Numerous studies in animals have shown the unique property of olfactory ensheathing cells to stimulate regeneration of lesioned axons in the spinal cord. In a Phase I clinical trial, we assessed the safety and feasibility of transplantation of autologous mucosal olfactory ensheathing cells and olfactory nerve fibroblasts in patients with complete spinal cord injury. Six patients with chronic thoracic paraplegia (American Spinal Injury Association class A-ASIA A) were enrolled for the study. Three patients were operated, and three served as a control group. The trial protocol consisted of pre-and postoperative neurorehabilitation, olfactory mucosal biopsy, culture of olfactory ensheathing cells, and intraspinal cell grafting. Patient's clinical state was evaluated by clinical, neurophysiological, and radiological tests. There were no adverse findings related to olfactory mucosa biopsy or transplantation of olfactory ensheathing cells at 1 year after surgery. There was no evidence of neurological deterioration, neuropathic pain, neuroinfection, or tumorigenesis. In one cell-grafted patient, an asymptomatic syringomyelia was observed. Neurological improvement was observed only in transplant recipients. The first two operated patients improved from ASIA A to ASIA C and ASIA B. Diffusion tensor imaging showed restitution of continuity of some white matter tracts throughout the focus of spinal cord injury in these patients. The third operated patient, although remaining ASIA A, showed improved motor and sensory function of the first spinal cords segments below the level of injury. Neurophysiological examinations showed improvement in spinal cord transmission and activity of lower extremity muscles in surgically treated patients but not in patients receiving only neurorehabilitation. Observations at 1 year indicate that the obtaining, culture, and intraspinal transplantation of autologous olfactory ensheathing cells were safe and feasible. The significance of the neurological improvement in the transplant recipients and the extent to which the cell transplants contributed to it will require larger numbers of patients.

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Research concerning the elaboration and application of biomaterial which may support the nerve tissue regeneration is currently one of the most promising directions. Biocompatible polymer devices are noteworthy group among the numerous types of potentially attractive biomaterials for regenerative medicine application. Polylactides and polyurethanes may be utilized for developing devices for supporting the nerve regeneration, like nerve guide conduits or bridges connecting the endings of broken nerve tracts. Moreover, the combination of these biomaterial devices with regenerative cell populations, like stem or precursor cells should significantly improve the final therapeutic effect. Therefore, the composition and structure of final device should support the proper adhesion and growth of cells destined for clinical application. In current research, the three polymer mats elaborated for connecting the broken nerve tracts, made from polylactide, polyurethane and their blend were evaluated both for physical properties and in vitro, using the olfactory-bulb glial cells and mesenchymal stem cells. The evaluation of Young's modulus, wettability and roughness of obtained materials showed the differences between analyzed samples. The analysis of cell adhesion, proliferation and morphology showed that the polyurethane-polylactide blend was the most neutral for cells in culture, while in the pure polymer samples there were significant alterations observed. Our results indicated that polyurethane-polylactide blend is an optimal composition for culturing and delivery of glial and mesenchymal stem cells.

Recently, we described the influence of sodium alginate on the inflammatory infiltrate during neuroregeneration in tube nerve grafts. It was noticeable that there was the coexistence of inflammatory cells, including neutrophils, plasma cells, and macrophages with Schwann cells and axons. This may indicate a beneficial interaction between alginates and the infiltrate and the additional beneficial effect of the cells on the neuroregeneration process in the inflammatory infiltrates. In this study, we have performed in vivo evaluation of our novel tubular implant prepared by a polyurethane/polylactide blend filled with alginate fibers. The influence of filling the lumen of the tubes with sodium and calcium alginates on the regeneration process of the rat sciatic nerve was investigated. The neuroregeneration process was assessed by detailed histomorphometric studies, axon counting, and calculating the regenerative indexes. It was observed that calcium alginate had a supportive effect on nerve regeneration similar to nerve autotransplant.

