Method for treating syringomyelia by using stem cells and application

Abstract

The invention discloses a method for treating syringomyelia by using stem cells and application, and provides application of the stem cells in preparation of a medicine, and the medicine can be used for treating or preventing at least one of the following diseases: 1) the medicine is used for treating or adjunctively treating syringomyelia; 2) being used for treating nerve injury or nerve protection after syringomyelia; 3) being used for promoting the repair of neuralgia after syringomyelia; 4) for preventing syringomyelitis or preventing the further development of syringomyelitis; 5) preventing relapse of syringomyelia; and 6) resisting inflammation. Through cell transplantation, hole shrinkage can be promoted, a local inflammation environment can be adjusted, and a novel method with a wide prospect is provided for treating syringomyelia.

Description

Technical Field

[0001] The present invention belongs to the technical field of stem cells. Specifically, it relates to a method and application for treating syringomyelia using stem cells, especially using mesenchymal stem cells and neural stem cells to treat syringomyelia characterized by focal dilation of the central canal. Background Art

[0002] Syringomyelia is a chronic progressive disease, which refers to a longitudinal cavity filled with fluid in the spinal cord. The pathogenesis of syringomyelia is mainly caused by Chiari malformation, but it can also be caused by spinal cord injury, tumors, myelitis, arachnoiditis, spinal cord tethering and other factors. Acquired syringomyelia is mainly due to the disruption of the normal cerebrospinal fluid (CSF) circulation. However, the pathogenesis of acquired syringomyelia remains controversial.

[0003] Syringomyelia can lead to the gradual loss of function in patients, especially the loss of mobility, and usually requires surgical intervention. Although the progression of dysfunction can be stabilized after intervention, the symptoms usually persist. Especially for central pain syndrome, despite adequate surgical treatment and the disappearance of the syrinx shown on imaging, there is still a great deal of treatment resistance.

[0004] The potential clinical pathogenesis of syringomyelia leading to nerve pain and sensory disturbances remains unclear. Due to possible nerve injury or a chronic inflammatory environment associated with syringomyelia, the symptoms may not be relieved even after syringomyelia decompression surgery. First, studies have shown that the basic pathology of syringomyelia is a gradually expanding cavity within the central canal. This "syrinx" filled with cerebrospinal fluid continuously expands, compressing the nerve fibers of the anterior white commissure, resulting in the loss of pain and temperature sensation, while touch and vibration sensation are preserved (segmental dissociated sensory loss). Therefore, spinal cord injury caused by the compression of the enlarged cavity is one of the causes of the disease symptoms. Eliminating the pressure caused by syringomyelia by reducing the "syrinx" is of great significance for recovery. Second, an adverse environment can also lead to further nerve damage, hindering nerve regeneration and repair.

[0005] Cell therapy is expected to become a major method for treating syringomyelia caused by spinal cord injury and trauma. Mesenchymal stem cells are widely used due to their immunomodulatory ability and the ability to bypass ethical barriers related to stem cell transplantation. It has been proven that mesenchymal stem cells play a key role in tissue repair and microenvironment regulation in the treatment of spinal cord injury. INSCs (induced neural stem cells) are the main cell type in current research on neural stem cell treatment of spinal cord injury, and neural stem cells are also involved in cell-mediated regeneration and plasticity after spinal cord injury.

[0006] However, there has been little research on the treatment of FDCC-syringomyelia with cell therapy. In 2022, a case report stated that mesenchymal stem cells could effectively treat such syringomyelia, but there was a lack of direct evidence between the cells and syringomyelia. As for the application of neural stem cells, there have been no reports on the application of neural stem cells to such syringomyelia. In addition, there has been no systematic research on cell transplantation methods. Therefore, there is an urgent need for an effective method for treating FDCC-syringomyelia at present. Summary of the Invention

[0007] The present invention aims to provide a method and application for treating syringomyelia using stem cells. Through cell transplantation, it can promote the reduction of the syrinx and regulate the local inflammatory environment, providing a new method with broad prospects for treating syringomyelia.

