Repair Of Injured Peripheral Nerves Research Articles

An Adaptable Drug Delivery System Facilitates Peripheral Nerve Repair by Remodeling the Microenvironment.

The multifaceted process of nerve regeneration following damage remains a significant clinical issue, due to the lack of a favorable regenerative microenvironment and insufficient endogenous biochemical signaling. However, the current nerve grafts have limitations in functionality, as they require a greater capacity to effectively regulate the intricate microenvironment associated with nerve regeneration. In this regard, we proposed the construction of a functional artificial scaffold based on a "two-pronged" approach. The whole system was developed by encapsulating Tazarotene within nanomicelles formed through self-assembly of reactive oxygen species (ROS)-responsive amphiphilic triblock copolymer, all of which were further loaded into a thermosensitive injectable hydrogel. Notably, the hydrogel exhibits obvious temperature sensitivity at a concentration of 6 wt %, and the nanoparticles possess concentration-dependent H2O2-response capability with a controlled release profile in 48 h. The combined strategy promoted the repair of injured peripheral nerves, attributed to the dual role of the materials, which mainly involved providing structural support, modulating the immune microenvironment, and enhancing angiogenesis. Overall, this study opens up intriguing prospects in tissue engineering.

Brief Electrical Stimulation Promotes Recovery after Surgical Repair of Injured Peripheral Nerves.

Injured peripheral nerves regenerate their axons in contrast to those in the central nervous system. Yet, functional recovery after surgical repair is often disappointing. The basis for poor recovery is progressive deterioration with time and distance of the growth capacity of the neurons that lose their contact with targets (chronic axotomy) and the growth support of the chronically denervated Schwann cells (SC) in the distal nerve stumps. Nonetheless, chronically denervated atrophic muscle retains the capacity for reinnervation. Declining electrical activity of motoneurons accompanies the progressive fall in axotomized neuronal and denervated SC expression of regeneration-associated-genes and declining regenerative success. Reduced motoneuronal activity is due to the withdrawal of synaptic contacts from the soma. Exogenous neurotrophic factors that promote nerve regeneration can replace the endogenous factors whose expression declines with time. But the profuse axonal outgrowth they provoke and the difficulties in their delivery hinder their efficacy. Brief (1 h) low-frequency (20 Hz) electrical stimulation (ES) proximal to the injury site promotes the expression of endogenous growth factors and, in turn, dramatically accelerates axon outgrowth and target reinnervation. The latter ES effect has been demonstrated in both rats and humans. A conditioning ES of intact nerve days prior to nerve injury increases axonal outgrowth and regeneration rate. Thereby, this form of ES is amenable for nerve transfer surgeries and end-to-side neurorrhaphies. However, additional surgery for applying the required electrodes may be a hurdle. ES is applicable in all surgeries with excellent outcomes.

Electroconductive and Immunomodulatory Natural Polymer‐Based Hydrogel Bandages Designed for Peripheral Nerve Regeneration

AbstractSuccessful regeneration of the peripheral nerve relies on the collaborative efforts of neural cells and immune cells. Conductive hydrogels have yielded promising results in supporting axonal growth; however, their inability to regulate the immune response and their poor biological integration with tissues hinder the repair of injured peripheral nerves. Herein, an adhesive conductive immunomodulatory nerve hydrogel bandage is developed for nerve regeneration. The nerve bandage hydrogel is prepared from the bioactive material extracellular matrix (ECM), oxidized polysaccharides, and poly(3,4‐ethylenedioxythiophene)/poly(styrenesulfonate) (PEDOT:PSS) by self‐assembly and dynamic Schiff base cross‐linking. The drug indole‐3‐propionic acid (IPA) is loaded into nerve bandages and contributes to the rapid chemotaxis of neutrophils in the dorsal root ganglia and modulation of the immune system. In addition, the conductive hydrogel bandage exhibits close conformal contact with the injured nerve, forming a stable and tightly coupled electrical bridge with the electroresponsive neural tissue. In summary, the nerve bandage effectively promote nerve regeneration and enable both anatomical and functional recovery of neural tissue while preventing muscle atrophy. This work provides a new strategy for peripheral nerve regeneration and may have critical clinical applications in the future.

