A combination therapy for slowing microglia/macrophage-mediated inflammatory response after spinal cord injury
A technology for microglia and spinal cord injury, applied in the field of neuromedicine, can solve problems such as hindering the repair of spinal cord injury, and poor relieving effect, achieving the effects of promoting endogenous nerve regeneration and functional recovery, promoting clearance, and promoting repair.
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Embodiment 1
[0045] Example 1 Preparation and Performance Analysis of Gelatin Hydrogel Grafting Materials
[0046] The present embodiment provides the preparation method of the gelatin hydrogel that is used for spinal cord injury site transplantation, specifically as follows:
[0047] (1) Prepare methacrylated gelatin (Methacrylated Gelalin, mGL) in aqueous solution by grafting gelatin with methacrylic anhydride; make mGL into an 8% solution under sterile conditions, and titrate to pH7 with HCl or NaOH .0.
[0048] (2) Adding the photoinitiator to the mGL solution obtained in step (1) to make the final concentration 0.08% (w / v) to obtain an aqueous gelatin solution containing the photoinitiator.
[0049] (3) The obtained mixture solution is irradiated with a visible light source or an ultraviolet light source for 180 seconds to completely solidify to obtain a gelatin hydrogel.
[0050] After the gelatin hydrogel prepared above was freeze-dried, it was detected by scanning electron micros...
Embodiment 2
[0051] Example 2 PLX3397 treatment on the elimination of microglia in uninjured spinal cord and safety evaluation
[0052] 1. Clearance and repopulation of microglial cells in the spinal cord after PLX3397 treatment
[0053] Ten healthy mice were selected and randomly divided into two groups (PLX3397 treatment group and control group). The PLX3397 treatment group was fed with a feed containing 290 mg / kg PLX3397 (free feeding), and the control group was fed with a feed without PLX3397. On the 14th day, the number of microglial cells was detected by immunofluorescence, and the results were as follows: figure 2 showed that about 95% of the microglia in the spinal cord could be eliminated ( figure 2 A and C). Moreover, the number of microglial cells in the PLX3397 treatment group could return to the initial level after 14 days of stopping administration ( figure 2 B and D). The results showed that the treatment of CSF1R inhibitor PLX3397 could effectively clear the microgli...
Embodiment 3
[0060] Example 3 Combination therapy can effectively reduce the inflammatory response mediated by microglia / macrophages after total transected spinal cord injury
[0061] The results of Example 2 above show that PLX3397 can safely and effectively regulate microglia in the spinal cord in healthy mice. Activation of microglia / macrophage-mediated inflammatory responses following injury.
[0062] 1. Combination therapy can specifically regulate the clearance and repopulation of microglia after spinal cord injury
[0063] 5 mice were selected for each group, and after being anesthetized with pentobarbital sodium, 1 mm of tissue was completely removed from the T7-T8 segment of the spinal cord for total transection defect spinal cord injury. The photoinitiator-containing gelatin aqueous solution prepared in step (2) of Example 1 was cross-linked for 180 seconds under light conditions to form a gelatin hydrogel. After the mice woke up and recovered, the experimental group and the co...
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