Biomaterials and tissue engineering in traumatic brain injury:novel perspectives on promoting neural regeneration

Traumatic brain injury is a serious medical condition that can be attributed to falls, motor vehicle accidents, sports injuries and acts of violence, causing a series of neural injuries and neuropsychiatric symptoms. However, limited accessibility to the injury sites, complicated histological and anatomical structure, intricate cellular and extracellular milieu, lack of regenerative capacity in the native cells, vast variety of damage routes, and the insufficient time available for treatment have restricted the widespread application of several therapeutic methods in cases of central nervous system injury. Tissue engineering and regenerative medicine have emerged as innovative approaches in the field of nerve regeneration. By combining biomaterials, stem cells, and growth factors, these approaches have provided a platform for developing effective treatments for neural injuries, which can offer the potential to restore neural function, improve patient outcomes, and reduce the need for drugs and invasive surgical procedures. Biomaterials have shown advantages in promoting neural development, inhibiting glial scar formation, and providing a suitable biomimetic neural microenvironment, which makes their application promising in the field of neural regeneration. For instance, bioactive scaffolds loaded with stem cells can provide a biocompatible and biodegradable milieu. Furthermore, stem cells-derived exosomes combine the advantages of stem cells, avoid the risk of immune rejection, cooperate with biomaterials to enhance their biological functions, and exert stable functions, thereby inducing angiogenesis and neural regeneration in patients with traumatic brain injury and promoting the recovery of brain function. Unfortunately, biomaterials have shown positive effects in the laboratory, but when similar materials are used in clinical studies of human central nervous system regeneration, their efficacy is unsatisfactory. Here, we review the characteristics and properties of various bioactive materials, followed by the introduction of applications based on biochemistry and cell molecules, and discuss the emerging role of biomaterials in promoting neural regeneration. Further, we summarize the adaptive biomaterials infused with exosomes produced from stem cells and stem cells themselves for the treatment of traumatic brain injury. Finally, we present the main limitations of biomaterials for the treatment of traumatic brain injury and offer insights into their future potential.

Low-temperature 3D-printed collagen/chitosan scaffolds loaded with exosomes derived from neural stem cells pretreated with insulin growth factor-1 enhance neural regeneration after traumatic brain injury

There are various clinical treatments for traumatic brain injury,including surgery,drug therapy,and rehabilitation therapy;howeve r,the therapeutic effects are limited.Scaffolds combined with exosomes represent a promising but challenging method for improving the repair of traumatic brain injury.In this study,we determined the ability of a novel 3D-printed collagen/chitosan scaffold loaded with exosomes derived from neural stem cells pretreated with insulin-like growth factor-1(3D-CC-INEXOS) to improve traumatic brain injury repair and functional recove ry after traumatic brain injury in rats.Composite scaffolds comprising collagen,chitosan,and exosomes derived from neural stem cells pretreated with insulin-like growth fa ctor-1(INEXOS) continuously released exosomes for 2weeks.Transplantation of 3D-CC-INExos scaffolds significantly improved motor and cognitive functions in a rat traumatic brain injury model,as assessed by the Morris water maze test and modified neurological seve rity scores.In addition,immunofluorescence staining and transmission electron microscopy showed that3D-CC-INExos implantation significantly improved the recove ry of damaged nerve tissue in the injured area.In conclusion,this study suggests that transplanted3D-CC-INExos scaffolds might provide a potential strategy for the treatment of traumatic brain injury and lay a solid foundation for clinical translation.