[0008] On the one hand, the present invention provides an application of stem cells in the preparation of a drug, and the drug can be used for treating or preventing at least one of the following diseases:

[0009] 1) For treating or adjuvant-treating syringomyelia;

[0010] 2) For treating nerve injury after syringomyelia or for neuroprotection;

[0011] 3) For promoting the recovery of neuralgia after syringomyelia;

[0012] 4) For preventing syringomyelia or preventing the further development of syringomyelia;

[0013] 5) For preventing the recurrence of syringomyelia;

[0014] 6) For anti-inflammation.

[0015] In one embodiment of the present invention, the syringomyelia is of the central canal dilation type.

[0016] In one embodiment of the present invention, the treatment is sufficient to detectably improve one or more of the following symptoms: reduction of the syrinx, increase in endogenous stem cells, increase in ependymal cells, increase in M2-phenotype microglia, improvement of sensory disturbances caused by syringomyelia, and improvement of inflammation caused by syringomyelia.

[0017] In one embodiment of the present invention, the stem cells are derived from umbilical cord blood, peripheral blood, bone marrow, or brain tissue.

[0018] In one embodiment of the present invention, the stem cells include mesenchymal stem cells and / or neural stem cells.

[0019] In one embodiment of the present invention, the treatment or prevention includes administering stem cells into the body of an object.

[0020] In one embodiment of the present invention, the administration site includes syringomyelia, subarachnoid space, and spinal cord parenchyma.

[0021] In a preferred embodiment of the present invention, the administration site is syringomyelia.

[0022] In one embodiment of the present invention, the administration site is located by MRI.

[0023] In one embodiment of the present invention, the administration method includes injection.

[0024] In one embodiment of the present invention, the stem cells are administered at a therapeutic dose.

[0025] In one embodiment of the present invention, the therapeutic dose of the stem cells is 0.5 - 1×10 5 cells / μL.

[0026] In one embodiment of the present invention, the administration frequency is once a day or once every two days.

[0027] In one embodiment of the present invention, the subject is a mammal.

[0028] In one embodiment of the present invention, the subject is a human.

[0029] Use of a composition in the preparation of a drug, the drug having at least one of the following therapeutic effects:

[0030] 1) For treating or assisting in the treatment of syringomyelia;

[0031] 2) For treating nerve injury after syringomyelia or for nerve protection;

[0032] 3) For promoting the repair of neuralgia after syringomyelia;

[0033] 4) For preventing syringomyelia or preventing the further development of syringomyelia;

[0034] 5) For preventing the recurrence of syringomyelia;

[0035] 6) For anti - inflammation.

[0036] In one embodiment of the present invention, the composition includes the above - mentioned stem cells.

[0037] In one embodiment of the present invention, the composition further includes other drugs for treating syringomyelia.

[0038] In one embodiment of the present invention, the other drugs include neurotrophic drugs.

[0039] In a specific embodiment of the present invention, the neurotrophic drugs include, but are not limited to, vitamin B1, adenosylcobalamin, and mecobalamin.

[0040] The application of a preparation in the preparation of a drug, wherein the drug has at least one of the following therapeutic effects:

[0041] 1) For the treatment or adjuvant treatment of syringomyelia;

[0042] 2) For the treatment of nerve injury or nerve protection after syringomyelia;

[0043] 3) For promoting the repair of neuropathic pain after syringomyelia;

[0044] 4) For preventing syringomyelia or preventing the further development of syringomyelia;

[0045] 5) For preventing the recurrence of syringomyelia;

[0046] 6) For anti - inflammation.

[0047] In an embodiment of the present invention, the preparation includes the above - mentioned stem cells.

[0048] In an embodiment of the present invention, the preparation further includes a pharmaceutically acceptable carrier or excipient.

