载ADM-PLGA纳米微球的ADM-PLGA-NHAC制备及其对人骨肉瘤MG63细胞的抑制作用

杨联军; 荣子杰; 张赞杰; 伍国峰; 张西兵; 王健; 朱立新

doi:10.13418/j.issn.1001-165x.2014.05.018

中国临床解剖学杂志 ›› 2014, Vol. 32 ›› Issue (5) : 585-589. DOI: 10.13418/j.issn.1001-165x.2014.05.018

实验研究

载ADM-PLGA纳米微球的ADM-PLGA-NHAC制备及其对人骨肉瘤MG63细胞的抑制作用

杨联军，　荣子杰，　张赞杰，　伍国峰，　张西兵，　王健，　朱立新

作者信息 +

Synthesis and characterization of nano-hydroxyapatite/collagen scaffolds incorporating ADM-PLGA microspheres and observation of its inhibition on MG63 cells

YANG Lian-jun, RONG Zi-jie, ZHANG Zan-jie, WU Guo-feng, ZHANG Xi-bing, WANG jian, ZHU Li-xin

Author information +

文章历史 +

摘要

目的　研制载阿霉素(ADM)的聚乳酸-羟基乙酸共聚物(PLGA)纳米微球的纳米羟基磷灰石/胶原复合支架（ADM-PLGA-NHAC），研究其性质及体外释药特点，探讨其体外抑制人骨肉瘤MG63细胞的作用，为骨肉瘤的治疗提供新策略。方法　以纳米羟基磷灰石及胶原为原料制备纳米羟基磷灰石/胶原支架并在其中加载ADM-PLGA纳米微球, 通过扫描电子显微镜、体外释放行为等手段评价载药支架材料的性能。以CCK8法、活-死染色评价该复合支架的浸提液在体外对人骨肉瘤MG63细胞株的抗肿瘤活性。结果　复合支架的孔径多在100~200 μm，孔隙率约为82%，微球与支架间结合较为紧密。复合支架具有良好的缓释特性，28 d内能持续缓慢释放阿霉素。复合支架的浸提液对骨肉瘤MG63细胞生长有明显的抑制作用。结论　制备的ADM-PLGA-NHAC复合支架具有良好的药物缓释特性及抗肿瘤效应，是一种具有良好应用前景的抗骨肉瘤骨修复材料。

Abstract

Objective　To study preparation and characteristics of nano-hydroxyapatite/collagen scaffolds incorporating ADM-PLGA microspheres(ADM-PLGA-NHAC) and observe its in-vitro anti-tumor effect. Method　 ADM-PLGA-NHAC were prepared by freezing and lyophylization of composite solution.The characteristic and morphology of the scaffolds were observed by scanning electron microscope, and the release behavior of ADM was investigated in vitro. The antitumor activities of ADM-PLGA-NHAC against human MG63 cell lines were detected by CCK8 colorimetric assay and live/dead cell staining.　Result　The ADM-PLGA microspheres couldbe uniformly distributed in nano-hydroxyapatite/collagen scaffolds to form ADM-PLGA-NHAC scaffolds, which had 100-200 μm pore diameter and over 82% porosity. The release study indicated that the ADM-PLGA-NHAC show sustained release behavior in vitro. The leaching liquor of ADM-PLGA-NHAC could inhibit MG63 cell lines proliferation in a dose-dependent manner.　Conclusion　The ADM-PLGA-NHAC shows sustained release characteristics and anti-tumor effect, and it will have a wide application prospect in the fields of osteosarcoma.

