期刊导航

论文摘要

聚己内酯—聚乙二醇—血小板浓缩生长因子复合支架的制备及其对人牙周膜干细胞生物学特性的影响

Preparation of polycaprolactone-polyethylene glycol-concentrated growth factor composite scaffolds and the effects on the biological properties of human periodontal ligament stem cells

作者:高丽, 赵明月, 杨顺, 王茹楠, 程佳佳, 陈广生

Author:Gao Li, Zhao Mingyue, Yang Shun, Wang Runan, Cheng Jiajia, Chen Guangsheng

收稿日期:2025-02-06          年卷(期)页码:2025,43(6):819-819-828

期刊名称:华西口腔医学杂志

Journal Name:West China Journal of Stomatology

关键字:血小板浓缩生长因子,聚己内酯—聚乙二醇—血小板浓缩生长因子复合支架,聚己内酯—聚乙二醇支架,人牙周膜干细胞,细胞增殖,成骨分化,

Key words:concentrated growth factor,polycaprolactone-polyethylene glycol-concentrated growth factor composi-te scaffold,polycaprolactone-polyethylene glycol scaffold,human periodontal ligament stem cells,cell proliferation,osteogenic differentiation,

基金项目:贵州省卫健委科学技术基金(gzwkj2023-202);国家自然科学基金(82060204);贵州省科技计划项目[基金黔科合基础—ZK(2021)一般437]

中文摘要

目的 探究负载了血小板浓缩生长因子(CGF)的聚己内酯(PCL)—聚乙二醇(PEG)支架对人牙周膜干细胞(hPDLSCs)黏附、增殖及成骨分化能力的影响。 方法 通过浸入及冻干法制备PCL-PEG-CGF复合支架,观察复合支架的显微结构并检测其机械性能及生物相容性。酶消化法培养hPDLSCs,并通过流式细胞术鉴定hPDLSCs。选取第三代hPDLSCs,分别通过CCK-8实验、4’,6-二脒基-2-苯基吲哚(DAPI)染色、碱性磷酸酶(ALP)染色、茜素红染色及Western blot实验检测PCL-PEG-CGF复合支架对hPDLSCs黏附、增殖及成骨分化能力的影响及成骨相关蛋白[Runt相关转录因子2(Runx2)、ALP、骨形态发生蛋白2(BMP2)]的表达情况。 结果 PCL-PEG-CGF复合支架在扫描电镜下呈孔隙大小不等的蜂窝状结构,其亲水性6 s时θ角接近0°,弹性模量为(4.590 0±0.149 3)MPa,其亲水性、断裂拉伸强度及断裂延伸率均与PCL-PEG支架相似;hPDLSCs在PCL-PEG-CGF复合支架上的黏附能力明显优于PCL-PEG支架(P<0.01),与对照组相比,第3、5、7天时各实验组的hPDLSCs增殖速率显著增加(P<0.01),且PCL-PEG-CGF复合支架与其余组间差异有统计学意义(P<0.01);PCL-PEG-CGF复合支架组ALP活性明显升高(P<0.05),矿化结节数量明显增加及成骨相关蛋白(Runx2、BMP2、ALP)的表达均明显增强(P<0.05)。 结论 PCL-PEG-CGF复合支架具有较好的机械性能及生物相容性,可促进hPDLSCs黏附及增殖,并通过增强成骨相关蛋白的表达促进hPDLSCs的成骨分化能力。

英文摘要

ObjectiveThis study investigated the effects of a polycaprolactone (PCL)-polyethylene glycol (PEG) scaffold incorporated with concentrated growth factor (CGF) on the adhesion, proliferation, and osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs).MethodsThe PCL-PEG-CGF composite scaffold was fabricated using an immersion and freeze-drying technique. Its microstructure, mechanical properties, and biocompatibility were systematically characterized. The hPDLSCs were isolated through enzymatic digestion, and the hPDLSCs were identified through flow cytometry. Third-passage hPDLSCs were seeded onto the composite scaffolds, and their adhesion, proliferation and osteogenic differentiation were assessed using CCK-8 assays, 4’,6-diamidino-2-phenylindole (DAPI) staining, alkaline phosphatase (ALP) staining, alizarin red staining, and Western blot analysis of osteogenesis-related proteins [Runt-related transcription factor 2 (Runx2), ALP, and morphogenetic protein 2 (BMP2)].ResultsScanning electron microscopy revealed that the PCL-PEG-CGF composite scaffold exhibited a honeycomb-like structure with heterogeneous pore sizes. The composite scaffold exhibited excellent hydrophilicity, as evidenced by a contact angle (θ) approaching 0° within 6 s. Its elastic modulus was measured at (4.590 0±0.149 3) MPa, with comparable hydrophilicity, fracture tensile strength, and fracture elongation to PCL-PEG scaffold. The hPDLSCs exhibited significantly improved adhesion to the PCL-PEG-CGF composite scaffold compared with the PCL-PEG scaffold (P<0 .01). additionally, cell proliferation was markedly improved in all the experimental groups on days 3, 5, and 7 (P<0 .01), and statistically significant differences were found between the pcl-peg-cgf group and other groups (P<0 .01). the pcl-peg-cgf group showed significantly elevated alp activity (P<0 .05), increased mineralization nodule formation, and upregulated expression of osteogenic-related proteins (runx2, bmp2 and alp;P<0 .05).ConclusionThe PCL-PEG-CGF composite scaffold exhibited excellent mechanical properties and biocompatibility, enhancing the adhesion and proliferation of hPDLSCs and promoting their osteogenic differentiation by upregulating osteogenic-related proteins.

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