[0049] Advantages of the present invention:

[0050] 1) The present invention uses mesenchymal stem cells and neural stem cells to treat syringomyelia characterized by focal dilation of the central canal. Using a rat model of syringomyelia characterized by focal dilation of the central canal (FDCC), an effective transplantation method was first determined using iNSCs, and then the effects of iNSCs and mesenchymal stem cells on syringomyelia were compared. The experimental results show that effective transplantation of mesenchymal stem cells (MSCs) and induced neural stem cells (iNSCs) can treat syringomyelia. The present invention provides research evidence for cell therapy for FDCC - syringomyelia.

[0051] 2) The present invention discovers for the first time that cell transplantation can not only promote cavity shrinkage, but also stimulate the proliferation of ependymal cells, and its effect is related to the transplantation location. These results are of great significance for the activation of endogenous stem cells and nerve regeneration after syringomyelia.

[0052] 3) The present invention also discovers that cell transplantation transforms activated microglia into the M2 phenotype, and M2 microglia expressing IGF1 may play an important role in the repair of neuropathic pain.

[0053] 4) The present invention also found that stem cell transplantation can significantly reduce the cavity and improve the microenvironment, highly suggesting that cell therapy can improve the sensory disorders caused by the cavity. If stem cell therapy can be used simultaneously through surgery, it may be an effective method to relieve the symptoms caused by the sensory pathway.

[0054] In summary, the present invention uses cell transplantation to promote cavity reduction and regulate the local inflammatory environment, and the proliferation of ependymal cells indicates that endogenous stem cells are activated, which is very important for the regeneration and repair of spinal cord injury and has the potential to relieve neuropathic pain and improve sensory disorders, providing a new method with broad prospects for the treatment of syringomyelia. Brief Description of the Drawings

[0055] Figure 1 It is a diagram showing the influence of the transplantation site on the results. A and B are the transplantation sites (X = 0); C - E are the MRI images of the spinal cords of three groups of rats before and after cell transplantation; F is the percentage of spinal cord cavity shrinkage in the three groups (n = 3; *P < 0.05; **P < 0.01; ***P < 0.001);

[0056] Figure 2 It is a diagram showing the histological staining results of cells transplanted into the spinal cord cavity. A is the control group; B is INSC transplanted into the spinal cord parenchyma; C is INSC transplanted into the cavity. The cavity shrinks, and iNSC can be seen in the cavity. (Red arrows indicate the cell transplantation site; Bar = 0.5 mm)

[0057] Figure 3 is a diagram showing the results after mesenchymal stem cells and iNSCs are transplanted into the spinal cord cavity; A is the MRI image of the spinal cord of a control group rat before and after transplantation and the immunofluorescence staining of the corresponding spinal cord section of the rat. It can be seen that there is a persistent large - area spinal cord cavity in the rat spinal cord; B is mesenchymal stem cells transplanted into the spinal cord cavity. The MRI image and immunofluorescence results show that the spinal cord cavity of the same rat shrinks; C is iNSCs seen in the spinal cord cavity, and the spinal cord cavity shrinks. C1 - C3 are the magnified views of the corresponding positions of iNSC in consecutive sections (Hu - Nu, human nuclear antigen is red, GFP is green). D is the proportion of cavity shrinkage in the three groups. Bar = 200 μm (groups A - C); Bar = 100 μm (groups C1 - C3) (n = 3; *P < 0.05; **P < 0.01; ***P < 0.001);

[0058] Figure 4 is a diagram showing the influence of cell transplantation on ependymal cells. A - D are the central canal morphologies of the carrier group, iNSCs - parenchyma group, iNSCs - cavity group, and MSCs - cavity group; Sox2 - positive cells are ependymal cells. E is the integrated optical density of Sox2 - positive ependymal cells per unit length (n = 6; *P < 0.05; **P < 0.01;

[0059] ***P < 0.001; Bar = 200 μm);

[0060] Figure 5 shows the expression results of Nestin+ cells after cell transplantation; A shows the expression of Nestin in the central canal of rats in different groups; the number of Nestin-positive cells increased in the vector group, iNSCs-cavity group, and MSCs-cavity group; B shows the comparison of Nestin+ cells under four different conditions.

