Chitosan Tubes Research Articles

Graphene/ chitosan tubes inoculated with dental pulp stem cells promotes repair of facial nerve injury.

Introduction: Facial nerve injury significantly impacts both the physical and psychological] wellbeing of patients. Despite advancements, there are still limitations associated with autografts transplantation. Consequently, there is an urgent need for effective artificial grafts to address these limitations and repair injuries. Recent years have witnessed the recognition of the beneficial effects of chitosan (CS) and graphene in the realm of nerve repair. Dental pulp stem cells (DPSCs) hold great promise due to their high proliferative and multi-directional differentiation capabilities. Methods: In this study, Graphene/CS (G/CST) composite tubes were synthesized and their physical, chemical and biological properties were evaluated, then DPSCs were employed as seed cells and G/CST as a scaffold to investigate their combined effect on promoting facial nerve injury repair. Results and Disscussion: The experimental results indicate that G/CST possesses favorable physical and chemical properties, along with good cyto-compatibility. making it suitable for repairing facial nerve transection injuries. Furthermore, the synergistic application of G/CST and DPSCs significantly enhanced the repair process for a 10mm facial nerve defect in rabbits, highlighting the efficacy of graphene as a reinforcement material and DPSCs as a functional material in facial nerve injury repair. This approach offers an effective treatment strategy and introduces a novel concept for clinically managing facial nerve injuries.

Inferior long-term Results of a Prospective Randomized Controlled Trial initially demonstrating enhanced Sensory Nerve Recovery using a Chitosan Nerve Tube.

Traumatic peripheral nerve injuries can result in significant functional impairments and long-term sequelae. This study evaluated the long-term outcomes of a chitosan tube implantation protecting the epineural coaptation after peripheral nerve injuries using two different tube versions (V 1.0 and V 2.0 with different wall thickness and resorption characteristics) compared to a control group. The study focused on pain levels, sensory function, and overall functional outcomes. Patients who received tube implantation around direct coaptation sites of digital nerves were prospectively randomized and compared to control patients without additional tube protection. Pain levels, sensory function, grip force, and functional scores were assessed at different time points, ranging from three months to five years after the procedure. Furthermore, biodegradation of the tubes was measured via high-resolution MR-neurography (MRN) and categorized. Long-term evaluation revealed that patients with V 1.0 had higher pain levels compared to the control group after five years. They also reported more symptoms of numbness and hypersensitivity. V 2.0 patients exhibited higher pain levels at three months, which did not persist at six months. However, they showed compromised sensory function, with higher values of two-point discrimination compared to V 1.0 and the control group. No differences were found in grip force or functional scores between the groups. MRI displayed remnants of implants even in long-term follow-up. The findings suggest potential limitations due to pain increase and impaired sensory function associated with tube implantation in the long term. However, in the short term, the material seemed to have a protective effect (as published previously). The resorption process was not completed at the end of the observation period of five years. This might explain the prolonged scarring and inferior long-term results. Future research should focus on improving tube materials and design to minimize adverse effects and enhance functional outcomes in patients with peripheral nerve injuries.

Closing the Gap: Bridging Peripheral Sensory Nerve Defects with a Chitosan-Based Conduit a Randomized Prospective Clinical Trial.