Engineering a wirelessly self-powered and electroconductive scaffold to promote peripheral nerve regeneration

Electroactive biomaterials have been shown useful for the repair of injured peripheral nerve. While for conventional conductive conduits, outer electrical stimulation device is unavoidably employed to exert electrical signals, and their rigid microstructures are usually incompatible with neural cells. Herein, a soft carbon nanotubes@gelatin methacryloyl/poly(L-lactic acid) (CNTs@GelMA/PLLA) nerve tissue-engineering scaffold was fabricated, which provided an endogenous piezoelectric stimulation and conductive microenvironment. Based on amounts of in-vitro experiment data, such composite scaffold significantly improved adhesion and elongation of Schwann cells, and meanwhile promoted axonal outgrowth and neurites number of dorsal root ganglions. More interestingly, the scaffold was applied to a 10-mm sciatic nerve defect in rats and harvested at 12 weeks post-implantation. Immunohistochemical staining results indicated that our proposed graft significantly facilitated peripheral nerve regeneration by promoting myelination and axon outgrowth, meanwhile an enhanced motor functional recovery caused by the scaffold was also revealed due to the obviously-improved sciatic functional index and muscle weights. Overall, the soft, self-powered, and electroconductive CNTs@GelMA/PLLA scaffold is a promising candidate for the treatment of peripheral nerve injuries.

Bone Marrow Derived Cells Mediated Regulation of Schwann Cells Proliferation and Migration In vitro

Schwann cells (SCs) are the unique glial cells in the peripheral nerve. After peripheral nerve injury, SCs change their morphology, function, and play a new role as repair cells, through this process, factors that enhance SCs proliferation and promote SCs migration will benefit nerve regeneration and functional recovery. Bone marrow-derived cells (BMDCs), which has been proved to offer several regenerative benefits for tissue and organ injuries. However, whether BMDCs support nerve regeneration by affect the proliferation and migration of SCs, remains unclear. Here, we taken BMDCs co-cultured with nerve segments, and then investigated the proliferation and migration of SCs In vitro. Results demonstrated that BMDCs can enhance the proliferation of SCs, and then promoted SCs migration, forming Bungner’s brand-liked structure In vitro. These finding provide a potential new method for repair of injured peripheral nerves in clinical.

Effects of 17-β-estradiol released from shape-memory terpolymer rods on sciatic nerve regeneration after injury and repair with chitosan nerve conduit in female rats.

The aim of this study was to assess 17-β-estradiol (E2) influence on sciatic nerve regeneration after injury followed by a repair with chitosan conduit in ovariectomized female rats. The study was performed in 2 groups (n = 16) of rats: OVChit - after excision of a fragment of the sciatic nerve, a chitosan conduit was implanted; OVChitE10 group - additionally to chitosan conduit, shape-memory terpolymer rods based on poly(L-lactide-co-glycolide- co-trimethylene carbonate) releasing 17-β-estradiol for 20 weeks were implanted. The mean number of regenerating axons and mean fiber area were significantly greater in 17-β-estradiol-treated animals. In this group, the infiltrate of leukocytes was diminished. The presence of 17-β-estradiol receptors alpha and beta in motoneurons in the spinal cord were discovered. This may indicate the location where 17-β-estradiol affects the regeneration of the injured nerve. Estradiol released from the terpolymer rods for 20 weeks could enhance, to some extent, sciatic nerve regeneration after injury, and diminish the inflammatory reaction. In the future, 17-β-estradiol entrapped in terpolymer rods could be used in the repair of injured peripheral nerves, but there is a need for further studies.

Low-Stiffness Hydrogels Promote Peripheral Nerve Regeneration Through the Rapid Release of Exosomes.

A hydrogel system loaded with mesenchymal stem cell–derived exosome (MSC-Exos) is an attractive new tool for tissue regeneration. However, the effect of the stiffness of exosome-loaded hydrogels on tissue regeneration is unclear. Here, the role of exosome-loaded hydrogel stiffness, during the regeneration of injured nerves, was assessed in vivo. The results showed that the photocrosslinkable hyaluronic acid methacrylate hydrogel stiffness plays an important role in repairing nerve injury. Compared with the stiff hydrogels loaded with exosomes, soft hydrogels loaded with exosomes showed better repair of injured peripheral nerves. The soft hydrogel promoted nerve repair by quickly releasing exosomes to inhibit the infiltration of macrophages and the expression of the proinflammatory factors IL-1β and TNF-α in injured nerves. Our work revealed that exosome-loaded hydrogel stiffness plays an important role in tissue regeneration by regulating exosome release behavior and provided important clues for the clinical application of biological scaffold materials.

Based on proteomics to explore the mechanism of mecobalamin promoting the repair of injured peripheral nerves.