3D printing of functional bioengineered constructs for neural regeneration: a review

Three-dimensional(3D)printing technology has opened a new paradigm to controllably and reproducibly fabricate bioengineered neural constructs for potential applications in repairing injured nervous tissues or producing in vitro nervous tissue models.However,the complexity of nervous tissues poses great challenges to 3D-printed bioengineered analogues,which should possess diverse architectural/chemical/electrical functionalities to resemble the native growth microenvironments for functional neural regeneration.In this work,we provide a state-of-the-art review of the latest development of 3D printing for bioengineered neural constructs.Various 3D printing techniques for neural tissue-engineered scaffolds or living cell-laden constructs are summarized and compared in terms of their unique advantages.We highlight the advanced strategies by integrating topographical,biochemical and electroactive cues inside 3D-printed neural constructs to replicate in vivo-like microenvironment for functional neural regeneration.The typical applications of 3D-printed bioengineered constructs for in vivo repair of injured nervous tissues,bio-electronics interfacing with native nervous system,neural-on-chips as well as brain-like tissue models are demonstrated.The challenges and future outlook associated with 3D printing for functional neural constructs in various categories are discussed.

Molecular actions guiding neural regeneration in planarian

Planarian is among the simplest animals that possess a centralized nervous system （CNS）, and its neural regenera- tion involves the replacement of cells lost to normal ＇wear and tear＇ （cell turnover）, and/or injury. In this review, we state and discuss the recent studies on molecular control of neural regeneration in planarians. The spatial and temporal expression patterns of genes in intact and regenerating planarian CNS have already been described relatively clearly. The bone morphogenetic protein （BMP） and Wnt signaling pathways are identified to regulate neural regeneration. During neural regeneration, conserved axon guidance mechanisms are necessary for proper wiring of the nervous system. In addition, apoptosis may play an important role in controlling cell numbers, eliminating unnecessary tissues or cells and remodeling the old tissues for regenerating CNS. The bilateral symmetry is established by determination of anterior-posterior （A-P） and dorsal-ventral （D-V） patterns. Moreover, neurons positive to dopamine, serotonin （5-HT）, and gamma-aminobutyric acid （GABA） have been detected in planarians. Therefore, planarians present us with new, experimentally accessible contexts to study the molecular actions guiding neural regeneration.

Rho/ROCK pathway and neural regeneration: a potential therapeutic target for central nervous system and optic nerve damage

Rho-associated kinase (ROCK) is a serine/threonine kinase and one of the major downstream effectors of the small GTPase RhoA. The Rho/ROCK pathway is closely related to the pathogenesis of several central nervous system (CNS) disorders, and involved in many aspects of neuronal functions including neurite outgrowth and retraction. In the adult CNS, the damaged neuron regeneration is very difficult due to the presence of myelin-associated axon growth inhibitors such as Nogo, myelin-associated glycoprotein (MAG) and oligodendrocyte-myelin glycoprotein (Omgp), etc. The effects of these axon growth inhibitors are reversed by blocking the Rho/ROCK pathway 47 vitro, and the inhibition of Rho/ROCK pathway can promote axon regeneration and functional recovery in the injured CNS in viva In addition, the therapeutic effects of the Rho/ROCK inhibitors have also been demonstrated in some animal models and the Rho/ROCK pathway becomes an attractive target for the development of drugs for treating CNS disorders. In this review, we summarized on the effect of the Rho and the downstream factor ROCK in neural regeneration, and the potential therapeutic effect of Rho/ROCK inhibitors in the survival and axonal regeneration of retinal ganglion cell was also discussed.

Intravenous transplantation of bone marrow mesenchymal stem cells promotes neural regeneration after traumatic brain injury

To investigate the supplement of lost nerve cells in rats with traumatic brain injury by intravenous administration of allogenic bone marrow mesenchymal stem cells, this study established a Wistar rat model of traumatic brain injury by weight drop impact acceleration method and administered 3 × 106 rat bone marrow mesenchymal stem cells via the lateral tail vein. At 14 days after cell transplantation, bone marrow mesenchymal stem cells differentiated into neurons and astrocytes in injured rat cerebral cortex and rat neurological function was improved significantly. These findings suggest that intravenously administered bone marrow mesenchymal stem cells can promote nerve cell regeneration in injured cerebral cortex, which supplement the lost nerve cells.