Introduction: If tensionless nerve coaptation is not possible, bridging the resulting peripheral nerve defect with an autologous nerve graft is still the current gold standard. The concept of conduits as an alternative with different materials and architectures, such as autologous vein conduits or bioartificial nerve conduits, could not replace the nerve graft until today. Chitosan, as a relatively new biomaterial, has recently demonstrated exceptional biocompatibility and material stability with neural lineage cells. The purpose of this prospective randomized clinical experiment was to determine the efficacy of chitosan-based nerve conduits in regenerating sensory nerves in the hand. Materials and methods: Forty-seven patients with peripheral nerve defects up to 26 mm distal to the carpal tunnel were randomized to receive either a chitosan conduit or an autologous nerve graft with the latter serving as the control group. Fifteen patients from the conduit group and seven patients from the control group were available for a 12-month follow-up examination. The primary outcome parameter was tactile gnosis measured with two-point discrimination. The secondary outcome parameters were Semmens Weinstein Monofilament Testing, self-assessed pain, and patient satisfaction. Results: Significant improvement (in static two-point discrimination) was observed six months after trauma (10.7 ± 1.2 mm; p < 0.05) for chitosan-based nerve conduits, but no further improvement was observed after 12 months of regeneration (10.9 ± 1.3 mm). After six months and twelve months, the autologous nerve graft demonstrated comparable results to the nerve conduit, with a static two-point discrimination of 11.0 ± 2.0 mm and 7.9 ± 1.1 mm. Semmes Weinstein Filament Testing in the nerve conduit group showed a continuous improvement over the regeneration period by reaching from 3.1 ± 0.3 after three months up to 3.7 ± 0.4 after twelve months. Autologous nerve grafts presented similar results: 3.3 ± 0.4 after three months and 3.7 ± 0.5 after twelve months. Patient satisfaction and self-reported pain levels were similar between the chitosan nerve conduit and nerve graft groups. One patient required revision surgery due to complications associated with the chitosan nerve tube. Conclusion: Chitosan-based nerve conduits are safe and suitable for bridging nerve lesions up to 26 mm in the hand. Tactile gnosis improved significantly during the early regeneration period, and functional outcomes were similar to those obtained with an autologous nerve graft. Thus, chitosan appears to be a sufficient substitute for autologous nerve grafts in the treatment of small nerve defects in the hand.

Chitosan Tubes Inoculated with Dental Pulp Stem Cells and Stem Cell Factor Enhance Facial Nerve-Vascularized Regeneration in Rabbits.

Facial nerve injury is a common clinical condition that leads to disfigurement and emotional distress in the affected individuals, and the recovery presents clinical challenges. Tissue engineering is the standard method to repair nerve defects. However, nerve regeneration is still not satisfactory because of poor neovascularization after implantation, especially for the long-segment nerve defects. In the current study, we aimed to investigate the potential of chitosan tubes inoculated with stem cell factor (SCF) and dental pulp stem cells (DPSCs) in facial nerve-vascularized regeneration. In the in vitro experiment, DPSCs were isolated, cultured, and then identified. The optimal concentration of SCF was screened by CCK8. Cytoskeleton and living-cell staining, migration, CCK8 test, and neural differentiation assays were performed, revealing that SCF promoted the biological activity of DPSCs. Surprisingly, SCF increased the neural differentiation of DPSCs. The migration and angiogenesis experiments were carried out to show that SCF promoted the angiogenesis and migration of human umbilical vein endothelial cells (HUVECs). In the facial nerve, 7 mm defects of New Zealand white rabbits, hematoxylin–eosin (HE), immunohistochemistry, toluidine blue staining, and transmission electron microscopy observation were performed at 12 weeks postsurgery to show more nerve fibers and better myelin sheath in the SCF + DPSC group. In addition, the whisker movements, Masson’s staining, and western blot assays were performed, demonstrating functional repair and that the expression level of CD31 protein in the group SCF + DPSCs was relatively close to that in the group Autograft. In summary, chitosan tubes inoculated with SCF and DPSCs increased neurovascularization and provided an effective method for repairing facial nerve defects, indicating great promise for clinical application.

Pattern selection of directionally oriented chitosan tubes.

The growth of viscoelastic curved materials, inspired by biological systems, may give rise to various complex structures. One of the simplest ways to control the pattern formation is to vary the orientation of the reaction vessel while keeping all other experimental conditions constant. Here, we report the self-organization of soft chitosan tubes by injecting acidic chitosan sol into a pool of sodium hydroxide solution, where the adhesive force between the gel and container keeps the tubules on the bottom of the reactor. The horizontal growth of the tubular structure undergoes spontaneous symmetry breaking, where instabilities develop on the surface of the chitosan tubules. Transformation of folds into wrinkles and finally to a smooth tube takes place by varying the orientation of the container. In addition to characterizing the evolving structures, we have also shown that the linear growth rate of the tube scales with the tilt angle of the container from the horizontal.