[0061] Bar = 50 μm (n = 3; *P < 0.05; **P < 0.01; ***P < 0.001).

[0062] Figure 6 shows the activation results of Iba1-positive microglia around the spinal cord cavity; A shows the activation results of microglia after transplantation in normal rats, control group, iNSCs-cavity group, and MSCs-cavity group; B shows the quantitative analysis of microglia proliferation after transplantation of iNSCs and mesenchymal stem cells in different groups (in the area within 0.5 mm around the spinal cord cavity) (n = 6; Bar = 200 μm; *P < 0.05; **P < 0.01; ***P < 0.001);

[0063] Figure 7 shows the results of microglia expressing CD206 and TNF-α; A and B show that after transplantation of mesenchymal stem cells and iNSCs, CD206 and TNFα were expressed, but microglia rarely expressed CD206 and TNFα; C shows that no microglia in the vector group expressed CD206 and TNF-α. (Yellow arrows indicate microglia expressing TNFα or CD206)

[0064] Figure 8 shows the expression results of M2-type microglia. A shows that the activated microglia in the iNSCs-cavity group and MSCs-cavity group mainly expressed IGF1 (yellow arrows indicate microglia expressing IGF1). B shows the quantitative analysis of IGF1 in different groups. The expression of IGF1 increased in the iNSCs-cavity group and MSCs-cavity group (n = 6; *P < 0.05; **P < 0.01; ***P < 0.001); Detailed implementation manners

[0065] In order to make the objectives, technical solutions, and advantages of the present invention clearer and more understandable, the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be emphasized that the specific embodiments described herein are only used to better illustrate the present invention, which are partial embodiments of the present invention, rather than all embodiments, so they are not used to limit the present invention. In addition, the technical features involved in the embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0066] The materials, reagents, etc. used in the following examples can be obtained from commercial sources unless otherwise specified.

[0067] The detection methods and analysis methods used in the following examples are as follows:

[0068] Method for magnetic resonance imaging of rats:

[0069] Magnetic resonance imaging tests were performed using a 7.0 Tesla nuclear magnetic resonance imaging scanner (PharmaScan 7T, Bruker, Karlsruhe, Germany) with a gradient of 400 mT / m. The rats were placed on a table and their trunks were fixed with two restraint straps. After a rapid whole-body localization scan, sagittal and axial T2-weighted images were acquired using a fat-saturated RARE sequence with the surgical site as the center. A rat volume coil with a diameter of 89 mm was used for transmission and data acquisition. Data analysis was performed using RadiAnt DICOM Viewer software (version 4.6.9, Medixant, Poznan, Poland).

[0070] Immunohistochemical analysis method:

[0071] Two weeks after cell transplantation, all rats were perfused transcardially with 0.9% sodium chloride solution and then treated with 4% paraformaldehyde (PFA). The spinal cords were post-fixed, immersed in 30% sucrose solution at 4°C for 48 hours, and embedded in OCT. The spinal cord segments containing cavities were frozen and stored at -80°C, or cut into 20-μm-thick sections using a cryomicrotome. Immunofluorescence was performed using the following primary antibodies: mouse anti-green fluorescent protein (GFP, 1:500, eBiosciense), goat anti-ionized calcium-binding adaptor molecule-1 (Iba1, 1:500, Abcam), mouse anti-GFAP (1:500, Santa Cruz), rabbit anti-GFAP (1:200, Zhongshan Jinqiao), mouse anti-SOX2 (1:400, Santa Cruz), rabbit anti-SOX2 (1:400, CST), mouse anti-nestin (1:400, Millipore), mouse anti-human nucleus (Hu-Nu, 1: rabbit anti-KI67 (1:500, Millipore), rabbit anti-CD206 (1:400, Abclonal), rabbit anti-TNF-α (1:400, Abclonal). The primary antibodies were diluted in 0.01 M phosphate-buffered saline (PBST) containing 0.3% Triton X-100. The secondary antibodies were conjugated with Alexa Fluor 488 (1:200, Jackson), Cy3 (1:200, Jackson), or Alexa Fluor 647 (1:200, Jackson). All sections were pre-incubated in PBST containing 3% donkey serum albumin for 30 minutes, incubated with the primary antibodies overnight at 4°C, rinsed, and then incubated with the secondary antibodies for 1 hour at room temperature. Finally, all sections were counterstained with the nuclear marker DAPI (1 μg / mL, YEASEN). Images were acquired using a laser scanning confocal microscope Leica SCN400 (Leica Microsystems).

