包封骨形态发生蛋白2的不同相对分子质量聚乳酸-聚乙醇酸共聚物微囊促进成骨效果的研究

doi:10.7518/hxkq.2024075

华西口腔医学杂志 ›› 2024, Vol. 42 ›› Issue (5): 572-580.doi: 10.7518/hxkq.2024075

包封骨形态发生蛋白2的不同相对分子质量聚乳酸-聚乙醇酸共聚物微囊促进成骨效果的研究

袁莉红¹^,²^,³(), 陈晨¹^,²^,³, 马语笛²^,³, 梁睿贞²^,³^,⁴()

^1.南京医科大学附属口腔医院牙体牙髓科，南京 210029
^2.口腔疾病研究与防治国家级重点实验室培育建设点，南京 210029
^3.江苏省口腔转化医学工程研究中心，南京 210029
^4.南京医科大学附属口腔医院第七门诊部，南京 210029

收稿日期:2024-03-01 修回日期:2024-05-17 出版日期:2024-10-01 发布日期:2024-09-21
通讯作者: 梁睿贞 E-mail:1449672307@qq.com;13958432@qq.com
作者简介:袁莉红，医师，硕士，E-mail：1449672307@qq.com
基金资助:
国家自然科学基金(81970927);江苏省重点研发计划社会发展项目(BE2022797)

Osteogenic effect of poly(lactic-co-glycolic acid) microcapsules with different molecular weights encapsulating bone morphogenetic protein 2

Yuan Lihong¹^,²^,³(), Chen Chen¹^,²^,³, Ma Yudi²^,³, Liang Ruizhen²^,³^,⁴()

^1.Dept. of Endodontics, the Affiliated Stomatological Hospital of Nanjing Medical University, Nanjing 210029, China
^2.State Key Laboratory Cultivation Base of Research, Prevention and Treatment for Oral Diseases, Nanjing 210029, China
^3.Jiangsu Province Engineering Research Center of Stomatological Translational Medicine, Nanjing 210029, China
^4.The Seventh Clinic, the Affiliated Stomatological Hospital of Nanjing Medical University, Nanjing 210029, China

Received:2024-03-01 Revised:2024-05-17 Online:2024-10-01 Published:2024-09-21
Contact: Liang Ruizhen E-mail:1449672307@qq.com;13958432@qq.com
Supported by:
National Natural Science Foundation of China(81970927);Key Research and Development Program (Social Development Project) of Jiangsu Province(BE2022797)

摘要/Abstract

摘要：

目的探索不同相对分子质量聚乳酸-聚乙醇酸共聚物（PLGA）微囊包封骨形态发生蛋白2（BMP-2）对成骨细胞骨形成能力的影响。方法用双通道微量注射泵制备包封BMP-2的2种相对分子质量（12 000和30 000）的PLGA微囊。用光学显微镜及扫描电子显微镜（SEM）观察微囊形态结构；磷酸盐缓冲溶液浸泡法表征微囊缓释性能；细胞Calcein-AM/PI染色及CCK-8法检测微囊细胞相容性；Transwell迁移实验检测包封BMP-2的微囊作用48 h对MC3T3-E1细胞的趋化作用；碱性磷酸酶活力测定、茜素红染色法检测微囊作用MC3T3-E1细胞后对细胞骨形成能力的影响。结果 2种相对分子质量的微囊均表面光滑，具有良好的细胞相容性。相对分子质量12 000的微囊的趋化作用最佳。相对分子质量30 000的微囊较相对分子质量12 000的微囊缓释时间长，且初始爆发量减少了约25%。成骨诱导14、21 d后，相对分子质量30 000的微囊形成的钙沉积结节较相对分子质量12 000的微囊多。结论本研究通过调控PLGA的相对分子质量控制BMP-2的释放，发现相对分子质量30 000的微囊能够更好地诱导MC3T3-E1细胞的长期骨形成能力。

关键词: 聚乳酸-聚乙醇酸共聚物微囊, 骨形态发生蛋白2, 成骨

Abstract:

Objective This study aimed to explore the effects of bone morphogenetic protein 2 (BMP-2) encapsula-ted in poly(lactic-co-glycolic acid) (PLGA) microcapsules with different molecular weights on the osteogenic ability of osteoblasts. Methods PLGA microcapsules with different molecular weights (12 000, 30 000) encapsulating BMP-2, were prepared using a dual-channel microinjection pump. The morphology and structure of the microcapsules were characterized by optical microscopy and scanning electron microscopy. The sustained-release performance of the microcapsules was characterized by phosphate buffered saline immersion method. The cell compatibility of the microcapsules was detected by the Calcein-AM/PI staining and CCK-8 method. The chemotactic effect of BMP-2-encapsulated microcapsules on MC3T3-E1 cells after 48 h of treatment was detected by the Transwell assay. The alkaline phosphatase activity assay and Alizarin Red S staining were used to characterize the effect of microcapsules on the osteogenic ability of MC3T3-E1 cells. Results Both types of microcapsules with different molecular weights exhibited smooth surfaces, as well as uniform and good cell compatibility. The chemotactic effect of the 12 000 microcapsules was outstanding. The 30 000 microcapsules had a longer sustained-release time, and the initial burst release was reduced by approximately 25% compared with the 12 000 microcapsules. In addition, 30 000 microcapsules performed better in long-term osteogenesis induction than 12 000 microcapsules. Conclusion In this study, the release of BMP-2 is regulated by adjusting the molecular weight of PLGA, and the results indicate that 30 000 microcapsules can better induce the long-term osteogenic ability of MC3T3-E1 cells.

