Chinese Journal of Tissue Engineering Research ›› 2024, Vol. 28 ›› Issue (3): 380-386.doi: 10.12307/2023.883
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Cao Sheng, Kong Lingwei, Xu Kun, Sun Zhijie
Received:
2022-11-04
Accepted:
2022-12-09
Online:
2024-01-28
Published:
2023-07-10
Contact:
Xu Kun, Master, Associate chief physician, Affiliated Hospital of Chengde Medical College, Chengde 067000, Hebei Province, China
About author:
Cao Sheng, Master, Attending physician, Affiliated Hospital of Chengde Medical College, Chengde 067000, Hebei Province, China
Supported by:
CLC Number:
Cao Sheng, Kong Lingwei, Xu Kun, Sun Zhijie. Effect of gelatin methacryloyl hydrogel loaded with salvianolic acid B on intervertebral disc degeneration[J]. Chinese Journal of Tissue Engineering Research, 2024, 28(3): 380-386.
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2.4 体外细胞实验结果 2.4.1 各组细胞增殖实验 CCK-8检测结果显示,5组细胞随着培养时间的增加持续增殖,见图4。培养第1天时,与A组比较,B组细胞增殖吸光度值显著降低(P < 0.05);与B组比较,C、D、E组细胞增殖吸光度值无明显变化(P > 0.05)。培养第3天时,与A组比较,B组细胞增殖吸光度值显著降低(P < 0.05);与B组比较,C组细胞增殖吸光度值无明显变化(P > 0.05),D组细胞增殖吸光度值显著升高(P < 0.05);与D组比较,E组细胞增殖吸光度值显著升高(P < 0.05)。培养第7天时,与A组比较,B组细胞增殖吸光度值显著降低(P < 0.05);与B组比较,C组细胞增殖吸光度值无明显变化(P > 0.05),D组细胞增殖吸光度值显著升高(P < 0.05);与D组比较,E组细胞增殖吸光度值显著升高(P < 0.05)。"
与A组比较,B组Ⅱ型胶原、蛋白聚糖的mRNA表达量明显减少(P < 0.05),基质金属蛋白酶3、基质金属蛋白酶13、ADAMTS4、ADAMTS5的mRNA表达量明显增加(P < 0.05)。与B组比较,C组各基因的mRNA表达量无显著变化(P > 0.05),D、E组Ⅱ型胶原、蛋白聚糖的mRNA表达量明显增加(P < 0.05),基质金属蛋白酶3、基质金属蛋白酶13、ADAMTS4、ADAMTS5的mRNA表达量明显减少(P < 0.05)。与D组比较,E组Ⅱ型胶原、蛋白聚糖的mRNA表达量明显增加(P < 0.05),基质金属蛋白酶3、基质金属蛋白酶13、ADAMTS4、ADAMTS5的mRNA表达量明显减少(P < 0.05)。 2.4.5 Western blot检测各组细胞TLR4/核因子κB 信号通路蛋白的表达 见图7。"
与A组比较,B组高迁移率族蛋白、TLR4、Ik Bα和核因子κB P65的蛋白表达量明显增加(P < 0.05)。与B组比较,C组高迁移率族蛋白、TLR4、Ik Bα和核因子κB P65的蛋白表达量无显著变化(P > 0.05),D、E组高迁移率族蛋白、TLR4、Ik Bα和核因子κB P65的蛋白表达量明显减少(P < 0.05)。与D组比较,E组高迁移率族蛋白、TLR4、Ik Bα和核因子κB P65的蛋白表达量明显减少(P < 0.05)。 2.5 动物体内实验结果 2.5.1 各组大鼠椎间盘X射线片检测 见图8A、B。 术后0周时,针刺组、针刺+丹酚酸组、针刺+水凝胶组、针刺+载药水凝胶组椎间盘高度指数均低于正常组(P < 0.05);针刺组、针刺+丹酚酸组、针刺+水凝胶组、针刺+载药水凝胶组椎间盘高度指数比较差异无显著性意义(P > 0.05)。术后4周时,针刺组、针刺+丹酚酸组、针刺+水凝胶组、针刺+载药水凝胶组椎间盘高度指数均低于正常组(P < 0.05);与针刺组比较,针刺+丹酚酸组、针刺+水凝胶组、针刺+载药水凝胶组椎间盘高度指数升高(P < 0.05);与针刺+丹酚酸组、针刺+水凝胶组比较,针刺+载药水凝胶组椎间盘高度指数升高(P < 0.05)。 2.5.2 各组大鼠椎间盘MRI检查 见图8C、D。 术后0周时,针刺组、针刺+丹酚酸组、针刺+水凝胶组、针刺+载药水凝胶组MRI指数均低于正常组(P < 0.05);针刺组、针刺+丹酚酸组、针刺+水凝胶组、针刺+载药水凝胶组MRI指数比较差异无显著性意义(P > 0.05)。术后4周时,针刺组、针刺+丹酚酸组、针刺+水凝胶组、针刺+载药水凝胶组MRI指数均低于正常组(P < 0.05);与针刺组比较,针刺+丹酚酸组、针刺+水凝胶组、针刺+载药水凝胶组MRI指数升高(P < 0.05);与针刺+丹酚酸组、针刺+水凝胶组比较,针刺+载药水凝胶组MRI指数升高(P < 0.05)。"
苏木精-伊红染色显示,正常组中细胞和细胞外基质组织良好;针刺组髓核结构不完整,纤维环层片状结构混乱,退变位点被纤维软骨组织和不同细胞形态的细胞所占据,损伤部位存在大量的瘢痕与纤维结缔组织,椎间盘结构模糊不清;其余3组损伤部位存在瘢痕和纤维结缔组织,部分椎间盘结构模糊不清,其中针刺+载药水凝胶组细胞和细胞外基质结构较针刺+丹酚酸组、针刺+水凝胶组清晰、规则。番红O染色显示,正常组可见完整的髓核组织,含有丰富的胶原组织;针刺组髓核结构不完整,仅见少量的胶原组织;其余3组髓核组织结构破坏程度均轻于针刺组,胶原含量增加,其中以针刺+载药水凝胶组髓核组织结构更细清晰、规则且胶原含量最多。 椎间盘组织学评分结果显示,正常组、针刺组、针刺+丹酚酸组、针刺+水凝胶组、针刺+载药水凝胶组目标椎间盘组织学评分依次为4.39±0.12,13.09±0.13,9.38±0.12,10.54±0.18,7.03±0.22。针刺组、针刺+丹酚酸组、针刺+水凝胶组、针刺+载药水凝胶组评分高于正常组(P < 0.05),针刺+丹酚酸组、针刺+水凝胶组、针刺+载药水凝胶组评分低于针刺组(P < 0.05),针刺+载药水凝胶组评分低于针刺+丹酚酸组、针刺+水凝胶组(P < 0.05)。 2.6 水凝胶的生物相容性 由体外细胞实验与动物体内实验可知,负载丹酚酸B的甲基丙烯酰化明胶水凝胶具有良好的生物相容性。"
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