Effect of Huangqi On Mesenchymal Stem Cells (MSC) Transplanted Therapy in Spinal Cord Injury: It Promotes the Neural Differentiation of MSC In Vivo.

Abstract Abstract 4476 Introduction The therapeutic effects of using mesenchymal stem cells (MSCs) for transplantation in the treatment of spinal cord injury have been previously studied. Astragalus propinquus, also known as Huangqi, is a traditional Chinese herb commonly used to strengthen the immune system and protect neuron. However, the possibility of combining the beneficial effects of both MSC transplantation and Huangqi Injecta in treating spinal cord injury has not been addressed. Methods A total of 120 male, four-week-old, Wistar rats, were randomly divided into six groups: sham group, model group, phosphate buffered saline (PBS) solution control group (PBS group), Huangqi injection control group (Huangqi group), rMSCs transplantation group (rMSCs group), rMSCs and Huangqi injecta treated group (rMSCs + Huangqi group). There were 10 rats in the sham group and 22 rats in the other groups. In the PBS group, rats were injected 5 μl PBS into the cephalic site of spinal cord injured section three days after the operation; in the Huangqi group, rats were intraperitoneally injected with Huangqi Injecta at a dose of 1.2 ml per 100 g of body weight on day three after the operation; in the rMSCs group, 5 μl of rMSC suspension (2.5–5×105 cells) was injected into the cephalic site of the spinal cord injured part in rats on day three after the operation; in the rMSCs + Huangqi group, rats were injected Huangqi Injecta and 5 μl rMSC suspension (2.5–5×105 cells) following the methods mentioned in the Huangqi group and the rMSCs group. Six animals were randomly picked from each groups on day 7, 14, 21 and 28 to receive Basso-Beattie-Bresnahan (BBB) scale grading for motor function checking of the hind legs. HE staining is adopt for the histopathologic examination, and Dual-marked immunohistochemical analysis is applied for cell locating and differentiation determination. Results The average BBB score in the Huangqi group is higher than model group and PBS group, except on day 21. The score in rMSCs group and the rMSCs + Huangqi group is higher than that in the model group, PBS group and the Huangqi group. The rMSCs + Huangqi group has a higher score than the Huangqi group at each time point. Also, except on day 14, the score for the rMSCs + Huangqi group was higher than the rMSCs group, and the difference was significant. HE staining of the spinal cord paraffin sections showed that the degree of degeneration and necrosis in nerve cells was alleviated in the rMSCs and rMSCs + Huangqi groups, compared with the PBS and Huangqi groups. Edema and infiltration of inflammatory cells were obviously reduced, and the proliferation of glial cells and integrated nerve cells was detected in the rMSCs and rMSCs + Huangqi groups. Moreover, the proliferation of glial cells was more active in the rMSCs + Huangqi group than in the rMSCs group. Using immunohistochemical analysis, both in the rMSCs group and the rMSCs + Huangqi group, BrdU marked rMSCs survived in the spinal cord and accumulated in the gray matter more than in the white matter. GFAP + BrdU and BrdU+NF-M positive cells were detected from day 7 after transplantation. The quantity of BrdU + GFAP positive cells in the rMSCs group and rMSCs + Huangqi group is 6.35±1.14 and 8.75±1.16 per visual field relatively, while BrdU + NF-M positive cells in the rMSCs group and rMSCs + Huangqi group is 1.60±0.75 and 3.30±0.98 per visual field relatively. It indicated that the quantity of BrdU + GFAP and BrdU + NF-M positive cells per visual field observed in the spinal cord paraffin section of the rMSCs + Huangqi group, was significantly higher than that in the rMSCs group (P<0.05). Conclusions These findings indicated that MSCs transplantation has the capability of nerve restoration and recovery after spinal cord injury, and it could be enhanced by Huangqi Injecta. The strategy of combining Huangqi Injecta and MSCs, aimed to promote transplanted cell differentiation and their tissue repair capability, provided a simple yet effective way of optimizing cell transplantation therapy. Disclosures: No relevant conflicts of interest to declare.