[0072] The method for measuring spinal cord cavities is as follows:

[0073] Measure the diameters of all cavities observed in the magnetic resonance imaging results. The sum of the diameters is the size of the spinal cord cavity. The percentage of cavity reduction is the ratio of the cavity reduction value after transplantation to the cavity size before transplantation. The statistical analysis results are expressed as mean ± SEM (standard error of the mean, SEM). Statistical analysis was performed using SPSS 20 software. The Gaussian distribution of the data set was evaluated using the one-sample Kolmogorov-Smirnov test. One-way analysis of variance was used to determine the differences between multiple groups. P < 0.05 was considered statistically significant.

[0074] Example 1

[0075] Establishment of an animal model of spinal cord cavity

[0076] 1. Selection of animals

[0077] Twenty adult female Sprague-Dawley rats (provided by Vital River Laboratories, Beijing, China), weighing between 220 - 250 grams, were used to establish a syringomyelia model. All rats were housed under standard conditions in the animal experiment center of Xuanwu Hospital. The animal experiments were approved by the Animal Ethics Committee of Xuanwu Hospital (XW-20210723-1) and complied with the "Regulations on the Administration of Animals in China".

[0078] 2. Establishment of an animal model of syringomyelia

[0079] Under anesthesia, a 3-cm incision was made in the midline of the rat's back. The paravertebral muscles and the intervertebral tissue between T12 and T13 were dissected under a microscope to expose the T12 / T13 intervertebral space and the ligamentum flavum. Then, the ligamentum flavum was carefully cut with a coronary scissors. A cotton wick was twisted into a thin strip and inserted into the epidural space under the T13 lamina. After flushing the surgical area with normal saline, the muscles and skin were sutured. Penicillin was injected intraperitoneally to prevent postoperative infection. Alternatively, a syringomyelia rat model can also be established by referring to the literature (Fluids Barriers CNS. 2020 Jul 31; 17(1): 50.).

[0080] In vivo magnetic resonance imaging was performed on the rats. The results showed that 15 rats exhibited central canal dilation in the magnetic resonance imaging results, with the maximum diameter of the central canal dilation being greater than 0.5 mm, meeting the inclusion criteria and being included in the experimental group.

[0081] Example 2

[0082] Preparation of stem cells

[0083] Mesenchymal stem cells were extracted from the bone marrow of 4-week-old male juvenile SD rats and cultured to passage P1. Then they were frozen and stored at -80 °C for later use. Before the experiment, the passage P1 mesenchymal stem cells were thawed and transfected with lentivirus carrying FUGW (GFP) when the cell proliferation reached 80%. The cells were labeled with GFP and cultured to passage P2 for transplantation. Neural stem cells were isolated from the brain tissue of neonatal SD suckling rats in the same way. The mesenchymal stem cell (MSCs) suspension contained 5×10 4 cells / μL, and the neural stem cell (iNSCs) suspension contained 1×10 5 cells / μL.

[0084] Example 3

[0085] Decompression surgery and cell transplantation

[0086] 1. Decompression surgery

[0087] Six weeks after surgery, the syringomyelia rats were decompressed. The rats were anesthetized with enflurane, and then the lamina and vertebral bars were removed. The cotton was carefully peeled off from the compression site without damaging the spinal cord.