Key words: poly(lactic-co-glycolic acid) microcapsules, bone morphogenetic protein 2, osteogenesis

中图分类号:

R318.08

袁莉红, 陈晨, 马语笛, 梁睿贞. 包封骨形态发生蛋白2的不同相对分子质量聚乳酸-聚乙醇酸共聚物微囊促进成骨效果的研究[J]. 华西口腔医学杂志, 2024, 42(5): 572-580.

Yuan Lihong, Chen Chen, Ma Yudi, Liang Ruizhen. Osteogenic effect of poly(lactic-co-glycolic acid) microcapsules with different molecular weights encapsulating bone morphogenetic protein 2[J]. West China Journal of Stomatology, 2024, 42(5): 572-580.

图/表 6

图 1

图 2

图 3

图 4

图 5

图 6

参考文献 19

1	Ball JR, Shelby T, Hernandez F, et al. Delivery of grow-th factors to enhance bone repair[J]. Bioengineering (Basel), 2023, 10(11): 1252.
2	Wei J, Xia X, Xiao S, et al. Sequential dual-biofactor release from the scaffold of mesoporous HA microspheres and PLGA matrix for boosting endogenous bone regeneration[J]. Adv Healthc Mater, 2023, 12(20): e2300624.
3	Minardi S, Fernandez-Moure JS, Fan D, et al. Biocompatible PLGA-mesoporous silicon microspheres for the controlled release of BMP-2 for bone augmentation[J]. Pharmaceutics, 2020, 12(2): 118.
4	De Witte TM, Fratila-Apachitei LE, Zadpoor AA, et al. Bone tissue engineering via growth factor delivery: from scaffolds to complex matrices[J]. Regen Biomater, 2018, 5(4): 197-211.
5	Guo X, Song P, Li F, et al. Research progress of design drugs and composite biomaterials in bone tissue engineering[J]. Int J Nanomedicine, 2023, 18: 3595-3622.
6	Shah NJ, Macdonald ML, Beben YM, et al. Tunable dual growth factor delivery from polyelectrolyte multilayer films[J]. Biomaterials, 2011, 32(26): 6183-6193.
7	Minardi S, Pandolfi L, Taraballi F, et al. PLGA-mesoporous silicon microspheres for the in vivo controlled temporospatial delivery of proteins[J]. ACS Appl Mater Interfaces, 2015, 7(30): 16364-16373.
8	Jin S, Xia X, Huang J, et al. Recent advances in PLGA-based biomaterials for bone tissue regeneration[J]. Acta Biomater, 2021, 127: 56-79.
9	Sun F, Sun X, Wang H, et al. Application of 3D-printed, PLGA-based scaffolds in bone tissue engineering[J]. Int J Mol Sci, 2022, 23(10): 5831.
10	Wang Y, Zhao L, Zhou L, et al. Sequential release of vascular endothelial growth factor-A and bone morphogenetic protein-2 from osteogenic scaffolds assembled by PLGA microcapsules: a preliminary study in vitro [J]. Int J Biol Macromol, 2023, 232: 123330.
11	王莹, 陈晨, 陈刚. BMP-2缓释型PLGA微囊作为引导骨再生支架的初步研究[J]. 南京医科大学学报(自然科学版), 2022, 42(9): 1216-1222.
	Wang Y, Chen C, Chen G. Preliminary study of a BMP-2 releasing scaffold comprised of PLGA microspheres for guiding bone regeneration[J]. J Nanjing Med Univ (Nat Sci), 2022, 42(9): 1216-1222.
12	Nyberg E, Holmes C, Witham T, et al. Growth factor-eluting technologies for bone tissue engineering[J]. Drug Deliv Transl Res, 2016, 6(2): 184-194.
13	Wani TU, Khan RS, Rather AH, et al. Local dual delive-ry therapeutic strategies: using biomaterials for advan-ced bone tissue regeneration[J]. J Control Release, 2021, 339: 143-155.
14	Halloran D, Durbano HW, Nohe A. Bone morphogenetic protein-2 in development and bone homeostasis[J]. J Dev Biol, 2020, 8(3): 19.
15	Shi J, Dai W, Gupta A, et al. Frontiers of hydroxyapatite composites in bionic bone tissue engineering[J]. Materials (Basel), 2022, 15(23): 8475.
16	Rmaidi A, Zelzer M, Sindji L, et al. Impact of the physico-chemical properties of polymeric microspheres functionalized with cell adhesion molecules on the behavior of mesenchymal stromal cells[J]. Mater Sci Eng C Mater Biol Appl, 2021, 121: 111852.
17	Zhao D, Zhu T, Li J, et al. Poly(lactic-co-glycolic acid)-based composite bone-substitute materials[J]. Bioact Ma-ter, 2020, 6(2): 346-360.
18	Lee SK, Han CM, Park W, et al. Synergistically enhan-ced osteoconductivity and anti-inflammation of PLGA/β-TCP/Mg(OH)₂ composite for orthopedic applications[J]. Mater Sci Eng C Mater Biol Appl, 2019, 94: 65-75.
19	Yuan Y, Xu Y, Mao Y, et al. Three birds, one stone: an os-teo-microenvironment stage-regulative scaffold for bone defect repair through modulating early osteo-immunomodulation, middle neovascularization, and later osteogenesis[J]. Adv Sci (Weinh), 2024, 11(6): e2306428.

