fingers of the right hand. Reproducible compound muscle action potentials were recorded on the right abductor pollicis. The ninhydrin test, a classical method for assessing sympathetic nerve function, showed partial recovery of the perspiration function of the injured thumb, index and middle fingers. This repair case suggested a possible strategy for the clinical reconstruction of extended defects in human peripheral nerve trunks by the implantation of chitosan-PGA NGCs.