Mecobalamin is commonly used in the adjuvant intervention of various peripheral nerve injuries. Actin cytoskeleton plays a role in the regeneration of myelin and axon. Therefore, the purpose of this study was to explore the possibility of mecobalamin regulating actin cytoskeleton in repairing nerve injury. In this study, a crush injury on the right sciatic nerve of two groups of rats (12 in each group) was established. The control group was only given normal saline (i.g.), and the intervention group was given mecobalamin 1mg/kg(i.g.).Theratswere sacrificed on 28th day and the injured nerves were collected for proteomics. The result shows that regulation of actin cytoskeleton pathway changed significantly. The expression of protein Vav1 was verified by Western blot and immunofluorescence. In the intervention group, the nerve fiber structure was complete, the axons were dense and symmetrical, and the myelin sheath was compact and uniform in thickness. The positive rate of myelin basic protein and βⅢ-tubulin was higher than that in the control group. The findings of the study show that mecobalamin regulates the actin cytoskeleton in the repair of nerve damage and upregulates Vav1 in the regulation of actin cytoskeleton pathway.

Research Progress in the Repair of Peripheral Nerve Injury with Adipose-Derived Stem Cell Exosomes

The repair of peripheral nerve injury has always been a difficult clinical problem. Although a variety of treatment methods are available in clinical practice, their efficacy is limited. In recent years, the components carried by adipose stem cell exosomes and their functions have been increasingly discovered. A large number of experiments conducted around the world have shown that adipose-derived stem cell exosomes have a positive effect on the repair of peripheral nerve injury. This article reviews recent progress toward the use of adipose-derived stem cell exosomes in the repair of injured peripheral nerves and possible future research directions involving adipose-derived stem cell exosomes.

Neutrophil peptide-1 promotes the repair of sciatic nerve injury through the expression of proteins related to nerve regeneration

ABSTRACT Objectives Small-molecule polypeptide neutrophil peptide 1 (NP-1) was reported to promote the regeneration of the sciatic nerve after denervation, but the mechanisms underlying this effect of NP-1 are unclear. Here, we established a Sprague-Dawley rat model of crush injury to study the effect of a single intermuscular injection of NP-1 on the repair of injured peripheral nerves and elucidate the possible underlying mechanism. Methods 39 rats were randomly selected to join this study and divided into the blank control group (normal group, n=9), experimental group (NP-1 group, n=15), and negative control group (NS group, n=15). The dynamic expression of cytokines in different groups of nerve tissues during Wallerian degeneration was observed using protein chips at different time points after injury. Recovery of injured nerves was determined based on the general condition, local gross morphology of the nerve suture site, sciatic nerve function index, neuroelectrophysiology, and osmic acid staining at 6 weeks after the surgery. The recovery of effector function was determined based on wet weight, hematoxylin-eosin staining, modified Gomori staining, and nicotinamide adenine dinucleotide-tetrazolium reductase staining at 6 weeks after the surgery. Results It was found that a single topical administration of NP-1 promoted sciatic nerve regeneration after crush injury and affected the expression of proteins related to neurotrophy, inflammation, cell chemotaxis, and cell generation pathways.

Therapeutic strategies for peripheral nerve injury: decellularized nerve conduits and Schwann cell transplantation.

In recent years, the use of Schwann cell transplantation to repair peripheral nerve injury has attracted much attention. Animal-based studies show that the transplantation of Schwann cells in combination with nerve scaffolds promotes the repair of injured peripheral nerves. Autologous Schwann cell transplantation in humans has been reported recently. This article reviews current methods for removing the extracellular matrix and analyzes its composition and function. The development and secretory products of Schwann cells are also reviewed. The methods for the repair of peripheral nerve injuries that use myelin and Schwann cell transplantation are assessed. This survey of the literature data shows that using a decellularized nerve conduit combined with Schwann cells represents an effective strategy for the treatment of peripheral nerve injury. This analysis provides a comprehensive basis on which to make clinical decisions for the repair of peripheral nerve injury.