Effect of nanotubes on the electrical and mechanical properties of chitosan films

Using the methods of X-ray diffraction and scanning electron microscopy, the structure of composite films based on chitosan and single-wall carbon tubes has been studied. It is shown that the introduction of carbon nanotubes leads to the ordering of the chitosan structure. Increase in concentration of nanotubes (from 0 to 3%) causes rise in the value of storage modulus from 3 to 4 GPa (DMA data), increase in electrical conductivity of samples (from 10-11 to 102 S/m), and some changes in their dielectric permittivity (from 5.5. to 26 at an electrical field frequency of 1 kHz). Data on the ionic and electronic components of the conductivity of the composite film are presented. Keywords: chitosan, electrical conductivity, nanotubes, dielectric constant, composite.

Влияние углеродных нанотрубок на электрические и механические свойства хитозановых пленок

Using the methods of X-ray diffraction and scanning electron microscopy, the structure of composite films based on chitosan and single-wall carbon tubes has been studied. It is shown that the introduction of carbon nanotubes leads to the ordering of the chitosan structure. Increase in concentration of nanotubes (from 0 to 3%) causes rise in the value of storage modulus from 3 to 4 GPa (DMA data), increase in electrical conductivity of samples (from 10-11 to 102 S/m), and some changes in their dielectric permittivity (from 5.5. to 26 at an electrical field frequency of 1kHz). Data on the ionic and electronic components of the conductivity of the composite film are presented.

Chitosan Tubes Prefilled with Aligned Fibrin Nanofiber Hydrogel Enhance Facial Nerve Regeneration in Rabbits.

Facial nerves are fragile and easily injured, for example, by traffic accidents or operations. Facial nerve injury drastically reduces the quality of life in affected patients, and its treatment presents clinical challenges. A promising therapeutic strategy includes nerve conduits with appropriate fillers capable of guiding nerve regeneration. In this study, a three-dimensional hierarchically aligned fibrin nanofiber hydrogel (AFG) assembled via electrospinning and molecular self-assembly was first used to mimic the architecture of the native fibrin cable, which is similar to the nerve extracellular matrix (ECM). AFG as a substrate in chitosan tubes (CST) was used to bridge a 7 mm-long gap in a rabbit buccal branch facial nerve defect model. The results showed that AFG and CST showed good compatibility to support the adhesion, activity, and proliferation of Schwann cells (SCs). Further morphological, histological, and functional analyses demonstrated that the regenerative outcome of AFG-prefilled CST was close to that of autologous nerve grafts and superior to that of CST alone or CSTs prefilled with random fibrin nanofiber hydrogel (RFG), which indicated that AFG-prefilled CST markedly improved axonal regeneration with enhanced remyelination and functional recovery, thus showing great potential for clinical application for facial nerve regeneration treatments.

Tubular chitosan device for use as prosthesis coating in vascular surgery

Chitosan is a natural, biodegradable, non-toxic and biocompatible polymer, with characteristics such as a healing, hemostatic, antimicrobial agent, among others. Therefore, the aim of this study is to develop a tubular chitosan device for use as a prosthetic coating application in vascular surgery. The chitosan wires were obtained by the spinning method in a 2M sodium hydroxide coagulant solution (NaOH) and used in the form of wires and screens as a reinforcement structure to obtain the tubes. In order to characterize the tubes, optical microscopy, contact angle, degree of swelling, in vitro biodegradation, cytotoxicity and tensile strength were used. The results indicated that the tubes have uniformity over the entire length and as for the resistance to the trace, the tube reinforced with mesh presented greater deformation, while the tube reinforced with wire presented a higher value of rupture stress. The degree of swelling was higher in chitosan tubes with mesh. As for the biodegradation test, it was observed that the lysozyme samples showed greater loss of mass and the cytotoxicity test confirmed the cell viability of the material, concluding that the tubes reinforced with chitosan wires are promising for use in vascular surgeries.

Flow-driven Surface Instabilities of Tubular Chitosan Hydrogel.

Spatial structures break their symmetry under the influence of shear stress arising from fluid flow. Here, we present surface instabilities appearing on chitosan tubes when an acidic solution of chitosan with various molecular weight is injected into a pool of sodium hydroxide solution. At slow flow rates wrinkle‐to‐fold transition takes place along the direction of the flow yielding a banded structure. For greater injection rates we observe coexisting modes of wrinkles and folds which are stabilized to periodic wrinkles when the alkaline concentration is increased. The instabilities are characterized by the scaling laws of the pattern wavelength and amplitude with the tube characteristics. Our experimental adaptation of mechanical instabilities provides a new in situ method to create soft biomaterials with the desired surface morphology without the use of any prefabricated templates.