[0088] 2. Cell transplantation

[0089] MRI locates the spinal cord cavity, and the cell transplantation site is determined according to the location of the cavity. Figure 1 As shown in A and B, the compression edge (coracoid process) and the midpoint of the spinal cord are taken as the origin (X=0), the distance from the spinal cord cavity to the origin is Y, and the depth is Z. A total volume of 10 μL of cell suspension was transplanted into each rat using a microsyringe (Agilent, 10 μL) under a stereomicroscope (RWD). Starting 24 hours before transplantation, all rats were subcutaneously injected with 10 mg / kg cyclosporine (Sandimmun, Novartis) once a day for immunosuppression. The experiment included four transplantation groups: iNSCs transplantation into the cavity group (iNSCs-cavity group, 3 rats), iNSCs transplantation into the spinal cord parenchyma group (iNSCs-parenchyma group, 3 rats), mesenchymal stem cell transplantation into the cavity group (MSCs-cavity group, 3 rats) and vehicle transplantation into the cavity group (vehicle group, 6 rats). Cell transplantation was performed on the four groups of rats.

[0090] Example 4

[0091] Relationship between the reduction of syringomyelia and the location of transplanted cells

[0092] The iNSCs suspension (iNSCs-syrinx group) and the vector (vector group / control group) were transplanted into the largest syrinx of the spinal cord, and the iNSCs (iNSCs-parenchyma group) were transplanted into the spinal cord parenchyma. Figure 1 A and Figure 1 B. The size of the cavity was detected by magnetic resonance imaging, and the results were as follows Figure 1 The results showed that after iNSCs were transplanted into the cavity, the central lumen was reduced ( Figure 1 C and 1D), while the central lumen of the control group was not completely contracted ( Figure 1 E). The syringomyelia of the iNSCs-syringomyelia group shrank by 87.4%±21.82%, the syringomyelia of the iNSCs-parenchyma group shrank by 11.6%±9.8%, and the syringomyelia of the control group shrank by 36.68%±27.71% ( Figure 1 F). These results were consistent with the results of tissue staining, which showed that the dilated central canal was significantly reduced in the iNSCs-cavity group compared with the other two groups ( Figure 2 Although simple decompression can lead to the reduction of the syringomyelia without cell intervention, the effect is not as good as transplanting cells into the syringomyelia.

[0093] It can be seen that cell transplantation plays a crucial role in syringomyelia reduction and depends on the transplantation location.

[0094] Example 5

[0095] Effects of Mesenchymal Stem Cells and Neural Stem Cells on Syringomyelia

[0096] Mesenchymal stem cells (MSCs-syringomyelia group) and a carrier were transplanted into the syringomyelia cavity. A total of six rats were used, three of which were transplanted with mesenchymal stem cells and three with the carrier. The effects of mesenchymal stem cells and neural stem cells (same as Example 4) on syringomyelia were compared. The size of the syringomyelia was detected by magnetic resonance imaging. The results are shown in Figure 3. The results showed that after transplantation of mesenchymal stem cells and iNSCs into the syringomyelia cavity, the syringomyelia significantly shrank, and immunofluorescence staining also showed that the syringomyelia in the MSCs-syringomyelia group and (iNSCs-syringomyelia group) shrank ( Figures 3A-C ). After statistical analysis, there was no significant difference in the effects of MSCs and iNSCs on syringomyelia reduction ( Figure 3D ). A small number of GFP-labeled iNSCs expressing anti-human nucleoprotein were found in the central canal (Figure 3C1-C3). However, no surviving iNSCs were found in the spinal cord tissue. The results indicate that both types of cells can cause syringomyelia reduction with no difference in effect.