Scientific progress regarding neural regeneration in the Web of Science A 10-year bibliometric analysis

BACKGROUND： Neural regeneration following nerve injury is an emerging field that attracts ex- tending interests all over the world. OBJECTIVE： To use bibliometric indexes to track studies focusing on neural regeneration, and to investigate the relationships among geographic origin, countries and institutes, keywords in the published articles, and especially focus on the region distribution, institution distribution, as well as collaborations in Chinese papers indexed in the Web of Science. METHODS： A list of neural regeneration studies was generated by searching the database of the Web of Science-Expanded using the term ＂Neural Regenera＊＂. Inclusive criteria： （1） articles in the field of neural regeneration; （2） fundamental research on animals, clinical trials and case reports; （3） article types： article, review, proceedings paper, note, letter, editorial material, discussion, book chapter; （4） year of publication： 2003-2012; and （5） citation database： Science Citation Index-Expanded. Exclusive criteria： （1） articles requiring manual searching or with access only by telephone; （2） unpublished articles; and （3） corrections. RESULTS： A total of 4 893 papers were retrieved from the Web of Science published between 2003 and 2012. The papers covered 65 countries or regions, of which the United States ranked first with 1 691 papers. The most relevant papers were in the neurosciences and cell biology, and the key- word ＂stem cell＂ was the most frequent. In recent years, China showed a great increase in the number of papers. Over the entire 10 years, there were 922 Chinese papers, with Jilin University ranking first with 58 articles. Chinese papers were published in connection with many countries, in- cluding the United States, Japan, and the United Kingdom. Among the connections, the papers published by the Chinese and the American are 107, with the highest rate. With regard to funding, 689 articles were funded from various projects, occupying 74.72% of the total amount. In these projects, National Foundation and Science and Technology programs were the majority. CONCLUSION： Our bibliometric analysis provides a historical perspective on the progress of neural regeneration research. At present, the number of articles addressing neural regeneration is increasing rapidly; however, through analysis of citations it is clear that there is a long way to go to improve the academic quality.

Effect of electrical stimulation on neural regeneration via the p38-RhoA and ERK1/2-Bcl-2 pathways in spinal cord-injured rats

Although electrical stimulation is therapeutically applied for neural regeneration in patients, it remains unclear how electrical stimulation exerts its effects at the molecular level on spinal cord injury （SCI）. To identify the signaling pathway involved in electrical stimulation improving the function of injured spinal cord, 21 female Sprague-Dawley rats were randomly assigned to three groups： control （no surgical intervention, n = 6）, SCI （SCI only, n = 5）, and electrical simulation （ES; SCI induction followed by ES treatment, n = 10）. A complete spinal cord transection was performed at the 10^th thoracic level. Electrical stimulation of the injured spinal cord region was applied for 4 hours per day for 7 days. On days 2 and 7 post SCI, the Touch-Test Sensory Evaluators and the Basso-Beattie-Bresnahan locomotor scale were used to evaluate rat sensory and motor function. Somatosensory-evoked potentials of the tibial nerve of a hind paw of the rat were measured to evaluate the electrophysiological function of injured spinal cord. Western blot analysis was performed to measure p38-RhoA and ERK1/2-Bcl-2 pathways related protein levels in the injured spinal cord. Rat sensory and motor functions were similar between SCI and ES groups. Com- pared with the SCI group, in the ES group, the latencies of the somatosensory-evoked potential of the tibial nerve of rats were significantly shortened, the amplitudes were significantly increased, RhoA protein level was significantly decreased, protein gene product 9.5 expression, ERK1/2, p38, and Bcl-2 protein levels in the spinal cord were significantly increased. These data suggest that ES can promote the recovery of electrophysiological function of the injured spinal cord through regulating p38-RhoA and ERK1/2-Bcl-2 pathway-related protein levels in the injured spinal cord.

