Lai Guo-Jyun, Shalumon K T, Chen Jyh-Ping
Department of Chemical and Materials Engineering, Chang Gung University of Science and Technology, Taoyuan, Taiwan, Republic of China.
Department of Chemical and Materials Engineering, Chang Gung University of Science and Technology, Taoyuan, Taiwan, Republic of China ; Research Center for Industry of Human Ecology, Chang Gung University of Science and Technology, Taoyuan, Taiwan, Republic of China.
Int J Nanomedicine. 2015 Jan 12;10:567-84. doi: 10.2147/IJN.S73780. eCollection 2015.
Incorporation of nanohydroxyapatite (nHAP) within a chitosan (CS)/silk fibroin (SF) nanofibrous membrane scaffold (NMS) may provide a favorable microenvironment that more closely mimics the natural bone tissue physiology and facilitates enhanced osteogensis of the implanted cell population. In this study, we prepared pristine CS/SF NMS, composite CS/SF/nHAP NMS containing intrafibrillar nHAP by in situ blending of 10% or 30% nHAP before the electrospinning step, and composite CS/SF/nHAP NMS containing extrafibrillar nHAP by depositing 30% nHAP through alternative soaking surface mineralization. We investigated the effect of the incorporation of HAP nanoparticles on the physicochemical properties of pristine and composite NMS. We confirmed the presence of ~30 nm nHAP in the composite nanofibrous membranes by thermogravimetry analysis (TGA), X-ray diffraction (XRD), and scanning electron microscopy (SEM), either embedded in or exposed on the nanofiber. Nonetheless, the alternative soaking surface mineralization method drastically influenced the mechanical properties of the NMS with 88% and 94% drop in Young's modulus and ultimate maximum stress. Using in vitro cell culture experiments, we investigated the effects of nHAP content and location on proliferation and osteogenic differentiation of human bone marrow mesenchymal stem cells (hMSCs). The proliferation of hMSCs showed no significant difference among pristine and composite NMS. However, the extent of osteogenic differentiation of hMSCs was found to be positively correlated with the content of nHAP in the NMS, while its location within the nanofiber played a less significant role. In vivo experiments were carried out with hMSCs seeded in CS/SF/30%nHAP NMS prepared by in situ blending and subcutaneous implantation in nude mice. Micro-computed tomography images as well as histological and immunohistochemical analysis of the retrieved hMSCs/NMS construct 1 and 2 months postimplantation indicated that NMS had the potential for bone regeneration and can be suggested as a promising scaffold for bone tissue engineering.
将纳米羟基磷灰石（nHAP）掺入壳聚糖（CS）/丝素蛋白（SF）纳米纤维膜支架（NMS）中，可能会提供一个更接近天然骨组织生理状态的良好微环境，并促进植入细胞群体的骨生成增强。在本研究中，我们制备了原始的CS/SF NMS、通过在静电纺丝步骤前原位共混10%或30%的nHAP而含有原纤维内nHAP的复合CS/SF/nHAP NMS，以及通过交替浸泡表面矿化沉积30%的nHAP而含有原纤维外nHAP的复合CS/SF/nHAP NMS。我们研究了掺入HAP纳米颗粒对原始和复合NMS物理化学性质的影响。通过热重分析（TGA）、X射线衍射（XRD）和扫描电子显微镜（SEM），我们证实了在复合纳米纤维膜中存在约30 nm的nHAP，其要么嵌入纳米纤维中，要么暴露在纳米纤维上。尽管如此，交替浸泡表面矿化方法极大地影响了NMS的力学性能，杨氏模量和极限最大应力分别下降了88%和94%。通过体外细胞培养实验，我们研究了nHAP含量和位置对人骨髓间充质干细胞（hMSCs）增殖和成骨分化的影响。hMSCs的增殖在原始和复合NMS之间没有显著差异。然而，发现hMSCs的成骨分化程度与NMS中nHAP含量呈正相关，而其在纳米纤维内的位置作用较小。用通过原位共混制备并皮下植入裸鼠的CS/SF/30%nHAP NMS接种hMSCs进行体内实验。植入后1个月和2个月对取出的hMSCs/NMS构建体进行的微型计算机断层扫描图像以及组织学和免疫组织化学分析表明，NMS具有骨再生潜力，可被认为是一种有前途的骨组织工程支架。
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