Self-assembling peptide hydrogels functionalized with LN- and BDNF- mimicking epitopes synergistically enhance peripheral nerve regeneration.

The regenerative capacity of the peripheral nervous system is closely related to the role that Schwann cells (SCs) play in construction of the basement membrane containing multiple extracellular matrix proteins and secretion of neurotrophic factors, including laminin (LN) and brain-derived neurotrophic factor (BDNF). Here, we developed a self-assembling peptide (SAP) nanofiber hydrogel based on self-assembling backbone Ac-(RADA)4-NH2 (RAD) dual-functionalized with laminin-derived motif IKVAV (IKV) and a BDNF-mimetic peptide epitope RGIDKRHWNSQ (RGI) for peripheral nerve regeneration, with the hydrogel providing a three-dimensional (3D) microenvironment for SCs and neurites.Methods: Circular dichroism (CD), atomic force microscopy (AFM), and scanning electron microscopy (SEM) were used to characterize the secondary structures, microscopic structures, and morphologies of self-assembling nanofiber hydrogels. Then the SC adhesion, myelination and neurotrophin secretion were evaluated on the hydrogels. Finally, the SAP hydrogels were injected into hollow chitosan tubes to bridge a 10-mm-long sciatic nerve defect in rats, and in vivo gene expression at 1 week, axonal regeneration, target muscular re-innervation, and functional recovery at 12 weeks were assessed.Results: The bioactive peptide motifs were covalently linked to the C-terminal of the self-assembling peptide and the functionalized peptides could form well-defined nanofibrous hydrogels capable of providing a 3D microenvironment similar to native extracellular matrix. SCs displayed improved cell adhesion on hydrogels with both IKV and RGI, accompanied by increased cell spreading and elongation relative to other groups. RSCs cultured on hydrogels with IKV and RGI showed enhanced gene expression of NGF, BDNF, CNTF, PMP22 and NRP2, and decreased gene expression of NCAM compared with those cultured on other three groups after a 7-day incubation. Additionally, the secretion of NGF, BDNF, and CNTF of RSCs was significantly improved on dual-functionalized peptide hydrogels after 3 days. At 1 week after implantation, the expressions of neurotrophin and myelin-related genes in the nerve grafts in SAP and Autograft groups were higher than that in Hollow group, and the expression of S100 in groups containing both IKV and RGI was significantly higher than that in groups containing either IKV or RGI hydrogels, suggesting enhanced SC proliferation. The morphometric parameters of the regenerated nerves, their electrophysiological performance, the innervated muscle weight and remodeling of muscle fibers, and motor function showed that RAD/IKV/RGI and RAD/IKV-GG-RGI hydrogels could markedly improve axonal regeneration with enhanced re-myelination and motor functional recovery through the synergetic effect of IKV and RGI functional motifs.Conclusions: We found that the dual-functionalized SAP hydrogels promoted RSC adhesion, myelination, and neurotrophin secretion in vitro and successfully bridged a 10-mm gap representing a sciatic nerve defect in rats in vivo. The results demonstrated the synergistic effect of IKVAV and RGI on axonal regrowth and function recovery after peripheral nerve injury.

Carbon microtubes derived from self-rolled chitosan acetate films and graphitized by joule heating

Herein, a fast and efficient approach is proposed to obtain carbon microtubes with graphitic structure. The precursor tubes are formed via self-rolling of chitosan acetate (CA) biopolymer films in water. The self-rolling capacity is imparted to the CA films by heating them at 150 °C on a hot plate for several minutes in the ambient atmosphere. The resulting differential swelling of the top and bottom layers of the films in water generates the bending moment. The amorphous carbon microtubes are produced in Ar atmosphere via thermal treatment of chitosan acetate microtubes at 900 °C. Carbon/oxygen ratio increases from 1.2 for the pristine chitosan tube to 5.3 after the pyrolysis process. Finite electrical conductivity of the so-prepared tubes (σ ≈ 1 · 103 S · m−1) allows their consecutive treatment by joule heating. The current-induced graphitization of the tubes at 2500 °C is confirmed by TEM, Raman spectroscopy and XRD techniques. Electrical conductivity of tubes raised in course of the treatment to (σ ≈ 1.8 · 104 S · m−1). In joule-heated tubes, the carbon/oxygen ratio increases to 9.4.

