Biocompatibility evaluation of emulsion electrospun nanofibers using osteoblasts for bone tissue engineering
Biocompatibility evaluation of emulsion electrospun nanofibers using osteoblasts for bone tissue engineering
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DOI：
10.1080/09205063.2013.814096
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发表时间：
2013-10
期刊：
影响因子：
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通讯作者：
Lingling Tian;M. Prabhakaran;X. Ding;S. Ramakrishna
中科院分区：
文献类型：
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作者：
Lingling Tian;M. Prabhakaran;X. Ding;S. Ramakrishna
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，在药物包埋方面具有很大的潜力。通过乳液静电纺丝将羟基磷灰石(HA)和层粘连蛋白(Laminin)分别掺入纳米纤维的壳层和芯层，可能有利于支持细胞的黏附、增殖和成熟，而不是单因素包裹的纳米纤维。采用乳液纺丝法制备了聚(L-乳酸-ϵ-己内酯)/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±57 nm。制备的支架的弹性模量在22.7~37.0 Mpa之间。第21天，成骨细胞在PlcL/HA/LAM支架上的细胞增殖率分别高于PlcL/LAM和PlcL/HA支架上的细胞增殖率10.4%和12.0%。第14天碱性磷酸酶检测显示，PLCL/HA/LAM支架上的细胞成熟度显著高于PLCL/HA和PLCL/LAM支架上的碱性磷酸酶活性(p⩽0.05)。在第28天进行的能量色散X射线研究结果也显示，种植在PlcL/HA/LAM支架上的细胞有更高的钙沉积。成骨细胞在PlcL/HA/LAM纳米纤维上黏附、增殖和成熟，促进细胞增殖、碱性磷酸酶活性、骨蛋白表达和矿物质沉积。结果表明，层粘连蛋白和透明质酸分别对成骨细胞的增殖和成熟起作用，并且在新型乳液电纺PlcL/HA/LAM纳米纤维中，两种因素的协同作用增强了成骨细胞的功能，证实了其在骨组织再生中的潜在应用。
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.