[0097] Example 6

[0098] Proliferation of Ependymal Cells after Cell Transplantation into the Central Canal

[0099] Sox2+ ependymal cells showed the location of the central canal (Figure 4A-4D). After cell transplantation into the syringomyelia cavity, the ependymal cells thickened, indicating ependymal cell proliferation. By comparing the expression of SOX2+ cells under different conditions, it was found that after transplantation of mesenchymal stem cells and iNSCs into the syringomyelia cavity, the number of SOX2-positive ependymal cells increased compared to when no cells were transplanted ( Figure 4E ). Rats without iNSC transplantation showed persistent large-area syringomyelia and no obvious ependymal cell proliferation (Figure 4B and 4E). The enhanced ependymal cell proliferation indicates that cell transplantation may activate endogenous stem cells.

[0100] Staining was performed for the stem cell marker nestin. Two weeks after transplantation, it was observed that the number of nestin-positive cells in the MSCs-syringomyelia group and iNSCs-syringomyelia group increased compared to the carrier group and negative control group (non-transplanted group), and there was no difference between the MSCs-syringomyelia group and iNSCs-syringomyelia group (Figure 5). These results further confirm that cell transplantation into the syringomyelia cavity may promote the activation of endogenous stem cells.

[0101] KI67-positive cells were not detected in the proliferating ependymal cells, indicating that these cells did not continue to proliferate at this time, thus reducing the tumor risk (results not shown).

[0102] Example 7

[0103] Activated microglia polarized into the M2 phenotype after cell transplantation

[0104] Under physiological conditions, microglia have a branched morphology and are distributed outside the ependymal cells. However, when syringomyelia forms, microglia are activated, resulting in an enlarged cell body and a spherical or rod-shaped morphology ( Figure 6A ). Due to the dilation of the central canal, the tight junctions between ependymal cells are disrupted, resulting in an increased cell spacing, and microglia invade the ependymal cell layer. After cell transplantation, although syringomyelia was restored and ependymal cell proliferation was promoted, microglia still remained and the number increased. In the MSCs-syringomyelia group and the iNSCs-syringomyelia group, the expression level of microglia was higher, mainly concentrated at the cell transplantation site or around the ependymal cells two weeks after transplantation ( Figure 6B ).

[0105] Microglia have two main phenotypes, namely M1 and M2. M1 microglia are associated with tissue damage and inflammation, and recruit inflammatory cells by expressing cytokines such as tumor necrosis factor (TNF)-α and interleukin (IL)-1β. In contrast, M2 microglia can regulate tissue repair and have anti-inflammatory effects. M2-type microglia express IL-4, IL-10, CD206, and growth factors such as insulin-like growth factor 1 (IGF1) and transforming growth factor (TGF)-β.

[0106] The experimental results showed that few microglia expressed TNF-α and CD206 ( Figure 7 ), and activated microglia mainly expressed IGF1 ( Figure 8A ), indicating that they polarized into the M2 phenotype, and the M2 phenotype can promote nerve repair and have anti-inflammatory properties. After cell transplantation, especially IGF1 in the MSCs-syringomyelia group increased significantly ( Figure 8B ).

[0107] Thus, under the action of mesenchymal stem cells, cell transplantation may have a beneficial effect on the recovery of neuropathic pain after syringomyelia.

[0108] Mechanism explanation: The location and function of ependymal cells in the ventricular system of the central nervous system make them an important cellular barrier for regulating molecular transport and exchange between the brain and the body. Some studies have pointed out that both the blood-cerebrospinal fluid barrier and the cerebrospinal fluid-brain barrier originate from ependymal cells, and their main function is to regulate the balance and dynamics of cerebrospinal fluid. Syringomyelia can lead to the dilation of the central canal, thereby disrupting the arrangement and tight junctions between individual ependymal cells, severely damaging the barrier function, and also causing neurodegeneration. After cell transplantation, the ependymal cell layer thickens, which may promote the proliferation of endogenous stem cells. This thickening may play a key role in establishing a stable barrier function, preventing further expansion of the cavity, and subsequent nerve damage.