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Methylprednisolone inhibits the proliferation of endogenous neural stem cells in nonhuman primates with spinal cord injury

Journal of Neurosurgery Spine ◽

10.3171/2017.12.spine17669 ◽

2018 ◽

Vol 29 (2) ◽

pp. 199-207 ◽

Cited By ~ 7

Author(s):

Jichao Ye ◽

Yi Qin ◽

Yong Tang ◽

Mengjun Ma ◽

Peng Wang ◽

...

Keyword(s):

Spinal Cord ◽

Stem Cells ◽

Spinal Cord Injury ◽

Nonhuman Primates ◽

Therapy Group ◽

Central Canal ◽

Ependymal Cells ◽

Control Group ◽

Cord Injury ◽

Endogenous Neural Stem Cells

OBJECTIVEThe aim of this work was to investigate the effects of methylprednisolone on the proliferation of endogenous neural stem cells (ENSCs) in nonhuman primates with spinal cord injury (SCI).METHODSA total of 14 healthy cynomolgus monkeys (Macaca fascicularis) (4–5 years of age) were randomly divided into 3 groups: the control group (n = 6), SCI group (n = 6), and methylprednisolone therapy group (n = 2). Only laminectomy was performed in the control animals at T-10. SCI was induced in monkeys using Allen’s weight-drop method (50 mm × 50 g) to injure the posterior portion of the spinal cord at T-10. In the methylprednisolone therapy group, monkeys were intravenously infused with methylprednisolone (30 mg/kg) immediately after SCI. All animals were intravenously infused with 5-bromo-2-deoxyuridine (BrdU) (50 mg/kg/day) for 3 days prior to study end point. The small intestine was dissected for immunohistochemical examination. After 3, 7, and 14 days, the spinal cord segments of the control and SCI groups were dissected to prepare frozen and paraffin sections. The proliferation of ENSCs was evaluated using BrdU and nestin immunofluorescence staining.RESULTSHistological examination showed that a larger number of mucosa epithelial cells in the small intestine of all groups were BrdU positive. Nestin-positive ependymal cells are increased around the central canal after SCI. After 3, 7, and 14 days of SCI, BrdU-positive ependymal cells in the SCI group were significantly increased compared with the control group, and the percentage of BrdU-positive cells in the left/right ventral horns and dorsal horn was significantly higher than that of the control group. Seven days after SCI, the percentages of both BrdU-positive ependymal cells around the central canal and BrdU– and nestin–double positive cells in the left/right ventral horns and dorsal horn were significantly lower in the methylprednisolone therapy group than in the SCI group.CONCLUSIONSWhile ENSCs proliferate significantly after SCI in nonhuman primates, methylprednisolone can inhibit the proliferation of ependymal cells after SCI.

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hiPSC-derived NSCs effectively promote the functional recovery of acute spinal cord injury in mice

Stem Cell Research & Therapy ◽

10.1186/s13287-021-02217-9 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Desheng Kong ◽

Baofeng Feng ◽

Asiamah Ernest Amponsah ◽

Jingjing He ◽

Ruiyun Guo ◽

...

Keyword(s):