Puerarin accelerates neural regeneration after sciatic nerve injury

Puerarin is a natural isoflavone isolated from plants of the genus Pueraria and functions as a protector against cerebral ischemia. We hypothesized that puerarin can be involved in the repair of peripheral nerve injuries. To test this hypothesis, doses of 10, 5, or 2.5 mg/kg per day puer- arin （8-（β-D-Glucopyranosyl-7-hydroxy-3-（4-hydroxyphenyl）-4H-l-benzopyran-4-one） were injected intraperitoneally into mouse models of sciatic nerve injury. Puerarin at the middle and high doses significantly up-regulated the expression of growth-associated protein 43 in the L4_6 segments of the spinal cord from mice at 1, 2, and 4 weeks after modeling, and reduced the atro- phy of the triceps surae on the affected side and promoted the regeneration of nerve fibers of the damaged spinal cord at 8 weeks after injury. We conclude that puerarin exerts an ongoing role to activate growth-associated protein 43 in the corresponding segment of the spinal cord after sciat- ic nerve injury, thus contributing to neural regeneration after sciatic nerve injuries.

Neural regeneration after peripheral nerve injury repair is a system remodelling process of interaction between nerves and terminal effector

In China, there are approximately 20 million people suffering from peripheral nerve injury and this number is increasing at a rate of 2 million per year. These patients cannot live or work independently and are a heavy responsibility on both family and society because of extreme disability and dysfunction caused by peripheral nerve injury （PNI）. Thus, repair of PNI has become a major public health issue in China.

Effect of Retinoic Acid on Expression of LINGO-1 and Neural Regeneration after Cerebral Ischemia

The purpose of this study was to observe the expression of LINGO-1 after cerebral ischemia, investigate the effects of retinoic acid（RA） on the expression of LINGO-1 and GAP-43, and the number of synapses, and to emplore the repressive effect of LINGO-1 on neural regeneration after cerebral ischemia. The model of permanent focal cerebral ischemia was established by the modified suture method of middle cerebral artery occlusion（MCAO） in Sprague-Dawley（SD） rats. The expression of LINGO-1 was detected by Western blotting and that of GAP-43 by immunohistochemistry. The number of synapses was observed by transmission electron microscopy. The SD rats were divided into three groups： sham operation（sham） group, cerebral ischemia（CI） group and RA treatment（RA） group. The results showed that the expression level of LINGO-1 at 7th day after MCAO in sham, CI and RA groups was 0.266±0.019, 1.215±0.063 and 0.702±0.081, respectively（P〈0.01）. The number of Gap-43-positive nerve cells at 7th day after MCAO in sham, CI and RA group was 0, 59.08±1.76 and 76.20±3.12 per high power field, respectively（P〈0.05）. The number of synapses at 7th day after MCAO was 8.42±0.13, 1.74±0.37 and 5.39±0.26 per μm2, respectively（P〈0.05）. It is concluded that LINGO-1 expression is up-regulated after cerebral ischemia, and RA inhibits the expression of LINGO-1, promotes the expression of GAP-43 and increases the number of synapses. It suggests that LINGO-1 may be involved in the pathogenesis of cerebral ischemia, which may provide an experimenal basis for LINGO-1 antogonist, RA, for the treatment of cerebral ischemia.

