中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (14): 3652-3662.doi: 10.12307/2026.077

• 水凝胶材料Hydrogel materials • 上一篇    下一篇

负载无细胞脂肪提取物的导电水凝胶修复大鼠脊髓损伤

杨  标1，2，吴中桓1，2，姜福贵1，2，何成龙1，2，李廷栋1，2   

  1. 1黔东南苗族侗族自治州人民医院骨科，贵州省凯里市   556000；2 黔东南州临床医学研究中心，贵州省凯里市   556000
  • 收稿日期:2025-03-06 接受日期:2025-05-17 出版日期:2026-05-18 发布日期:2025-09-12
  • 通讯作者: 杨标，博士，副主任医师，黔东南苗族侗族自治州人民医院骨科，贵州省凯里市 556000；黔东南州临床医学研究中心，贵州省凯里市 556000
  • 作者简介:杨标，男，1985年生，贵州省凯里市人，侗族，博士，副主任医师，主要从事脊髓损伤的再生修复研究。
  • 基金资助:
    黔东南州基础研究计划(黔东南科合基础[2023]21号)，项目负责人：杨标

Conductive hydrogel with cell-free fat extract repairs spinal cord injuries in rats

Yang Biao1, 2, Wu Zhonghuan1, 2, Jiang Fugui1, 2, He Chenglong1, 2, Li Tingdong1, 2   

  1. 1Department of Orthopedics, People’s Hospital of Qiandongnan Miao and Dong Autonomous Prefecture, Kaili 556000, Guizhou Province, China; 2Qiandongnan Clinical Medical Research Center, Kaili 556000, Guizhou Province, China
  • Received:2025-03-06 Accepted:2025-05-17 Online:2026-05-18 Published:2025-09-12
  • Contact: Yang Biao, MD, Associate chief physician, Department of Orthopedics, People’s Hospital of Qiandongnan Miao and Dong Autonomous Prefecture, Kaili 556000, Guizhou Province, China; Qiandongnan Clinical Medical Research Center, Kaili 556000, Guizhou Province, China
  • About author:Yang Biao, MD, Associate chief physician, Department of Orthopedics, People’s Hospital of Qiandongnan Miao and Dong Autonomous Prefecture, Kaili 556000, Guizhou Province, China; Qiandongnan Clinical Medical Research Center, Kaili 556000, Guizhou Province, China
  • Supported by:
    Qiandongnan Prefecture Basic Research Program, Qiandongnan Kehe Foundation [2023] No. 21 (to YB)

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摘要：

文题释义：
功能化导电水凝胶：是由导电材料和交联水凝胶聚合物网络组成，具有类组织的力学性能和与组织的电相互作用能力。在损伤区植入功能化导电水凝胶可以通过重塑脊髓损伤的微环境，促进轴突再生和神经元回路的生成。
无细胞脂肪提取物：是一种从人皮下脂肪组织中提取的非细胞液，不含细胞成分和脂质残留物，无免疫原性，生物安全性好。无细胞脂肪提取物的成分类似于干细胞旁分泌因子，这些因子可促进新生血管形成并具有抗炎作用，还可促进细胞增殖和胶原蛋白分泌。

背景：导电水凝胶有助于脊髓内的内源性神经再生、抑制胶质纤维化形成并建立神经桥网络，可促进脊髓半切损伤模型大鼠运动功能的恢复，但其减轻脊髓损伤后炎症反应的能力有限。无细胞脂肪提取物具有免疫调节和促进组织再生作用。
目的：探讨负载无细胞脂肪提取物超分子导电水凝胶在大鼠脊髓损伤修复中的作用。
方法：①从SD大鼠腹股沟脂肪组织中提取无细胞脂肪提取物。分别制备琼脂/明胶水凝胶与琼脂/明胶/聚吡咯水凝胶，表征水凝胶的微观形貌、流变性、溶胀率与导电性。制备负载无细胞脂肪提取物的琼脂/明胶/聚吡咯水凝胶，检测水凝胶的体外释药性能。②取80只SD大鼠建立脊髓损伤半切模型，随机分4组干预：模型组(n=20)不进行任何干预，普通水凝胶组(n=20)、导电水凝胶组(n=20)、载药导电水凝胶组(n=20)损伤脊髓部位分别植入琼脂/明胶水凝胶、琼脂/明胶/聚吡咯水凝胶及负载无细胞脂肪提取物的琼脂/明胶/聚吡咯水凝胶。术后第42天，利用BBB评分和斜板实验评估大鼠右侧后肢运动功能情况，苏木精-伊红染色观察大鼠脊髓损伤组织形态，免疫荧光检测脊髓损伤中心CD86、CD206、微管相关蛋白2、微管蛋白βⅢ、硫酸软骨素56、神经丝蛋白200及髓鞘碱性蛋白表达，牢固蓝染色与透射电子显微镜评估轴突髓鞘再生程度；术后14 d，ELISA法检测损伤脊髓组织中肿瘤坏死因子α、白细胞介素1β、白细胞介素10和白细胞介素6水平。
结果与结论：①琼脂/明胶水凝胶与琼脂/明胶/聚吡咯水凝胶呈相互连接的多孔结构，具有较高的孔隙率与流变性，琼脂/明胶/聚吡咯水凝胶的孔径、溶胀率低于琼脂/明胶水凝胶(P < 0.05)，导电率高于琼脂/明胶水凝胶(P < 0.05)。浸泡于PBS中后，无细胞脂肪提取物从琼脂/明胶/聚吡咯水凝胶表现出缓慢且持续的释放，5 d的累积释放率约为31%。②相较于其他3组，载药导电水凝胶组大鼠后肢运动功能得到明显改善，脊髓损伤区域囊腔面积显著减少，脊髓组织中肿瘤坏死因子α、白细胞介素1β、白细胞介素6水平下降及白细胞介素10水平升高，CD86与硫酸软骨素56表达显著降低，CD206、微管相关蛋白2、微管蛋白βⅢ、神经丝蛋白200及髓鞘碱性蛋白表达显著升高，髓鞘再生明显。结果表明载药导电水凝胶降低了大鼠损伤脊髓组织中小胶质细胞/巨噬细胞的活性和促炎因子的表达、促进内源性神经干细胞分化为神经元，最终促进轴突再生、再生轴突髓鞘形成及神经功能恢复。
https://orcid.org/0009-0007-1729-0437(杨标) 