Spinal Cord ◽

Stem Cells ◽

Spinal Cord Injury ◽

Functional Recovery ◽

Morphological Changes ◽

Embryonic Stem ◽

Control Group ◽

Common Disease ◽

Mice Model ◽

Cord Injury

Abstract Background Spinal cord injury (SCI) is a common disease that results in motor and sensory disorders and even lifelong paralysis. The transplantation of stem cells, such as embryonic stem cells (ESCs), induced pluripotent stem cells (iPSCs), mesenchymal stem cells (MSCs), or subsequently generated stem/progenitor cells, is predicted to be a promising treatment for SCI. In this study, we aimed to investigate effect of human iPSC-derived neural stem cells (hiPSC-NSCs) and umbilical cord-derived MSCs (huMSCs) in a mouse model of acute SCI. Methods Acute SCI mice model were established and were randomly treated as phosphate-buffered saline (PBS) (control group), repaired with 1 × 105 hiPSC-NSCs (NSC group), and 1 × 105 huMSCs (MSC group), respectively, in a total of 54 mice (n = 18 each). Hind limb motor function was evaluated in open-field tests using the Basso Mouse Scale (BMS) at days post-operation (dpo) 1, 3, 5, and 7 after spinal cord injury, and weekly thereafter. Spinal cord and serum samples were harvested at dpo 7, 14, and 21. Haematoxylin-eosin (H&E) staining and Masson staining were used to evaluate the morphological changes and fibrosis area. The differentiation of the transplanted cells in vivo was evaluated with immunohistochemical staining. Results The hiPSC-NSC-treated group presented a significantly smaller glial fibrillary acidic protein (GFAP) positive area than MSC-treated mice at all time points. Additionally, MSC-transplanted mice had a similar GFAP+ area to mice receiving PBS. At dpo 14, the immunostained hiPSC-NSCs were positive for SRY-related high-mobility-group (HMG)-box protein-2 (SOX2). Furthermore, the transplanted hiPSC-NSCs differentiated into GFAP-positive astrocytes and beta-III tubulin-positive neurons, whereas the transplanted huMSCs differentiated into GFAP-positive astrocytes. In addition, hiPSC-NSC transplantation reduced fibrosis formation and the inflammation level. Compared with the control or huMSC transplanted group, the group with transplantation of hiPSC-NSCs exhibited significantly improved behaviours, particularly limb coordination. Conclusions HiPSC-NSCs promote functional recovery in mice with acute SCI by replacing missing neurons and attenuating fibrosis, glial scar formation, and inflammation. Graphical abstract

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Neuroregenerative Effects of Intraspinal Microstimulation (ISMS) Following Spinal Cord Injury (SCI)

Eureka ◽

10.29173/eureka28758 ◽

2021 ◽

Vol 6 (1) ◽

Author(s):

Andy Lee ◽

Mason Schindle ◽

Neil Tyreman ◽

Vivian Mushahwar

Keyword(s):

Spinal Cord ◽

Spinal Cord Injury ◽

Electrical Stimulation ◽

Sham Group ◽

Cellular Damage ◽

Control Group ◽

Intraspinal Microstimulation ◽

Post Surgery ◽

Cord Injury ◽

Control Sham

Background Intraspinal microstimulation (ISMS) is a novel electrical stimulation technique that has demonstrated mobility restoration in animals with spinal cord injury (SCI). This project investigated: 1) the capacity of ISMS to restore functional walking in rats with SCI through 4 weeks of stimulation, and 2) the degree of walking deficit caused by ISMS surgery. Methods Thirteen Sprague Dawley rats were divided into three groups: 1) rats with hemi-section SCI (hSCI) and no implants (control group), 2) rats with hSCI and passive ISMS implants (ISMS sham group), and 3) rats with hSCI and implants with active electrical stimulation (ISMS group). All groups were trained to walk on a horizontal ladder and their performance was quantified pre- and post-surgery. Results We hypothesized that the rats with active ISMS implants would demonstrate the greatest improvement in functional walking compared to both control groups, and that the ISMS sham group would underperform the most. The preoperative functional walking scores of control, sham and ISMS rats were 5.7±0.2, 5.5±0.3 and 5.7±0.1, respectively (7-point scale; mean ± standard error). The post-surgery scores were 3.2±0.9, 2.6±0.6 and 3.3±0.8 for control, sham, and ISMS rats, respectively. Conclusions As the difference between the post-surgery functional walking scores of ISMS and control rats was not statistically significant, this may indicate that four weeks of ISMS stimulation is not enough to cause rehabilitative effects. Additionally, the ISMS sham group demonstrated impaired functional walking compared to the hSCI control group as predicted. Future studies will employ a larger sample size to fully elucidate this trend and utilize thinner microwires to mitigate cellular damage.