[0109] The microenvironment around endogenous neural stem cells is a key factor in stem cell-mediated nerve repair. Studies have shown that the proliferation of endogenous stem cells is necessary to prevent further expansion of injuries, provide neurotrophic support, and has an impact on immune regulation and myelin regeneration. Therefore, the proliferation of ependymal cells and the increase in the expression of nestin-positive cells indicate an enhanced activation potential of endogenous stem cells, which may help improve the inflammatory environment at the syringomyelia site. At the same time, it is beneficial for tissue repair, nerve regeneration, and the treatment of sensory disorders related to syringomyelia. The present invention emphasizes the importance of precise positioning of cell transplantation to mobilize stem cells, thereby avoiding occupying normal tissues.

[0110] After decompression and cell transplantation, microglia are activated and proliferate. This activation may be due to the presence of exogenous grafts and the stress response after decompression. Some studies have shown that microglia are highly dynamic, undergo extensive proliferation within the first two weeks, and aggregate around the lesion. However, transplanted cells can affect the transformation of activated microglia into M2 cells, initiating a neuroprotective function. At this stage, microglia present around the syringomyelia and transplanted cells may play an important role in regulating the local microenvironment of syringomyelia, promoting the recovery of syringomyelia, and alleviating neuralgia.

[0111] The above are only the preferred application embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the technical principles of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. Use of a stem cell in the preparation of a drug, the drug being used for treating or preventing at least one of the following diseases: 1) for treating or adjuvant-treating syringomyelia; 2) for treating nerve injury after syringomyelia or for nerve protection; 3) for promoting the recovery of neuralgia after syringomyelia; 4) for preventing syringomyelia or preventing the further development of syringomyelia; 5) for preventing the recurrence of syringomyelia; 6) for anti-inflammation.

2. The use according to claim 1, wherein, The syringomyelia is of the central canal dilatation type of the spinal cord.

3. The use according to claim 1, wherein, The treatment is sufficient to detectably improve one or more of the following symptoms: cavity shrinkage, increased endogenous stem cells, increased ependymal cells, increased M2 phenotype microglia, improvement of sensory disturbances caused by syringomyelia, and improvement of inflammation caused by syringomyelia.

4. The use according to claim 1, wherein, The stem cells are derived from umbilical cord blood, peripheral blood, bone marrow or brain tissue.

5. The use according to claim 1, wherein, The stem cells include mesenchymal stem cells and / or neural stem cells.

6. The use according to claim 1, wherein, The treatment or prevention includes administering stem cells into the body of a subject; Preferably, the administration sites include the syringomyelia cavity, the subarachnoid space and the spinal cord parenchyma; More preferably, the administration site is the syringomyelia cavity; Preferably, magnetic resonance imaging (MRI) is used to locate the administration site.

7. The use according to claim 6, wherein, The stem cells are administered at a therapeutic dose.

8. The use according to claim 6, wherein, The administration frequency is once a day or once every two days.

9. Use of a composition in the preparation of a drug, the drug having at least one of the following therapeutic effects: 1) for treating or adjuvant-treating syringomyelia; 2) for treating nerve injury after syringomyelia or for nerve protection; 3) for promoting the repair of neuralgia after syringomyelia; 4) for preventing syringomyelia or preventing the further development of syringomyelia; 5) for preventing the recurrence of syringomyelia; 6) for anti-inflammation; wherein, The composition includes the stem cells described in claims 1-8; Preferably, the composition further includes other drugs for treating syringomyelia; Preferably, the other drugs include neurotrophic drugs; Preferably, the neurotrophic drugs include vitamin B1, adenosylcobalamin, and mecobalamin.

10. Use of a preparation in the preparation of a drug, wherein the drug has at least one of the following therapeutic effects: 1) for treating or assisting in the treatment of syringomyelia; 2) for treating or protecting nerves damaged after syringomyelia; 3) for promoting the repair of neuralgia after syringomyelia; 4) for preventing syringomyelia or preventing the further development of syringomyelia; 5) for preventing the recurrence of syringomyelia; 6) for anti - inflammation; wherein, The preparation includes the stem cells described in claims 1-8; Preferably, the preparation further includes a pharmaceutically acceptable carrier or excipient.

Images