Ursolic acid induces neural regeneration after sciatic nerve injury

In this study, we aimed to explore the role of ursolic acid in the neural regeneration of the injured sciatic nerve. BALB/c mice were used to establish models of sciatic nerve injury through unilateral sciatic nerve complete transection and microscopic anastomosis at 0.5 cm below the ischial tuber-osity. The successful y generated model mice were treated with 10, 5, or 2.5 mg/kg ursolic acid via intraperitoneal injection. Enzyme-linked immunosorbent assay results showed that serum S100 protein expression level gradual y increased at 1-4 weeks after sciatic nerve injury, and significantly decreased at 8 weeks. As such, ursolic acid has the capacity to significantly increase S100 protein expression levels. Real-time quantitative PCR showed that S100 mRNA expression in the L 4-6 segments on the injury side was increased after ursolic acid treatment. In addition, the muscular mass index in the soleus muscle was also increased in mice treated with ursolic acid. Toluidine blue staining revealed that the quantity and average diameter of myelinated nerve fibers in the injured sciatic nerve were significantly increased after treatment with ursolic acid. 10 and 5 mg/kg of ursolic acid produced stronger effects than 2.5 mg/kg of ursolic acid. Our findings indicate that ursolic acid can dose-dependently increase S100 expression and promote neural regeneration in BALB/c mice fol owing sciatic nerve injury.

Rhesus monkey neural stem cell transplantation promotes neural regeneration in rats with hippocampal lesions

Rhesus monkey neural stem cells are capable of differentiating into neurons and glial cells. Therefore, neural stem cell transplantation can be used to promote functional recovery of the nervous system. Rhesus monkey neural stem cells （1 ×10^5 cells/μL） were injected into bilateral hippocampi of rats with hippocampal lesions. Confocal laser scanning microscopy demonstrated that green fluorescent protein-la- beled transplanted cells survived and grew well. Transplanted cells were detected at the lesion site, but also in the nerve fiber-rich region of the cerebral cortex and corpus callosum. Some transplanted cells differentiated into neurons and glial cells clustering along the ventricular wall, and integrated into the recipient brain. Behavioral tests revealed that spatial learning and memory ability improved, indicating that rhesus monkey neural stem cells noticeably improve spatial learning and memory abilities in rats with hippocampal lesions.

Translocator protein ligand, YL-IPA08, attenuates lipopolysaccharide-induced depression-like behavior by promoting neural regeneration

Translocator protein has received attention for its involvement in the pathogenesis of depression. This study assessed the effects of the new translocator protein ligand, YL-IPA08, on alleviating inflammation-induced depression-like behavior in mice and investigated its mechanism of action. Mice were intracerebroventricularly injected with 1, 10, 100 or 1000 ng lipopolysaccharide. The tail-suspension test and the forced swimming test confirmed that 100 ng lipopolysaccharide induced depression-like behavior. A mouse model was then established by intraventricular injection of 100 ng lipopolysaccharide. On days 16-24 after model establishment, mice were intragastrically administered 3 mg/kg YL-IPA08 daily. Immunohistochemistry was used to determine BrdU and NeuN expression in the hippocampus. YL-IPA08 effectively reversed the depression-like behavior of lipopolysaccharide-treated mice, restored body mass, increased the number of BrdU-positive cells, and the number and proportion of BrdU and NeuN double-positive cells. These findings indicate that YL-IPA08 can attenuate lipopolysaccharide-induced depression-like behavior in mice by promoting the formation of hippocampal neurons.

Roles of neural regeneration in memory pharmacology

Neural regeneration, or neuroregeneration, is a brain mechanism essential for rescuing cognitive functions pharmacologically against memory disorders and aging-related memory abnormality. In this short Perspective article, we intend to briefly present the essential roles of neuroregeneration and neural plasticity in learning and memory, memory disorders, and critical involvement in an effective treatment of memory disorders.

Synthetic and nature-derived lipid nanoparticles for neural regeneration

Challenges and opportunities in nerve regeneration： The central nervous system （CNS） has a limited ability to regen- erate. Subsequent to spinal injury, glial scar formation, creat- ed by fibroblasts, neuroglia, monocytes, and endothelial cells, inhibits regeneration of the injured nerve. The peripheral nervous system （PNS） has a greater regeneration potential than the CNS; however, the current gold standard of treat- ment for a large nerve defect is still autologous nerve grafts, which require multiple surgeries. For this reason, researchers have been trying to regenerate nervous tissues, including brain, spinal cord. and PeriPheral nerves, for decades.