Chitosan tubes enriched with fresh skeletal muscle fibers for delayed repair of peripheral nerve defects

Nerve regeneration after delayed nerve repair is often unsuccessful. Indeed, the expression of genes associated with regeneration, including neurotrophic and gliotrophic factors, is drastically reduced in the distal stump of chronically transected nerves; moreover, Schwann cells undergo atrophy, losing their ability to sustain regeneration. In the present study, to provide a three-dimensional environment and trophic factors supporting Schwann cell activity and axon re-growth, we combined the use of an effective conduit (a chitosan tube) with a promising intraluminal structure (fresh longitudinal skeletal muscle fibers). This enriched conduit was used to repair a 10-mm rat median nerve gap after 3-month delay and functional and morphometrical analyses were performed 4 months after nerve reconstruction. Our data show that the enriched chitosan conduit is as effective as the hollow chitosan conduit in promoting nerve regeneration, and its efficacy is not statistically different from the autograft, considered the “gold standard” technique for nerve reconstruction. Since hollow tubes not always lead to good results after long defects (> 20 mm), we believe that the conduit enriched with fresh muscle fibers could be a promising strategy to repair longer gaps, as muscle fibers create a favorable three-dimensional environment and release trophic factors. All procedures were approved by the Bioethical Committee of the University of Torino and by the Italian Ministry of Health (approval number: 864/2016/PR) on September 14, 2016.

Effect of Local Administration of Verapamil Combined with Chitosan Based Hybrid Nanofiber Conduit on Transected Sciatic Nerve in Rat.

Objective:To assess the effect of locally administered verapamil on transected peripheral nerve regeneration and functional recovery.Methods:Sixty male healthy white Wistar rats were divided into four experimental groups (n=15), randomly: In transected group (TC), left sciatic nerve was transected and stumps were fixed in the adjacent muscle. In treatment group defect was bridged using chitosan tube (CHIT/Verapamil) filled with 10 µL verapamil (100ng/mL). In chitosan conduit group (CHIT), the tube was filled with phosphate-buffered saline alone. In sham-operated group (SHAM), sciatic nerve was exposed and manipulated. The repair trend was examined based on behavioral and performance tests as well as the variations of the gastrocnemius muscle, morphometric indices, and immunohistochemical indices.Results:Sciatic nerve functional study, muscle mass and morphometric indices confirmed faster recovery of regenerated axons in CHIT/Verapamil than CHIT group (P = 0.001). When loaded in a chitosan tube verapamil accelerated and improved functional recovery and morphometric indices of sciatic nerve. Immunohistochemical analysis revealed the S-100 protein was vastly present in the transverse nerve sections and the myelin sheath. In the treatment group (chit/verapamil), the immunohistochemical susceptibility of the axons being repaired and the axons in the myelin sheath to S-100 protein was higher than the other groups.Conclusion:The present study demonstrated that a single local application of verapamil could accelerate functional recovery after transection of sciatic nerve.

Améliorer l’issue des lésions nerveuses sensorielles traumatiques de la main par l’utilisation supplémentaire d’un tube nerveux de chitosane dans la réparation du nerf primaire — un essai bicentrique contrôlé randomisé

