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乳液静电纺纳米纤维的生物相容性评价——用于骨组织工程的成骨细胞。

Biocompatibility evaluation of emulsion electrospun nanofibers using osteoblasts for bone tissue engineering.

机构信息

a Key Laboratory of Textile Science & Technology, Ministry of Education of China , Donghua University , Shanghai , 201620 , China .

出版信息

J Biomater Sci Polym Ed. 2013;24(17):1952-68. doi: 10.1080/09205063.2013.814096. Epub 2013 Jul 2.

Abstract

Emulsion electrospinning is an advanced technique to fabricate core-shell structured nanofibrous scaffolds, with great potential for drug encapsulation. Incorporation of dual factors hydroxyapatite (HA) and laminin, respectively, within the shell and core of nanofibers through emulsion electrospinning might be of advantageous in supporting the adhesion, proliferation, and maturation of cells instead of single factor-encapsulated nanofibers. We fabricated poly(L-lactic acid-co-ε-caprolactone) (PLCL)/hydroxyapaptite (PLCL/HA), PLCL/laminin (PLCL/Lam), and PLCL/hydroxyapatite/laminin (PLCL/HA/Lam) scaffolds with fiber diameter of 388 ± 35, 388 ± 81, and 379 ± 57 nm, respectively, by emulsion electrospinning. The elastic modulus of the prepared scaffolds ranged from 22.7-37.0 MPa. The osteoblast proliferation on PLCL/HA/Lam scaffolds, determined on day 21, was found 10.4% and 12.0% higher than the cell proliferation on PLCL/Lam or PLCL/HA scaffold, respectively. Cell maturation determined on day 14, by alkaline phosphatase (ALP) activity, was significantly higher on PLCL/HA/Lam scaffolds than the ALP activity on PLCL/HA and PLCL/Lam scaffolds (p ≤ 0.05). Results of the energy dispersive X-ray studies carried out on day 28 also showed higher calcium deposition by cells seeded on PLCL/HA/Lam scaffolds. Osteoblasts were found to adhere, proliferate, and mature actively on PLCL/HA/Lam nanofibers with enhanced cell proliferation, ALP activity, bone protein expression, and mineral deposition. Based on the results, we can conclude that laminin and HA individually played roles in osteoblast proliferation and maturation, and the synergistic function of both factors within the novel emulsion electrospun PLCL/HA/Lam nanofibers enhanced the functionality of osteoblasts, confirming their potential application in bone tissue regeneration.

摘要

乳液静电纺丝是一种制备核壳结构纳米纤维支架的先进技术,在药物包封方面具有很大的潜力。通过乳液静电纺丝,将双因素羟基磷灰石 (HA) 和层粘连蛋白分别包埋在纳米纤维的壳和核中,可能有利于支持细胞的黏附、增殖和成熟,而不是单一因素包埋的纳米纤维。我们通过乳液静电纺丝制备了聚(L-乳酸-co-ε-己内酯)(PLCL)/羟基磷灰石(PLCL/HA)、PLCL/层粘连蛋白(PLCL/Lam)和 PLCL/羟基磷灰石/层粘连蛋白(PLCL/HA/Lam)支架,纤维直径分别为 388±35、388±81 和 379±57nm,弹性模量为 22.7-37.0MPa。第 21 天,测定的成骨细胞在 PLCL/HA/Lam 支架上的增殖率分别比在 PLCL/Lam 或 PLCL/HA 支架上的细胞增殖率高 10.4%和 12.0%。第 14 天,通过碱性磷酸酶(ALP)活性测定的细胞成熟度在 PLCL/HA/Lam 支架上显著高于 PLCL/HA 和 PLCL/Lam 支架上的 ALP 活性(p≤0.05)。第 28 天进行的能量色散 X 射线研究结果也表明,在 PLCL/HA/Lam 支架上接种的细胞有更高的钙沉积。成骨细胞在 PLCL/HA/Lam 纳米纤维上表现出良好的黏附、增殖和成熟,细胞增殖、ALP 活性、骨蛋白表达和矿物质沉积增强。基于这些结果,我们可以得出结论,层粘连蛋白和 HA 分别在成骨细胞增殖和成熟中发挥作用,而在新型乳液静电纺丝 PLCL/HA/Lam 纳米纤维中两者的协同作用增强了成骨细胞的功能,证实了它们在骨组织再生中的潜在应用。

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