PLA/PVP-GEL复合支架对MC3T3-E1细胞生物学行为的影响及成骨性能的初探

口腔生物医学 ›› 2022, Vol. 13 ›› Issue (4): 234-240.

PLA/PVP-GEL复合支架对MC3T3-E1细胞生物学行为的影响及成骨性能的初探

陈安琪¹,吴清霖²,吴勇敏¹,林媛媛¹,赖颖真²

1. 厦门医学院·口腔医学系·口腔生物材料福建省高校工程研究中心
2. 厦门医学院口腔医学系

收稿日期:2022-09-14 修回日期:2022-11-07 出版日期:2022-12-25 发布日期:2022-12-24
通讯作者: 赖颖真 E-mail:dentistyz@126.com
基金资助:
福建省中青年教师教育科研项目（科技类）;大学生创新创业训练计划立项（省级）;2020年厦门医学院自然科学类项目子课题

Effect of PLA/PVP-GEL composite scaffold on the biological behavior and osteogenic properties of MC3T3-E1 cell

Received:2022-09-14 Revised:2022-11-07 Online:2022-12-25 Published:2022-12-24

摘要/Abstract

摘要： 目的：明确聚乳酸（PLA）复合明胶(GEL)与不同比例聚乙烯吡咯烷酮(PVP)复合改性后对成骨细胞MC3T3-E1的生物学行为及成骨性能的影响，为骨组织工程载药方向支架研究奠定研究基础。方法：采用真空冷冻干燥法制备PLA/PVP-GEL复合支架，其中GEL比例固定，PLA/PVP质量比例确定为10:0，8:2，7:3，5:5。利用扫描电镜、红外光谱等进行复合材料理化性能表征，CCK-8法比较各组支架的生物相容性，并通过免疫荧光染色观察MC3T3-E1在各组支架上的黏附形态及分布情况。通过碱性磷酸酶和茜素红染色的定性、定量分析研究各组支架对MC3T3-E1细胞成骨分化的影响。结果：真空冷冻干燥法可制备出具有不规则多孔形貌的复合支架，PLA/GEL复合不同比例的PVP的支架材料亲水性均有改善，MC3T3-E1在各不同质量比例的复合支架上均能黏附，8:2 PLA/PVP-GEL复合支架的细胞黏附形态及黏附率最佳。各组支架均对MC3T3-E1的成骨分化具有积极作用，8:2 PLA/PVP-GEL复合支架对MC3T3-E1的成骨分化促进作用最佳。结论：8:2 PLA/PVP-GEL复合支架可以作为骨组织工程基础载药支架而进行后续载药研究。

关键词: 聚乳酸/聚乙烯吡咯烷酮, 明胶, 复合支架, 成骨分化

Abstract: Objective:To clarify the effects of polylactic acid (PLA) composite gelatin (GEL) modified with different ratios of polyvinylpyrrolidone (PVP) composite on the biological behavior and osteogenic properties of osteoblasts MC3T3-E1, and to lay the research foundation for the study of scaffolds in the direction of drug delivery for bone tissue engineering. Methods:The PLA/PVP-GEL composite scaffold was prepared by vacuum freeze-drying method, in which the mass ratio of PLA/PVP was determined to be 10:0, 8:2, 7:3, 5:5, and the composite volume ratio of GEL to PLA/PVP was fixed at 5%. The physicochemical properties of the composites were characterized using scanning electron microscopy, infrared spectroscopy and contact angle measurement, and the swelling rate, water absorption and porosity of the composite scaffolds were determined. The biocompatibility of each group of scaffolds was compared using CCK-8 method, and the adhesion pattern and distribution of MC3T3-E1 on each group of scaffolds were observed by immunofluorescence staining. The effects of each group of scaffolds on osteogenic differentiation of MC3T3-E1 cells were investigated by qualitative and quantitative analysis of alkaline phosphatase (ALP) and alizarin red staining. Results:The composite scaffolds with irregular porous morphology could be prepared by vacuum freeze-drying method, and the hydrophilicity of scaffolds with different ratios of PLA/GEL composite PVP was improved (P<0.05). MC3T3-E1 could adhere to each composite scaffold with different ratios, and the 8:2 PLA/PVP-GEL composite scaffold had the best cell adhesion morphology and proliferation (P<0.05).The qualitative staining results of ALP and alizarin red were darker in all groups of PLA/PVP-GEL scaffolds compared to the PLA/GEL scaffold group, all groups of scaffolds had positive effects on the osteogenic differentiation of MC3T3-E1, and the 8:2 PLA/PVP-GEL composite scaffold had the best promotion of extracellular mineralization level of MC3T3-E1 (P<0.05). Conclusions:The 8:2 PLA/PVP-GEL composite scaffold can be used as a basic drug-loaded scaffold for bone tissue engineering and subsequent drug loading studies.

Key words: Poly(lactic acid)/poly(vinyl pyrrolidone), Gelatin, Composite scaffold, Osteogenic differentiation

陈安琪吴清霖吴勇敏林媛媛赖颖真. PLA/PVP-GEL复合支架对MC3T3-E1细胞生物学行为的影响及成骨性能的初探[J]. 口腔生物医学, 2022, 13(4): 234-240.

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