Peripheral sensory nerve injuries are a major problem of acute hand trauma surgery. Standard treatment is microsurgical end-to-end repair of these nerves whenever possible. If surgery fails, a permanent loss of sensibility and the formation of painful neuroma are consequences. The objective of this study was to evaluate whether the additional use of a chitosan nerve tube in primary nerve repair influences the sensory convalescence. A randomized controlled bicenter trial with parallel group design and double blinded assessment was conducted to show superiority of the additional use of a chitosan nerve tube compared to microsurgical nerve repair alone. 74 participants were enrolled. The ability of static 2-point-discrimanation (2PD) 6 months after surgery was the primary outcome parameter. Considerable secondary outcome parameters were filament recognition (Semmes Weinstein) testing, pain, appearance of neuroma and the Disabilities of the Arm, Shoulder and Hand (DASH)–score. In Intervention group both sensibility (expressed by 2PD) and tactile gnosis (expressed by Semmes Weinstein testing) could be increased significantly. Mean 2PD in control group at 6-month follow-up was 8 (2–20) mm and 6.3 (1–15) mm in intervention group (P = 0.029). 2PD correlated with DASH-score. In control group, 3 neuroma were found whereas none in intervention group. There were no implant-associated complications. Peripheral sensory nerve regeneration can be improved significantly by additional use of a chitosan nerve tube. An augmented ability of static 2-point discrimination has clinical relevancy.

Enhancing the Outcome of Traumatic Sensory Nerve Lesions of the Hand by Additional Use of a Chitosan Nerve Tube in Primary Nerve Repair: A Randomized Controlled Bicentric Trial.

Peripheral sensory nerve injuries present a significant yet common challenge in acute hand trauma surgery. Standard treatment remains microsurgical end-to-end nerve repair where appropriate. Permanent loss of sensitivity and painful neuroma formation are typical sequelae of unsuccessful surgery. The objective of this study was to evaluate whether the additional use of a chitosan nerve tube in primary nerve repair positively influences sensory recovery. A randomized, controlled, two-center trial with parallel group design and double-blind assessment was conducted to demonstrate the superiority of the additional use of a chitosan nerve tube compared with microsurgical nerve repair alone. Seventy-four participants were enrolled. The primary outcome parameter used was degree of static two-point discrimination at 6 months after surgery. Additional secondary outcome parameters included filament recognition testing (Semmes-Weinstein); pain; neuroma development; and the Disabilities of the Arm, Shoulder and Hand score. Nerve repair with additional use of chitosan nerve tubes (intervention group) significantly increased both tactile gnosis (expressed by two-point discrimination) and sensitivity (expressed by Semmes-Weinstein testing). The mean two-point discrimination at 6-month follow-up was 8 mm (range, 2 to 20 mm) in the control group and 6.3 mm (range, 1 to 15 mm) in the intervention group, respectively (p = 0.029). Two-point discrimination correlated with the Disabilities of the Arm, Shoulder and Hand score. In the control and intervention groups, respectively, three versus zero neuromas were found. Peripheral sensory nerve regeneration can be improved significantly by additional use of a chitosan nerve tube. An improved ability of static two-point discrimination is clinically relevant. Therapeutic, I.

Chitosan Tubes Enriched with Fresh Skeletal Muscle Fibers for Primary Nerve Repair.

Muscle-in-vein conduit is successfully employed for repairing nerve injuries: the vein prevents muscle fiber dispersion, while the muscle prevents the vein collapse and creates a favorable environment for Schwann cell migration and axon regrowth. However, it requires microsurgical skills. In this study we show a simple strategy to improve the performance of a chitosan hollow tube by the introduction of fresh skeletal muscle fibers. The hypothesis is to overcome the technical issue of the muscle-in-vein preparation and to take advantage of fiber muscle properties to create an easy and effective conduit for nerve regeneration. Rat median nerve gaps were repaired with chitosan tubes filled with skeletal muscle fibers (muscle-in-tube graft), hollow chitosan tubes, or autologous nerve grafts. Our results demonstrate that the fresh skeletal muscle inside the conduit is an endogenous source of soluble Neuregulin 1, a key factor for Schwann cell survival and dedifferentiation, absent in the hollow tube during the early phase of regeneration. However, nerve regeneration assessed at late time point was similar to that obtained with the hollow tube. To conclude, the muscle-in-tube graft is surgically easy to perform and we suggest that it might be a promising strategy to repair longer nerve gap or for secondary nerve repair, situations in which Schwann cell atrophy is a limiting factor for recovery.