Nerve wrap after end-to-end and tension-free neurorrhaphy attenuates neuropathic pain: A prospective study based on cohorts of digit replantation

The repair of injured peripheral nerve is still challenging for surgeons. The end-to-end and tension-free neurorrhaphy is the current gold standard for reconstruction after complete nerve transection without significant defect. The main objective of this study neurorrhaphy in digit replantation affects the sensory recovery and neuropathic pain in replanted digit. Total 101 patients who received replantation of single completely amputated digit were included for analysis in this study. In group I (n = 49), the digital nerves were repaired with end-to-end and tension-free neurorrhaphy and then wrapped into a tendon-derived collagen nerve conduit. In group II (n = 52), the digital nerves were repaired with end-to-end and tension-free neurorrhaphy only. The static two-point discrimination (s2PD) was performed to evaluate sensory recovery. Visual analog scale (VAS) scores of pain at rest and with exertion were measured respectively. The s2PD tests at three and six months after surgery did not show any significant difference between the two groups. The VAS scores at rest and with exertion of group I were significantly reduced compared with those of group II at three and six months after surgery. Thus, we concluded that nerve wrap into a collagen conduit after end-to-end and tension-free neurorrhaphy could attenuate neuropathic pain after digit replantation but have no benefit for sensory recovery.

Muscle-Derived Stem Cells in Peripheral Nerve Regeneration: Reality or Illusion?

Owing to the complicated and time-consuming regenerative process, the repair of injured peripheral nerves depends largely on ongoing stem-cell therapy. Decades ago, researchers successfully isolated and identified muscle-derived stem cells (MDSCs) and discovered their potential for multidifferentiation. MDSCs play an important role in trauma repair associated with neuromuscular and vascular injury by simultaneously promoting tissue regrowth via direct differentiation and systematic secretion under physiological conditions. However, the isolation, culture, induction and application of MDSCs require further methodological analysis before clinical application. In this review, we comprehensively discuss the challenges associated with neural regeneration and reviewed the progress of stem cell based regenerative medicine, in an effort to realize the potential of MDSCs in nerve regeneration.

Recent Approaches for Augmenting Peripheral Nerve Regeneration: Mini Review

Injury to superficial or deeply seated nerve is commonly reported in human and animals causing crippling morbidity. The sciatic nerve is the most frequently involved nerve in such injuries. There could be several approaches for the repair of injured peripheral nerves. However, recent therapeutic approaches include the use of molecular, cellular and electrophysical methods.Mesenchymal stem cells are the most commonly used adult stem cells for the therapeutic purpose. Several studies have shown that stem cell transplantation may promote neural regeneration by enhanced growth factor secretion, extracellular matrix production and differentiation into Schwann cell, which are primarily responsible for the axonal regeneration. Scaffolds are used to maintain cell viability, support cell proliferation and permit intercellular communication. ECM proteins, and nerve growth factors can be incorporated into nerve conduits in order to improve the nerve regenerative ability. Among the electro physical methods use of 20Hz electrical stimulation, No thermal Laser Amnion Wrap and Thermal Laser Welding have shown promising results. The present review focuses on the application and outcome of important molecular, cellular and electro physical methods used for nerve regeneration.

Brain injury in combination with tacrolimus promotes the regeneration of injured peripheral nerves.

Both brain injury and tacrolimus have been reported to promote the regeneration of injured peripheral nerves. In this study, before transection of rat sciatic nerve, moderate brain contusion was (or was not) induced. After sciatic nerve injury, tacrolimus, an immunosuppressant, was (or was not) intraperitoneally administered. At 4, 8 and 12 weeks after surgery, Masson's trichrome, hematoxylin-eosin, and toluidine blue staining results revealed that brain injury or tacrolimus alone or their combination alleviated gastrocnemius muscle atrophy and sciatic nerve fiber impairment on the experimental side, simultaneously improved sciatic nerve function, and increased gastrocnemius muscle wet weight on the experimental side. At 8 and 12 weeks after surgery, brain injury induction and/or tacrolimus treatment increased action potential amplitude in the sciatic nerve trunk. Horseradish peroxidase retrograde tracing revealed that the number of horseradish peroxidase-positive neurons in the anterior horn of the spinal cord was greatly increased. Brain injury in combination with tacrolimus exhibited better effects on repair of injured peripheral nerves than brain injury or tacrolimus alone. This result suggests that brain injury in combination with tacrolimus promotes repair of peripheral nerve injury.