Efficacy of glucagon-like peptide-1 mimetics for neural regeneration

Glucagon-like peptide 1（GLP-1）is secreted from enteroendocrine L cells in response to nutrient ingestion and exhibits insulinotropic properties by stimulating specific G protein-linked receptors（GLP-1Rs）on pancreaticβcells.Several GLP-1 mimetics,such as exenatide（exendin-4（Ex-4））,liraglutide,and lixisenatide,have been developed and approved as treatments for patients with type 2 diabetes.These peptides show bioactiv-ities almost identical to those of GLP- 1 and have a substantially longer plasma half-life than GLP-1 because of their resistance to dipeptidyl peptidase-4, a GLP-1 degrading enzyme.

Maggot homogenate is associated with neural regeneration and wound healing in the rat

BACKGROUND：Live delivery limits the clinical application of maggot therapy. To date in China, there are no in vivo reports regarding wound healing mechanisms of maggot therapy or the effects of maggot homogenate on wound nerve regeneration.OBJECTIVE：To avoid complications due to the use of live maggots, an aseptic maggot homogenate was applied. Substance P （SP） and gene protein product 9.5 expression in a cutaneous wound was analyzed to explore possible mechanisms of neural regeneration and wound healing in the rat.DESIGN, TIME AND SETTING：A random grouping and controlled animal study was performed at the laboratory of the Department of Orthopedic Surgery, First Affiliated Hospital, Dalian Medical University from August 2008 to April 2009.MATERIALS：Live maggots were cultured and provided by the laboratory of the Department of Orthopedic Surgery of the First Affiliated Hospital, Dalian Medical University, China.METHODS：A total of 48 adult rats were selected and two acute, full-thickness wounds （round, 1.5 cm diameter） were created on the back of each rat. The two wounds were randomly assigned to homogenate product and control groups. Following two-step disinfection of maggots, a homogenate was produced from 10 maggots and applied to the wound area in the homogenate product group, while the wounds in the control group were treated with normal saline alone.MAIN OUTCOME MEASURES：On days 1,3, 7, 10, 14, and 21 following injury, the wound tissue was excised. Histological examination of the wound was observed by hematoxylin and eosin staining or Masson＇s Trichrome staining. SP and protein gene product 9.5 expressions were examined by immunohistochemistry to evaluate wound neural regeneration.RESULTS：On days 7, 10, and 14, the rate of wound healing was significantly greater in the homogenate product group compared with the control group （P 〈 0.05）, and homogenate healing was better than that seen in the control group. On days 3, 7, and 10, SP expression in cells and regenerative nerves was significantly greater in the homogenate product group compared with the control group （P 〈 0.05）. On days 7 and 10, protein gene product 9.5 expression was detected in the regenerative nerve, and expression level was significantly greater in the homogenate product group compared with the control group （P 〈 0.05）.CONCLUSION：Maggot homogenate resulted in upregulated SP and protein gene product 9.5 expressions, thereby promoting neural regeneration and wound healing.

National Institutes of Health Funding for disease categories related to Neural Regeneration Research

The National Institutes of Health （NIH）, a part of the U.S. Department of Health and Human Services, is the nation＇s medical research agency-making important discoveries that improve health and save lives.Thanks in large part to NIH-funded medical research, Americans today are living longer and healthier. Life expectancy in the United States has jumped from 47 years in 1900 to 78 years as reported in 2009, and disability in people over age 65 has dropped dramatically in the past 3 decades. In recent years, nationwide rates of new diagnoses and deaths from all cancers combined have fallen significantly.

Latest data on articles from Neural Regeneration Research (NRR) indexed by SCI,Chemical Abstracts (CA) and Excerpta Medica (EM)

NRR retrieved articles indexed by SCI, CA and EM at the beginning of March, 2008. The most recent data are as follows：