[1]	董丹妮, 黄艳玲, 赖颖真, 尹戈. 钛表面不同改性处理后修饰胶原对体外成骨性能影响的比较研究[J]. 华西口腔医学杂志, 2024, 42(4): 452-461.
[2]	胡静, 崔智华, 黄克强, 苏荣健, 赵颂. GRP78-c-Src信号通路介导周期性牵张力作用下牙周膜成纤维细胞成骨分化的机制研究[J]. 华西口腔医学杂志, 2024, 42(3): 304-312.
[3]	孙艳萍, 廖立. 表面纳米形貌对牙周膜干细胞衰老的作用研究[J]. 华西口腔医学杂志, 2024, 42(2): 172-180.
[4]	唐小雪, 周政, 李启期, 姜丹丹. 西格列汀激活基质细胞衍生因子-1/CXC趋化因子受体4信号通路对脂多糖诱导的人牙周膜干细胞增殖、凋亡、炎症和成骨分化的影响[J]. 华西口腔医学杂志, 2024, 42(1): 37-45.
[5]	李胜鸿, 彭世元, 罗小玲, 王奕佩, 徐晓梅. 柚皮素通过基质细胞衍生因子1/趋化因子受体4信号轴对脂多糖作用下人牙周膜干细胞抗炎、成血管和成骨分化能力的影响[J]. 华西口腔医学杂志, 2023, 41(2): 175-184.
[6]	张鹏, 左东川, 牟思宇, 钟宇彤, 袁小平, 曾锦. 内向整流钾2.1通道蛋白对人牙囊细胞成骨分化的影响及其机制研究[J]. 华西口腔医学杂志, 2022, 40(2): 139-147.
[7]	石玉. 能量代谢在成骨和破骨细胞中的研究[J]. 华西口腔医学杂志, 2021, 39(5): 501-509.
[8]	庞鸣, 韦红霞, 陈茜. 长链非编码RNA钾离子电压门控通道亚家族Q成员1重叠转录本1通过靶向miR-24-3p调控人牙周膜干细胞增殖和成骨分化[J]. 华西口腔医学杂志, 2021, 39(5): 547-554.
[9]	牟婷琛, 冯剑颖. 软骨干细胞标记及诱导分化的研究进展[J]. 华西口腔医学杂志, 2021, 39(1): 108-114.
[10]	谢旭东, 赵蕾, 吴亚菲, 王骏. 骨形态发生蛋白1/tolloid蛋白酶家族在牙及骨组织发育中的作用[J]. 华西口腔医学杂志, 2020, 38(5): 589-593.
[11]	吴湘楠, 马媛媛, 浩志超, 王航. 溶血磷脂酸对骨组织细胞生物学调控功能的研究进展[J]. 华西口腔医学杂志, 2020, 38(3): 324-329.
[12]	温佳慧, 吴燕岷, 陈莉丽. 非编码RNA在人牙周膜来源细胞成骨分化中的作用[J]. 华西口腔医学杂志, 2020, 38(3): 330-337.
[13]	林永盛,王丰芝,雷晓静,何健民. 基质细胞衍生因子-1对炎症和正常来源的人牙周膜干细胞成骨分化能力的影响[J]. 华西口腔医学杂志, 2019, 37(5): 469-475.
[14]	孟姝,林意蓝,赵蕾,徐屹. 正畸治疗与牙周硬组织健康的关系及处理[J]. 华西口腔医学杂志, 2019, 37(4): 343-349.
[15]	李颉颃,苏志飞,白璇,袁鹤,李继遥. 唑来膦酸对大鼠骨髓间充质干细胞增殖及成骨分化的作用研究[J]. 华西口腔医学杂志, 2019, 37(3): 242-247.

包封骨形态发生蛋白2的不同相对分子质量聚乳酸-聚乙醇酸共聚物微囊促进成骨效果的研究

Osteogenic effect of poly(lactic-co-glycolic acid) microcapsules with different molecular weights encapsulating bone morphogenetic protein 2

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

图/表 6

参考文献 19

相关文章 15

编辑推荐

Metrics

本文评价