Chinese Journal of Tissue Engineering Research ›› 2024, Vol. 28 ›› Issue (17): 2667-2674.doi: 10.12307/2024.480
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Liu Ziyang, Lao An, Xu Chenci, AiRi Shin, Wu Jiaqing, Liu Jiaqiang
Received:
2023-08-02
Accepted:
2023-09-18
Online:
2024-06-18
Published:
2023-12-14
Contact:
Liu Jiaqiang, Chief physician, Professor, Department of Oral & Cranio-Maxillofacial Surgery, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine; College of Stomatology, Shanghai Jiao Tong University; National Center for Stomatology; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology, Shanghai Research Institute of Stomatology, Shanghai 200011, China
About author:
Liu Ziyang, Master candidate, Department of Oral & Cranio-Maxillofacial Surgery, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine; College of Stomatology, Shanghai Jiao Tong University; National Center for Stomatology; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology, Shanghai Research Institute of Stomatology, Shanghai 200011, China
Lao An, Doctoral candidate, Department of Oral & Cranio-Maxillofacial Surgery, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine; College of Stomatology, Shanghai Jiao Tong University; National Center for Stomatology; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology, Shanghai Research Institute of Stomatology, Shanghai 200011, China
Supported by:
CLC Number:
Liu Ziyang, Lao An, Xu Chenci, AiRi Shin, Wu Jiaqing, Liu Jiaqiang. Polycaprolactone-polydopamine-AOPDM1 scaffold promotes bone formation in a high-glucose environment[J]. Chinese Journal of Tissue Engineering Research, 2024, 28(17): 2667-2674.
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2.1 支架表征结果 2.1.1 支架形貌 扫描电镜显示3组支架均为直径较均匀、表面连续的纳米级细丝,无规则互相交织形成随机定向的多孔结构,见图1,该多孔结构有利于细胞的浸润、营养物质及气体的输送,并且在结构上仿生了细胞外基质[33-34]。与PCL支架相比,PCL-PDA与PCL-PDA-AOPDM1支架纤维直径逐渐增大,表面也更加粗糙,可见颗粒状沉积,沉积的颗粒分别为PDA和PDA-AOPDM1。图2表面元素分析显示,PCL支架表面的元素主要为C和O;PCL-PDA支架表面的主要元素仍为C和O,出现少量N元素;PCL-PDA-AOPDM1支架表面的元素中出现较多的N,提示多肽成功负载于电纺膜上。"
2.2 AOPDM1对骨髓间充质干细胞增殖与成骨分化的影响 2.2.1 小鼠骨髓间充质干细胞形态观察 图6,7显示小鼠骨髓间充质干细胞的形态,在原代培养初期有一些杂细胞,贴壁细胞于1周左右长满瓶底,随着换液与细胞传代,杂细胞数目逐渐减少,细胞形态逐渐均一化,细胞多为梭形、纺锤形。 2.2.2 AOPDM1对骨髓间充质干细胞增殖的影响 图8显示正常环境与高糖环境下不同浓度AOPDM1对骨髓间充质干细胞增殖的影响。培养第1天,各组细胞增殖无明显差异(P > 0.05)。培养第3天,高糖环境下的骨髓间充质干细胞增殖与正常对照组比较差异无显著性意义(P > 0.05);高糖环境下,6.25,12.5,25,50,100 μmol/L AOPDM1均可促进骨髓间充质干细胞的增殖(P < 0.05,P < 0.01)。培养第7天,高糖环境下0 μmol/L AOPDM1组骨髓间充质干细胞的增殖低于正常对照组(P < 0.001);高糖环境下,12.5,25,50,100 μmol/L的AOPDM1可促进骨髓间充质干细胞的增殖(P < 0.05,P < 0.001)。 2.2.3 AOPDM1对骨髓间充质干细胞碱性磷酸酶活性的影响 预实验发现12.5 μmol/L AOPDM1为对骨髓间充质干细胞碱性磷酸酶活性影响发生转折的浓度,因此选择附近的4个浓度说明问题。"
2.2.5 AOPDM1对骨髓间充质干细胞成骨分化基因表达的影响 碱性磷酸酶染色及茜素红染色结果表明,12.5 μmol/L是AOPDM1促进成骨的最佳浓度,因此采用12.5 μmol/L浓度进行后续实验。图11显示高糖环境下12.5 μmol/L AOPDM1对骨髓间充质干细胞成骨基因Ⅰ型胶原(成骨诱导7 d)、骨桥蛋白(成骨诱导14 d)、Runx2(成骨诱导7 d)及碱性磷酸酶(成骨诱导7 d)mRNA表达的影响。在不加入AOPDM1的情况下,高糖环境组细胞内Ⅰ型胶原、骨桥蛋白、Runx2及碱性磷酸酶 mRNA的表达均低于正常环境组(P < 0.01,P < 0.001,P < 0.000 1);高糖环境下,加入12.5 μmol/L AOPDM1可提高细胞内Ⅰ型胶原、骨桥蛋白、Runx2及碱性磷酸酶 mRNA的表达(P < 0.05,P < 0.01,P < 0.001,P < 0.000 1)。"
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