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HiPSC-Derived NSCs Effectively Promote The Functional Recovery of Acute Spinal Cord Injury in Mice

10.21203/rs.3.rs-102947/v1 ◽

2020 ◽

Author(s):

Desheng Kong ◽

Baofeng Feng ◽

Asiamah Ernest Amponsah ◽

Jingjing He ◽

Ruiyun Guo ◽

...

Keyword(s):

Spinal Cord ◽

Stem Cells ◽

Spinal Cord Injury ◽

Functional Recovery ◽

Morphological Changes ◽

Embryonic Stem ◽

Control Group ◽

Common Disease ◽

Mice Model ◽

Cord Injury

Abstract Background: Spinal cord injury (SCI) is a common disease that results in motor and sensory disorders and even lifelong paralysis. The transplantation of stem cells, such as embryonic stem cells (ESCs), induced pluripotent stem cells (iPSCs), mesenchymal stem cells (MSCs), or subsequently generated stem/progenitor cells, is predicted to be a promising treatment for SCI. In this study, we aimed to investigate effect of human iPSC-derived neural stem cells (hiPSC-NSCs) and umbilical cord-derived MSCs (huMSCs) in a mouse model of acute SCI. Methods: Acute SCI mice model were established and were randomly treated as PBS (control group), repaired with 1×105 hiPSC-NSCs (NSC group), and 1×105 huMSCs (MSC group), respectively, in a total of 54 mice (n = 18 each). Hind limb motor function was evaluated in open-field tests using the Basso Mouse Scale (BMS) at days post-operation (dpo) 1, 3, 5 and 7 after spinal cord injury, and weekly thereafter. Spinal cord and serum samples were harvested at dpo 7, 14 and 21. HE staining and Masson staining were used to evaluate the morphological changes and fibrosis area. The differentiation of the transplanted cells in vivo was evaluated with immunohistochemical staining. Results: The hiPSC-NSC-treated group presented a significantly smaller GFAP+ area than MSC-treated mice at all time points. Additionally, MSC-transplanted mice had a similar GFAP+ area to mice receiving PBS. At dpo14, the immunostained hiPSC-NSCs were positive for SOX2. Furthermore, the transplanted hiPSC-NSCs differentiated into glial fibrillary acidic protein (GFAP)-positive astrocytes and beta-III tubulin-positive neurons, whereas the transplanted huMSCs differentiated into GFAP-positive astrocytes. In addition, hiPSC-NSC transplantation reduced fibrosis formation and the inflammation level. Compared with the control or huMSC transplanted group, the group with transplantation of hiPSC-NSCs exhibited significantly improved behaviours, particularly limb coordination. Conclusions: HiPSC-NSCs promote functional recovery in mice with acute SCI by replacing missing neurons and attenuating fibrosis, glial scar formation, and inflammation.

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Study on Application of Bone Marrow Mesenchymal Stem Cells Transplantation in Restorative Treatment of Spinal Cord Injury in Mice

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2020.2404 ◽

2020 ◽

Vol 10 (8) ◽

pp. 1122-1127

Author(s):

Yang Sun ◽

Zhongjing Jiang ◽

Hao Tang ◽

Jiang Xie

Keyword(s):