A neurotrophic peptide-functionalized self-assembling peptide nanofiber hydrogel enhances rat sciatic nerve regeneration

Nerve guidance conduit (NGC) is a potential alternative to autologous nerve for peripheral nerve regeneration. A promising therapeutic strategy is to modify the nerve guidance conduit intraluminal microenvironment using physical and/or chemical guidance cues. In this study, a neurotrophic peptide-functionalized self-assembling peptide nanofiber hydrogel that could promote PC12 cell adhesion, proliferation, and neuronal differentiation in vitro was prefilled in the lumen of a hollow chitosan tube (hCST) to accelerate axonal regeneration in a rat sciatic nerve defect model. The functionalized self-assembling peptide was developed by introducing a neurotrophic peptide (RGI, RGIDKRHWNSQ) derived from brain-derived neurotrophic factor (BDNF) to the C-terminus of the self-assembling peptide RADA16-I (Ac-(RADA)4-CONH2). Morphological, histological, electrophysiological, and functional analyses demonstrated that the RGI-functionalized, self-assembling, peptide nanofiber hydrogel RAD/RGI could produce a neurotrophic microenvironment that markedly improved axonal regeneration with enhanced re-myelination and motor functional recovery.

Schwann cells and mesenchymal stem cells in laminin- or fibronectin-aligned matrices and regeneration across a critical size defect of 15 mm in the rat sciatic nerve.

OBJECTIVE Artificial nerve guides are being developed to substitute for autograft repair after peripheral nerve injuries. However, the use of conduits is limited by the length of the gap that needs to be bridged, with the success of regeneration highly compromised in long gaps. Addition of aligned proregenerative cells and extracellular matrix (ECM) components inside the conduit can be a good strategy to achieve artificial grafts that recreate the natural environment offered by a nerve graft. The purpose of this study was to functionalize chitosan devices with different cell types to support regeneration in limiting gaps in the rat peripheral nerve. METHODS The authors used chitosan devices combined with proteins of the ECM and cells in a rat model of sciatic nerve injury. Combinations of fibronectin and laminin with mesenchymal stem cells (MSCs) or Schwann cells (SCs) were aligned within tethered collagen-based gels, which were placed inside chitosan tubes that were then used to repair a critical-size gap of 15 mm in the rat sciatic nerve. Electrophysiology and algesimetry tests were performed to analyze functional recovery during the 4 months after injury and repair. Histological analysis was performed at the midlevel and distal level of the tubes to assess the number of regenerated myelinated fibers. RESULTS Functional analysis demonstrated that SC-aligned scaffolds resulted in 100% regeneration success in a 15-mm nerve defect in this rat model. In contrast, animals that underwent repair with MSC-aligned constructs had only 90% regeneration success, and those implanted with acellular bridges had only 75% regeneration success. CONCLUSIONS These results indicate that the combination of chitosan conduits with ECM-enriched cellular gels represents a good alternative to the use of autografts for repairing long nerve gaps.

Chitosan-based hydrogels for developing a small-diameter vascular graft: in vitro and in vivo evaluation

Aims. Vascular grafts made of synthetic polymers perform poorly in small-diameter applications (cardiac and peripheral bypass). Chitosan is a biocompatible natural polymer that can provide a novel biological scaffold for tissue engineering development. The goal of this study was to demonstrate the biocompatibility of a novel chitosan preparation in vitro and in vivo, and to assess its potential as a scaffold for vascular applications. Methods and results. A series of experiments of increasing complexity, ranging from in vitro biocompatibility and hemocompatibility tests to in vivo studies in small and large animals (rats and sheep), was performed to provide a comprehensive analysis of chitosan hydrogels’ biological properties. In vitro studies established that: (i) chitosan supported human endothelial progenitor cells adhesion, proliferation and resistance to physiological shear stress; (ii) chitosan did not activate platelets, the complement system, or the intrinsic coagulation pathway. In vivo results showed: (iii) no resorption of chitosan and no chronic inflammation at 60 days in a rat heterotopic implantation model (magnetic resonance imaging and histology); (iv) no flow obstruction (Doppler ultrasound) and no thrombus formation (histology and scanning electron microscopy) at 2 h after a carotid arteriotomy repair with chitosan patches in sheep. Finally, two chitosan tubes were implanted as carotid interposition grafts for 3 days in sheep showing that chitosan was strong enough to be sutured, to withstand arterial pressure, and no flow obstruction was observed through this short period. Conclusion. Chitosan-based hydrogels displayed promising in vitro biocompatibility and hemocompatibility properties as well as in vivo short-term performance.