NECL1 coated PLGA as favorable conduits for repair of injured peripheral nerve

Restoration of normal neurological function of transected peripheral nerve challenged regenerative medicine and surgery. Previous studies showed that Nectin-like molecule 1 (NECL1) is one of the important adhesion molecules on the axons and Schwann cells is located along the internodes in direct apposition to NECL1. In this study, we fabricated PLGA membrane pre-coated with NECL1, mimicking the natural axons to enhance the adhesion of Schwann cells. Investigation of the cellular response in vitro was performed by detecting cytotoxicity, proliferation, morphology, viability, specific markers and Scanning Electron Microscopy (SEM) of Schwann cells cultured in PLGA. Further, the NECL1-coated PLGA conduits were used for peripheral nerve repair after sciatic nerve defect was constructed. Results showed that PLGA-coated NECL1 enhanced cell proliferation compared with PLGA, as evidenced by MTT analysis, cell viability assay and histological evaluation. RT-PCR results showed that GDNF (glial cell line-derived neurotrophic factor), BDNF (brain-derived neurotrophic factor), CNTF (ciliary neurotrophic factor) and neurotrophic factors of axonal regeneration were highly expressed in PLGA/NECL1 group. S100, which is Schwann cell marker, was also elevated in PLGA-NCEL1 in both mRNA and protein expression as demonstrated by PCR and immunohistochemical examination. Moreover, in vivo study showed that implantation of PLGA/NCEL1 tubes in bridging the nerve defect can significantly improve Schwann cell aggregation and attachment and greatly enhance the functional recovery of nerve regeneration as compared with control and PLGA groups. Therefore, the novel blend of PLGA/NECL1 conduits proved to be promising candidate for tissue engineering scaffold.

Magnetic resonance imaging monitoring dual-labeled stem cells for treatment of mouse nerve injury

Background aimsAdipose-derived stem cells (ADSCs) have shown great promise in the regenerative repair of injured peripheral nerves. Magnetic resonance imaging (MRI) has provided attractive advantages in tracking superparamagnetic iron oxide nanoparticle (SPION)-labeled cells and evaluating their fate after cell transplantation. This study investigated the feasibility of the use of MRI to noninvasively track ADSCs repair of peripheral nerve injury in vivo. MethodsGreen fluorescent protein (GFP)-expressing ADSCs were isolated, expanded, differentiated into an SC-like phenotype (GFP-dADSCs) at early passages and subsequently labeled with SPIONs. The morphological and functional properties of the GFP-dADSCs were assessed through the use of immunohistochemistry. The intracellular stability, proliferation and viability of the labeled cells were evaluated in vitro. Through the use of a microsurgical procedure, the labeled cells were then seeded into sciatic nerve conduits in C57/BL6 mice to repair a 1-cm sciatic nerve gap. A clinical 3-T MRI was performed to investigate the GFP-dADSCs in vitro and the transplanted GFP-dADSCs inside the sciatic nerve conduits in vivo. ResultsThe GFP-dADSCs were efficiently labeled with SPIONs, without affecting their viability and proliferation. The labeled cells implanted into the mice sciatic nerve conduit exhibited a significant increase in axonal regeneration compared with the empty conduit and could be detected by MRI. Fluorescent microscopic examination, histological analysis and immunohistochemistry confirmed the axon regeneration and MRI results. ConclusionsThese data will elucidate the neuroplasticity of ADSCs and provide a new protocol for in vivo tracking of stem cells that are seeded to repair injured peripheral nerves.

Engineering a multimodal nerve conduit for repair of injured peripheral nerve

Injury to nerve tissue in the peripheral nervous system (PNS) results in long-term impairment of limb function, dysaesthesia and pain, often with associated psychological effects. Whilst minor injuries can be left to regenerate without intervention and short gaps up to 2 cm can be sutured, larger or more severe injuries commonly require autogenous nerve grafts harvested from elsewhere in the body (usually sensory nerves). Functional recovery is often suboptimal and associated with loss of sensation from the tissue innervated by the harvested nerve. The challenges that persist with nerve repair have resulted in development of nerve guides or conduits from non-neural biological tissues and various polymers to improve the prognosis for the repair of damaged nerves in the PNS. This study describes the design and fabrication of a multimodal controlled pore size nerve regeneration conduit using polylactic acid (PLA) and (PLA):poly(lactic-co-glycolic) acid (PLGA) fibers within a neurotrophin-enriched alginate hydrogel. The nerve repair conduit design consists of two types of PLGA fibers selected specifically for promotion of axonal outgrowth and Schwann cell growth (75:25 for axons; 85:15 for Schwann cells). These aligned fibers are contained within the lumen of a knitted PLA sheath coated with electrospun PLA nanofibers to control pore size. The PLGA guidance fibers within the nerve repair conduit lumen are supported within an alginate hydrogel impregnated with neurotrophic factors (NT-3 or BDNF with LIF, SMDF and MGF-1) to provide neuroprotection, stimulation of axonal growth and Schwann cell migration. The conduit was used to promote repair of transected sciatic nerve in rats over a period of 4 weeks. Over this period, it was observed that over-grooming and self-mutilation (autotomy) of the limb implanted with the conduit was significantly reduced in rats implanted with the full-configuration conduit compared to rats implanted with conduits containing only an alginate hydrogel. This indicates return of some feeling to the limb via the fully-configured conduit. Immunohistochemical analysis of the implanted conduits removed from the rats after the four-week implantation period confirmed the presence of myelinated axons within the conduit and distal to the site of implantation, further supporting that the conduit promoted nerve repair over this period of time. This study describes the design considerations and fabrication of a novel multicomponent, multimodal bio-engineered synthetic conduit for peripheral nerve repair.