Spinal cord injury (SCI) is a well-known devastating lesion that sadly is very resistant to all treatment attempts. This fact has stimulated the exploration of multiple regenerative strategies that are examined at both the basic and clinical level. For laboratory research, different in vivo models are used, but each has many important limitations. The main limitation of these models is the high level of animal suffering related to the inflicted neurologic injury. It has caused a growing tendency to limit the injury, but this, in turn, produces incomplete SCI models and uncertainties in the neuroregeneration interpretation. To overcome such limitations, a new experimental SCI model is proposed. Geckos have been extensively examined as a potential animal model of SCI. Their spinal cord extends into the tail and can be transected without causing the typical neurologic consequences observed in rat models. In this study, we compared the gecko tail SCI model with the rat model of thoracic SCI. Anatomic and histologic analyses showed comparability between the gecko and rat in diameter of spinal canal and spinal cord, as well as applicability of multiple staining techniques (hematoxylin and eosin, immunostaining, and scanning and transmission electron microscopy). We tested the suitability of in vivo study with 3 prototype implants for the reconstruction of SCI: a multichannel sponge, a multilaminar tube, and a gel cylinder. These were compared with a spinal cord excision (control). A 20-wk observation revealed no adverse effects of SCI on the animals' well-being. The animals were easily housed and observed. Histologic analysis showed growth of nervous tissue elements on implant surface and implant cellular colonization. The study showed that the gecko SCI model can be used as a primary model for the assessment of SCI treatment methods. It provides a platform for testing multiple solutions with limited animal suffering before performing tests on mammals. Detailed results of the experimental conditions and testing techniques are provided.-Szarek, D., Marycz, K., Lis, A., Zawada, Z., Tabakow, P., Laska, J., Jarmundowicz, W. Spinal cord injury (SCI) is a lesion that has a relatively high incidence (1) and is characterized by high resistance to any restorative treatment. Multiple experimental, laboratory, and clinical investigations are conducted in an attempt to understand the mechanisms underlying this trauma and to identify potential treatment options. Studies are performed in in vitro and in vivo models that have both pros and cons. In vitro studies are relatively easy to perform and repeat; they do not require animal euthanasia and can be performed in large numbers. The disadvantage of this type of study is a relatively short observation time and examination limited to 1 or 2 different cell lines during 1 test. Moreover, they do not provide data on how the whole organism may respond to the study drug, implant, or experimental condition. On the other hand, in vivo studies provide information on how multiple tiss...

This preliminary research project has been conducted to evaluate different elastic polymer materials in terms of their applicability in peripheral nerve regeneration. Poly(tetrafluoroetylene-co-difluorovinylidene-co-propylene), poly(L-lactide-co-D,L-lactide), and polyurethane were used for the manufacture of tubular implants. Alginate sodium gel and fibers were used as a scaffold to fill in tube nerve grafts and enhance nerve regeneration. The tubes were implanted to reconstruct a 10 mm gap in the sciatic nerve in rats. After 3, 7, 14, 28 days the tubes were retrieved for histological examination. Among tested tubes polyurethane implants were found to be the most suitable because of their mechanical and surgical properties. Other tested implants were found to be unfavorable due to their inappropriate rigidity, elasticity or surgical convenience. Alginate transformation into dense gel form was observed that hindered inner tube space cellular colonization. In consequence of this transformation nerve regeneration was inhibited inside tube nerve grafts. Histological examination showed massive colonization of the implants with Schwann cells, and growth of new axons was found within Schwann cells growing on tubes external surface. Appropriate time rates for alginate gelation and dissolving must be determined to allow undisturbed tissue growth and maturation

The aim of this research was to evaluate novel biomaterials for neural regeneration. The investigated materials were composed of polyurethane (PU) and polylactide (PLDL) blended at three different w/w ratios, that is, 5/5, 6/4, and 8/2 of PU/PLDL. Ultrathin fibrous scaffolds were prepared using electrospinning. The scaffolds were investigated for their applicability for nerve regeneration by culturing rat olfactory ensheathing glial cells. Cells were cultured on the materials for seven days, during which cellular morphology, phenotype, and metabolic activity were analysed. SEM analysis of the fabricated fibrous scaffolds showed fibers of a diameter mainly lower than 600 m with unimportant volume of protrusions situated along the fibers, with nonsignificant differences between all analysed materials. Cells cultured on the materials showed differences in their morphology and metabolic activity, depending on the blend composition. The most proper morphology, with numerous p75 + and GFAP + cells present, was observed in the sample 6/4, whereas the highest metabolic activity was measured in the sample 5/5. However, none of the investigated samples showed cytotoxicity or negatively influenced cellular morphology. Therefore, the novel electrospun fibrous materials may be considered for regenerative medicine applications, and especially when contacting with highly sensitive nervous cells.

Traumatic brain injury (TBI) is a major public health problem among children. The predominant causes of TBI in young children are motor vehicle accidents, firearm incidents, falls, and child abuse. The limitation of in vivo studies on the human brain has made the finite element modelling an important tool to study brain injury. Numerical models based on the finite element approach can provide valuable data on biomechanics of brain tissues and help explain many pathological conditions. This work reviews the existing numerical models of a child’s head. However, the existing literature is very limited in reporting proper geometric representation of a small child’s head. Therefore, an advanced 2-year-old child’s head model, named aHEAD 2yo (aHEAD: advanced Head models for safety Enhancement And medical Development), has been developed, which advances the state-of-the-art. The model is one of the first published in the literature, which entirely consists of hexahedral elements for three-dimensional (3D) structures of the head, such as the cerebellum, skull, and cerebrum with detailed geometry of gyri and sulci. It includes cerebrospinal fluid as Smoothed Particle Hydrodynamics (SPH) and a detailed model of pressurized bringing veins. Moreover, the presented review of the literature showed that material models for children are now one of the major limitations. There is also no unambiguous opinion as to the use of separate materials for gray and white matter. Thus, this work examines the impact of various material models for the brain on the biomechanical response of the brain tissues during the mechanical loading described by Hardy et al. The study compares the inhomogeneous models with the separation of gray and white matter against the homogeneous models, i.e., without the gray/white matter separation. The developed model along with its verification aims to establish a further benchmark in finite element head modelling for children and can potentially provide new insights into injury mechanisms.