Development of new nanofibrous nerve conduits by PCL-Chitosan-Hyaluronic acid containing Piracetam-Vitamin B12 for sciatic nerve: A rat model

• Published in: International journal of pharmaceutics Vol. 655; p. 123978
• Main Authors: Jafarisavari, Zahra, Ai, Jafar, Abbas Mirzaei, Seyed, Soleimannejad, Mostafa, Asadpour, Shiva
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 25.04.2024
• Subjects: Animals; Chitosan; Hyaluronic Acid; Nanofiber; Nanofibers; Nerve Guidance Conduits; Nerve Regeneration; PC12 Cells; Peripheral Nerve Injuries - drug therapy; Peripheral Nerve Injuries - pathology; Piracetam; Polycaprolactone; Rats; Rats, Wistar; Sciatic Nerve; Tissue Scaffolds; Vitamin B 12; Vitamin B12; Nerve Guidance Conduits; Hyaluronic Acid; Nanofiber; Piracetam; Chitosan; Sciatic Nerve; Vitamin B12; Polycaprolactone
• ISSN: 0378-5173; 1873-3476
• DOI: 10.1016/j.ijpharm.2024.123978

Schematic image of conduit fabrication (sheets and tubular nerve conduits) by electrospinning technique; In vitro conduit characterization tests: Scanning electron microscopy (SEM), Attenuated total reflectance (ATR)-Fourier Transforms Infrared (FTIR) spectroscopy, Surface hydrophilicity test (Contact angle), Mechanical tests, 4′,6-diamidino-2-phenylindole (DAPI); In vivo study: Implantation of nanofibrous tubular conduits in a rat model with sciatic nerve defects (B); Behavioral functional test with walking foot print test to measure sciatic nerve index (SFI), Histopathological staining with hematoxylin and eosin (H&E) and Anti-Neurofilament 200 (NF200). CH: Chitosan, PIR: Piracetam, VIT B12: Vitamin B12, PCL: Polycaprolactone, SEM: Scanning electron microscopy, ATR-FTIR: Attenuated total reflectance (ATR)-Fourier Transforms Infrared (FTIR) spectroscopy, DAPI: 4′,6-diamidino-2-phenylindole, H&E: hematoxylin and eosin, NF200: Anti-Neurofilament 200. [Display omitted] •The flexibility of the PCL/CH/PIR/VITB12 conduit (29.78 ± 0.69 %) was close to human sciatic nerve (30.2 ± 3.1).•The PCL/CH/PIR/VITB12 conduit had a sufficient resistance to collapse, and tear during implantation.•The nerve conduits promoted myelination, axon regeneration, and functional recovery in rat sciatic nerve defect model. Peripheral nerve injury is a critical condition that can disrupt nerve functions. Despite the progress in engineering artificial nerve guidance conduits (NGCs), nerve regeneration remains challenging. Here, we developed new nanofibrous NGCs using polycaprolactone (PCL) and chitosan (CH) containing piracetam (PIR)/vitamin B12(VITB12) with an electrospinning method. The lumen of NGCs was coated by hyaluronic acid (HA) to promote regeneration in sciatic nerve injury. The NGCs were characterized via Scanning Electron Microscopy (SEM), Fourier transform infrared (FTIR), tensile, swelling, contact angle, degradation, and drug release tests. Neuronal precursor cell line (PCL12 cell) and rat mesenchymal stem cells derived from bone marrow (MSCs) were seeded on the nanofibrous conduits. After that, the biocompatibility of the NGCs was evaluated by the 2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, 4′,6-diamidino-2-phenylindole (DAPI) staining, and SEM images. The SEM demonstrated that PCL/CH/PIR/VITB12 NGCs had nonaligned, interconnected, smooth fibers. The mechanical properties of these NGCs were similar to rat sciatic nerve. These conduits had an appropriate swelling and degradation rate. The In Vitro studies exhibited favorable biocompatibility of the PCL/CH/PIR/VITB12 NGCs towards PC12 cells and MSCs. The in vitro studies exhibited favorable biocompatibility of the PCL/CH/PIR/VIT B12 NGCs towards MSCs and PC12 cells. To analyze functional efficacy, NGCs were implanted into a 10 mm Wistar rat sciatic nerve gap and bridged the proximal and distal stump of the defect. After three months, the results of sciatic functional index (55.3 ± 1.8), hot plate latency test (5.6 ± 0.5 s), gastrocnemius muscle wet weight-loss (38.57 ± 1.6 %) and histopathological examination using hematoxylin-eosin (H&E) /toluidine blue/ Anti-Neurofilament (NF200) staining demonstrated that the produced conduit recovered motor and sensory functions and had comparable nerve regeneration compared to the autograft that can be as the gold standard to bridge the nerve gaps.