The therapeutic potential of ex vivo expanded CD133+ cells derived from human peripheral blood for peripheral nerve injuries

CD133(+) cells have the potential to enhance histological and functional recovery from peripheral nerve injury. However, the number of CD133(+) cells safely obtained from human peripheral blood is extremely limited. To address this issue, the authors expanded CD133(+) cells derived from human peripheral blood using the serum-free expansion culture method and transplanted these ex vivo expanded cells into a model of sciatic nerve defect in rats. The purpose of this study was to determine the potential of ex vivo expanded CD133(+) cells to induce or enhance the repair of injured peripheral nerves. Phosphate-buffered saline (PBS group [Group 1]), 10(5) fresh CD133(+) cells (fresh group [Group 2]), 10(5) ex vivo expanded CD133(+) cells (expansion group [Group 3]), or 10(4) fresh CD133(+) cells (low-dose group [Group 4]) embedded in atelocollagen gel were transplanted into a silicone tube that was then used to bridge a 15-mm defect in the sciatic nerve of athymic rats (10 animals per group). At 8 weeks postsurgery, histological and functional evaluations of the regenerated tissues were performed. After 1 week of expansion culture, the number of cells increased 9.6 ± 3.3-fold. Based on the fluorescence-activated cell sorting analysis, it was demonstrated that the initial freshly isolated CD133(+) cell population contained 93.22% ± 0.30% CD133(+) cells and further confirmed that the expanded cells had a purity of 59.02% ± 1.58% CD133(+) cells. However, the histologically and functionally regenerated nerves bridging the defects were recognized in all rats in Groups 2 and 3 and in 6 of 10 rats in Group 4. The nerves did not regenerate to bridge the defect in any of the rats in Group 1. The authors' results show that ex vivo expanded CD133(+) cells derived from human peripheral blood have a therapeutic potential similar to fresh CD133(+) cells for peripheral nerve injuries. The ex vivo procedure that can be used to expand CD133(+) cells without reducing their function represents a novel method for developing cell therapy for nerve defects in a clinical setting.

A multipotent clonal human periodontal ligament cell line with neural crest cell phenotypes promotes neurocytic differentiation, migration, and survival

Repair of injured peripheral nerve is thought to play important roles in tissue homeostasis and regeneration. Recent experiments have demonstrated enhanced functional recovery of damaged neurons by some types of somatic stem cells. It remains unclear, however, if periodontal ligament (PDL) stem cells possess such functions. We recently developed a multipotent clonal human PDL cell line, termed cell line 1-17. Here, we investigated the effects of this cell line on neurocytic differentiation, migration, and survival. This cell line expressed the neural crest cell marker genes Slug, SOX10, Nestin, p75NTR, and CD49d and mesenchymal stem cell-related markers CD13, CD29, CD44, CD71, CD90, CD105, and CD166. Rat adrenal pheochromocytoma cells (PC12 cells) underwent neurocytic differentiation when co-cultured with cell line 1-17 or in conditioned medium from cell line 1-17 (1-17CM). ELISA analysis revealed that 1-17CM contained approximately 50 pg/ml nerve growth factor (NGF). Cell line 1-17-induced migration of PC12 cells, which was inhibited by a neutralizing antibody against NGF. Furthermore, 1-17CM exerted antiapoptotic effects on differentiated PC12 cells as evidenced by inhibition of neurite retraction, reduction in annexin V and caspase-3/7 staining, and induction of Bcl-2 and Bcl-xL mRNA expression. Thus, cell line 1-17 promoted neurocytic differentiation, migration, and survival through secretion of NGF and possibly synergistic factors. PDL stem cells may play a role in peripheral nerve reinnervation during PDL regeneration.