Spinal Cord ◽

Stem Cells ◽

Bone Marrow ◽

Spinal Cord Injury ◽

Mesenchymal Stem Cells ◽

Nerve Fibers ◽

Control Group ◽

Mouse Spinal Cord ◽

Cord Injury ◽

Marrow Mesenchymal Stem Cells

The spinal cord injury (SCI) refers to different degrees of injuries in the structure or function of spinal cord caused by different factors. The prevalence rate of SCI in the population under 40 years reaches 80%. SCI causes certain injury to both physiology and psychology of patients. An important factor leading to SCI is the rupture of nerve fibers. Bone marrow mesenchymal stem cells (BMMSCs) have the functions of inducing and supporting hematopoietic stem cells in bone marrow. In this study, SCI mouse model was established to assess the effect of BM-MSCs on SCI. A total of 30 SCI mouse model were established and assigned into transplantation group (15 mice) and control group (15 mice) according to random number table method. The mice in transplantation group were treated with BM-MSCs transplantation at SCI site, while mice in control group were treated with normal saline at SCI site. The bone marrow pathological changes were measured by HE staining and neural cells were assessed by Pischingert's methylene blue staining along with measuring SRY level by immunohistochemistry. The motor abilities of mice in transplantation group at the 2nd, 4th, 6th and 8th weeks were significantly higher than mice in control group (P < 0 05). A few vacuoles appeared in mice in transplantation group. The number of cells in mouse spinal cord tissues in transplantation group was increased significantly over time, but a large number of vacuoles appeared in mouse spinal cord tissues in control group with necrosis of a vast amount of nerve fibers (P < 0 05). The number and volume of Nissl bodies in mice in transplantation group was increased significantly at 2 weeks after treatment and degeneration status of nerve cells in transplantation group was significantly better than control group (P < 0 05). The SRY genes were expressed in transplantation group for a long term but not in control group (P < 0 05). The number of adherent cells increased significantly in transplantation group at 48 hours after treatment. BMMSCs transplantation can effectively promotes the recovery of SCI mice, indicating that it is worthy of clinical promotion and application.

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Stem Cell Transplantation: A Promising Therapy for Spinal Cord Injury

Current Stem Cell Research & Therapy ◽

10.2174/1574888x14666190823144424 ◽

2020 ◽

Vol 15 (4) ◽

pp. 321-331 ◽

Cited By ~ 2

Author(s):

Zhe Gong ◽

Kaishun Xia ◽

Ankai Xu ◽

Chao Yu ◽

Chenggui Wang ◽

...

Keyword(s):

Spinal Cord ◽

Stem Cells ◽

Spinal Cord Injury ◽

Stem Cell ◽

Stem Cell Transplantation ◽

Cell Transplantation ◽

Therapeutic Potential ◽

Embryonic Stem ◽

Current Status ◽

Cord Injury

Spinal Cord Injury (SCI) causes irreversible functional loss of the affected population. The incidence of SCI keeps increasing, resulting in huge burden on the society. The pathogenesis of SCI involves neuron death and exotic reaction, which could impede neuron regeneration. In clinic, the limited regenerative capacity of endogenous cells after SCI is a major problem. Recent studies have demonstrated that a variety of stem cells such as induced Pluripotent Stem Cells (iPSCs), Embryonic Stem Cells (ESCs), Mesenchymal Stem Cells (MSCs) and Neural Progenitor Cells (NPCs) /Neural Stem Cells (NSCs) have therapeutic potential for SCI. However, the efficacy and safety of these stem cellbased therapy for SCI remain controversial. In this review, we introduce the pathogenesis of SCI, summarize the current status of the application of these stem cells in SCI repair, and discuss possible mechanisms responsible for functional recovery of SCI after stem cell transplantation. Finally, we highlight several areas for further exploitation of stem cells as a promising regenerative therapy of SCI.

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Exercise Ameliorates Spinal Cord Injury by Changing DNA Methylation

Cells ◽

10.3390/cells10010143 ◽

2021 ◽

Vol 10 (1) ◽

pp. 143

Author(s):

Ganchimeg Davaa ◽

Jin Young Hong ◽

Tae Uk Kim ◽

Seong Jae Lee ◽

Seo Young Kim ◽

...