导出引用

杨联军, 荣子杰, 张赞杰, 伍国峰, 张西兵, 王健, 朱立新. 载ADM-PLGA纳米微球的ADM-PLGA-NHAC制备及其对人骨肉瘤MG63细胞的抑制作用[J]. 中国临床解剖学杂志. 2014, 32(5): 585-589 https://doi.org/10.13418/j.issn.1001-165x.2014.05.018

YANG Lian-Jun, RONG Zi-Jie, ZHANG Zan-Jie, WU Guo-Feng, ZHANG Xi-Bing, WANG Jian, SHU Li-Xin. Synthesis and characterization of nano-hydroxyapatite/collagen scaffolds incorporating ADM-PLGA microspheres and observation of its inhibition on MG63 cells[J]. Chinese Journal of Clinical Anatomy. 2014, 32(5): 585-589 https://doi.org/10.13418/j.issn.1001-165x.2014.05.018

中图分类号： R318.08

参考文献

[1] Ferrari S, Smeland S, Mercuri M, et al. Neoadjuvant chemotherapy with high-dose Ifosfamide, high-dose methotrexate, cisplatin, and doxorubicin for patients with localized osteosarcoma of the extremity: a joint study by the Italian and Scandinavian Sarcoma Groups
[J]. J Clin Oncol, 2005,23 (34): 8845-8852.

[2] 杨进城，尹庆水，林骏，等. 自制复合抗肿瘤珊瑚羟基磷灰石人工骨体外抗肿瘤实验
[J]. 中国组织工程研究与临床康复,2007,11(18):3508-3511.

[3] Tahara Y, Ishii Y. Apatite cement containing cis-diammine dichloroplatinum implanted in rabbit femur for sustained release of the anticancer drug and bone formation
[J]. J Orthop Sci, 2001, 6(6): 556-565.

[4] 王健，朱立新，曹延林. 局部药物缓释系统治疗恶性骨肿瘤研究进展
[J]. 广东医学, 2011, (13): 1771-1774.

[5] 王振林，闫玉华，万涛. 羟基磷灰石／胶原类骨仿生复合材料的制备及表征
[J]. 复合材料学报, 2005, 22(2):83-86.

[6] 马宁，曾毅，张莉，等. 多孔纳米羟基磷灰石/胶原复合材料的制备及性能
[J]. 吉林大学学报（医学版）, 2006, 32(1):90-93.

[7] 杨联军，朱立新，包和婧，等. 载阿霉素PLGA纳米微球的制备及其对人骨肉瘤MG63细胞的抑制作用
[J]. 实用医学杂志, 2014, 30(4):528-531.

[8] 刘浩怀，张建华，许庆陵，等. 包埋载药微球的羟基磷灰石/聚氨酯复合组织工程支架的研究
[J]. 无机材料学报, 2011, 26(10):1073-1077.

[9] 林骏，杨进城. 复合抗肿瘤活性珊瑚羟基磷灰石人工骨抗骨巨细胞瘤的体外实验
[J]. 中国组织工程研究与临床康复, 2009, 13(34):6637-6640.

[10]Andronescu E, Ficai A, Albu MG, et al. Collagen-hydroxyapatite/cisplatin drug delivery systems for locoregional treatment of bone cancer
[J]. Technol Cancer Res Treat, 2013, 12(4):275-284.

[11]周雅轩. 纳米载药系统在医药领域中的应用进展
[J]. 天津药学, 2012, 24(1):47-49.

[12]Chavanpatil MD, Patil Y, Panyam J. Susceptibility of nanoparticle-encapsulated paclitaxel to P-glycoprotein-mediated drug efflux
[J]. Int J Pharm, 2006, 320(1-2):150-156.

[13] Barraud L, Merle P, Soma E, et al. Increase of doxorubicin sensitivity by doxorubicin-loading into nanoparticles for hepatocellular carcinoma cells in vitro and in vivo
[J]. J Hepatol, 2005, 42(5):736-743.

[14] Lin Y, Li Y, Ooi C P. 5-Fluorouracil encapsulated HA/PLGA composite microspheres for cancer therapy
[J]. J Mater Sci Mater Med, 2012, 23(10):2453-2460.

[15] Guan WY, Yi WT, Zhi MY, et al. Experimental research on the use of an antineoplastic drug with a bone implant
[J]. Int Orthop, 1990, 14(4):387-391.