基于生物3D打印技术的丹酚酸B-海藻酸钠-明胶皮肤支架治疗糖尿病创面的实验研究
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1.南京医科大学附属常州二院;2.大连医科大学研究生院;3.南京医科大学附属常州第二人民医院;4.淮安市淮安医院(淮安市肿瘤医院)

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江苏省中医药科技发展计划面上项目(MS2021056);常州市应用基础研究项目(CJ20220258)


Experimental study on the treatment of diabetic wound with salvianolic acid B-sodium alginate-gelatin skin scaffolds produced by biological 3D printing technology
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The Affiliated Changzhou No. 2 People’s Hospital with Nanjing Medical University

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    摘要:

    目的:探讨采用生物3D打印技术制备的丹酚酸B(SAB)-海藻酸钠-明胶皮肤支架治疗糖尿病创面的体内生物相容性及疗效。 方法:按照SAB占海藻酸钠与明胶总重的百分比(wt%)制成0、0.5、1.0、1.5 wt%的生物墨水,采用生物3D打印技术制备不同规格的皮肤支架。扫描电镜观察微观结构,高效液相色谱测定SAB体外累积释放药物浓度。皮肤支架用于治疗2型糖尿病大鼠创面,于第7d、14d观察创面愈合、渗出情况,同时取创面组织进行HE、Masson、活性氧(ROS)染色,并检测超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-PX)、过氧化氢酶(CAT)及丙二醛(MDA)水平。 结果:五组皮肤支架均呈网孔状立体结构,孔隙分布均匀。药物累积释放量随时间延长逐渐增加。大体观察:各组创面均愈合良好,1.0%SAB组创面修复优于其他各组。第7d及14d时,1.0%SAB组ROS水平均低于其他各组(P<0.05);第7d时,1.0%SAB组SOD、GSH-PX及CAT水平均高于其他各组(P<0.005),MDA水平低于空白对照组、凡士林纱布组及0%SAB组;第14d时,1.0%SAB组SOD、GSH-PX及CAT水平高于空白对照组、凡士林纱布组及0%SAB组(P<0.005),MDA水平低于空白对照组、凡士林纱布组(P<0.05)。HE染色:各组创面皮肤均修复较好,1.0%SAB组新生组织较多。Masson染色:第7d及14d时,0.5、1.0、1.5%SAB组创面胶原蛋白沉积多于其他各组(P<0.05)。 结论:1.0%SAB-海藻酸钠-明胶皮肤支架不但可以抑制ROS的产生,并上调各种抗氧化酶的表达,从而抑制创面组织内氧化应激反应,还可以促进胶原的沉积,最终促进糖尿病溃疡创面的修复。

    Abstract:

    Objective: To investigate the in vivo biocompatibility and efficacy of salvianolic acid B (SAB) -sodium alginate-gelatin skin scaffold prepared by 3D bioprinting technology in the treatment of diabetic wounds. Methods: The 0, 0.5, 1.0 and 1.5 wt% of bioinks were prepared according to the percentage of SAB in the total weight of sodium alginate and gelatin (wt%). Skin scaffolds with different type were produced by biological 3D printing technology. The microstructure was observed by scanning electron microscopy, and the concentration of SAB in vitro was determined by High-performance liquid chromatography. The skin scaffold was used in the treatment of ulcer of rats with type 2 diabetes milltus, and the healing and exudation of the ulcer were observed and photo-recorded on the 7th and 14th day. Meanwhile, the wound tissue was taken for HE, Masson, reactive oxygen species (ROS) staining, and the levels of superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), catalase (CAT), and malondialdehyde (MDA) were detected. Results: All the skin scaffolds showed a mesh-like three-dimensional structure with uniform pore distribution. The cumulative release of the drug increases gradually over time. General observations: wounds healed well in each group, 1% groups healing better than other groups. On days 7 and 14, ROS level in 1.0%SAB group was lower than that in other groups (P<0.05). On day 7, the levels of SOD, GSH-PX and CAT in 1.0%SAB group were higher than those in other groups (P<0.005), MDA level was lower than that of the control group, vaseline gauze group and 0%SAB group. On day 14, the levels of SOD, GSH-PX and CAT in 1.0%SAB group were higher than those in control group, vasolin gauze group and 0%SAB group (P<0.005), MDA level was lower than that in the control group and vaseline gauze group (P<0.05). HE staining: the wounds was well repaired of each group, and more new tissue was formed in the 1.0%SAB group. Masson staining: on days 7 and 14, collagen deposition in 0.5, 1.0, 1.5% groups was more than that in other groups (P<0.05). Conclusion: The 1.0% SAB-alginate-gelatin skin scaffold can not only inhibit the production of ROS and up-regulate the expression of various antioxidant enzymes, thereby inhibiting oxidative stress response in wound tissue, but also promote the deposition of collagen, and ultimately promote the repair of diabetic ulcer wound.

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  • 收稿日期:2023-10-10
  • 最后修改日期:2023-12-12
  • 录用日期:2024-02-27
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