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料；口腔生物材料；纳米材料；缓释材料；材料相容性；组织工程

关键词: 脊髓损伤, 导电聚合物, 水凝胶, 电活性生物材料, 无细胞脂肪提取物, 神经再生, 工程化脊髓材料

Abstract: BACKGROUND: Conductive hydrogels can help endogenous nerve regeneration in the spinal cord, inhibit the formation of glial fibrosis and establish a neural bridge network, and promote the recovery of motor function in rats with hemisection injury model, but their ability to reduce inflammatory response after spinal cord injury is limited. Cell-free fat extract has immunomodulatory and tissue regeneration promoting effects. 
OBJECTIVE: To explore the role of supramolecular conductive hydrogel loaded with cell-free fat extract in the repair of spinal cord injury in rats.
METHODS: (1) Cell-free fat extract was extracted from the inguinal adipose tissue of SD rats. Agar/gelatin hydrogel and agar/gelatin/polypyrrole hydrogel were prepared respectively, and the micromorphology, rheology, swelling rate, and conductivity of the hydrogel were characterized. Agar/gelatin/polypyrrole hydrogel loaded with cell-free fat extract was prepared, and the in vitro drug release performance of the hydrogel was tested. (2) A total of 80 SD rats were used to establish a hemisection model of spinal cord injury and randomly divided into four intervention groups: the model group (n=20) received no intervention; the ordinary hydrogel group (n=20), the conductive hydrogel group (n=20), and the drug-loaded conductive hydrogel group (n=20) were implanted with agar/gelatin hydrogel, agar/gelatin/polypyrrole hydrogel, and agar/gelatin/polypyrrole hydrogel loaded with cell-free fat extract at the injured spinal cord, respectively. On day 42 after surgery, the motor function of the right hind limb of the rats was evaluated by BBB score and inclined board test. The morphology of the spinal cord injury tissue of the rats was observed by hematoxylin-eosin staining. The expression levels of CD86, CD206, microtubule-associated protein 2, tubulin βIII, chondroitin sulfate 56, neurofilament protein 200, and myelin basic protein in the spinal cord injury center were detected by immunofluorescence. The degree of axonal myelination was evaluated by fast blue staining and transmission electron microscopy. On day 14 after surgery, the levels of tumor necrosis factor α, interleukin 1β, interleukin 10, and interleukin 6 in the injured spinal cord tissue were detected by ELISA.
RESULTS AND CONCLUSION: (1) Agar/gelatin hydrogel and agar/gelatin/polypyrrole hydrogel presented interconnected porous structures with high porosity and rheological properties. The pore size and swelling rate of agar/gelatin/polypyrrole hydrogel were lower than those of agar/gelatin hydrogel (P < 0.05), and the conductivity was higher than that of agar/gelatin hydrogel (P < 0.05). After soaking in PBS, the cell-free fat extract showed slow and sustained release from the agar/gelatin/polypyrrole hydrogel, with a cumulative release rate of about 31% in 5 days. (2) Compared with the other three groups, the motor function of the hind limbs of rats in the drug-loaded conductive hydrogel group was significantly improved, the area of the cyst in the spinal cord injury area was significantly reduced, the levels of tumor necrosis factor α, interleukin 1β, and interleukin 6 in the spinal cord tissue decreased, and the level of interleukin 10 increased. The expression of CD86 and chondroitin sulfate 56 was significantly reduced, and the expression of CD206, microtubule-associated protein 2, tubulin βIII, neurofilament protein 200, and myelin basic protein was significantly increased, and myelin regeneration was obvious. The results showed that the drug-loaded conductive hydrogel reduced the activity of microglia/macrophages and the expression of proinflammatory factors in the injured spinal cord tissue of rats, promoted the differentiation of endogenous neural stem cells into neurons, and ultimately promoted axon regeneration, myelination of regenerated axons, and recovery of neural function.

Key words: spinal cord injury, conductive polymer, hydrogel, electroactive biomaterial, cell-free fat extract, neural regeneration, engineered spinal cord material

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