Keyword(s):

Spinal Cord ◽

Spinal Cord Injury ◽

Motor Cortex ◽

Functional Recovery ◽

Treadmill Exercise ◽

Exercise Group ◽

Control Group ◽

Epigenetic Changes ◽

Cord Injury ◽

The Brain

Exercise training is a traditional method to maximize remaining function in patients with spinal cord injury (SCI), but the exact mechanism by which exercise promotes recovery after SCI has not been identified; whether exercise truly has a beneficial effect on SCI also remains unclear. Previously, we showed that epigenetic changes in the brain motor cortex occur after SCI and that a treatment leading to epigenetic modulation effectively promotes functional recovery after SCI. We aimed to determine how exercise induces functional improvement in rats subjected to SCI and whether epigenetic changes are engaged in the effects of exercise. A spinal cord contusion model was established in rats, which were then subjected to treadmill exercise for 12 weeks. We found that the size of the lesion cavity and the number of macrophages were decreased more in the exercise group than in the control group after 12 weeks of injury. Immunofluorescence and DNA dot blot analysis revealed that levels of 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) in the brain motor cortex were increased after exercise. Accordingly, the expression of ten-eleven translocation (Tet) family members (Tet1, Tet2, and Tet3) in the brain motor cortex also elevated. However, no macrophage polarization was induced by exercise. Locomotor function, including Basso, Beattie, and Bresnahan (BBB) and ladder scores, also improved in the exercise group compared to the control group. We concluded that treadmill exercise facilitates functional recovery in rats with SCI, and mechanistically epigenetic changes in the brain motor cortex may contribute to exercise-induced improvements.

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Bone marrow mesenchymal stem cells-derived exosomes reduce apoptosis and inflammatory response during spinal cord injury by inhibiting the TLR4/MyD88/NF-κB signaling pathway

Human & Experimental Toxicology ◽

10.1177/09603271211003311 ◽

2021 ◽

pp. 096032712110033

Author(s):

Liying Fan ◽

Jun Dong ◽

Xijing He ◽

Chun Zhang ◽

Ting Zhang

Keyword(s):

Spinal Cord ◽

Stem Cells ◽

Bone Marrow ◽

Spinal Cord Injury ◽

Mesenchymal Stem Cells ◽

Motor Function ◽

Inflammatory Factors ◽

Cord Injury ◽

Marrow Mesenchymal Stem Cells ◽

Apoptosis And Inflammation

Spinal cord injury (SCI) is one of the most common destructive injuries, which may lead to permanent neurological dysfunction. Currently, transplantation of bone marrow mesenchymal stem cells (BMSCs) in experimental models of SCI shows promise as effective therapies. BMSCs secrete various factors that can regulate the microenvironment, which is called paracrine effect. Among these paracrine substances, exosomes are considered to be the most valuable therapeutic factors. Our study found that BMSCs-derived exosomes therapy attenuated cell apoptosis and inflammation response in the injured spinal cord tissues. In in vitro studies, BMSCs-derived exosomes significantly inhibited lipopolysaccharide (LPS)-induced PC12 cell apoptosis, reduced the secretion of pro-inflammatory factors including tumor necrosis factor (TNF)-α and IL (interleukin)-1β and promoted the secretion of anti-inflammatory factors including IL-10 and IL-4. Moreover, we found that LPS-induced protein expression of toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88) and nuclear transcription factor-κB (NF-κB) was significantly downregulated after treatment with BMSCs-derived exosomes. In in vivo studies, we found that hindlimb motor function was significantly improved in SCI rats with systemic administration of BMSCs-derived exosomes. We also observed that the expression of pro-apoptotic proteins and pro-inflammatory factors was significantly decreased, while the expression of anti-apoptotic proteins and anti-inflammatory factors were upregulated in SCI rats after exosome treatment. In conclusion, BMSCs-derived exosomes can inhibit apoptosis and inflammation response induced by injury and promote motor function recovery by inhibiting the TLR4/MyD88/NF-κB signaling pathway, which suggests that BMSCs-derived exosomes are expected to become a new therapeutic strategy for SCI.

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