三维打印羟基磷灰石晶须增强复合骨修复支架

doi:10.15541/jim20160628

摘要/Abstract

摘要：

利用三维打印技术成功制备羟基磷灰石晶须(HAPw)增强的聚己内酯(PCL)复合骨修复支架。通过改变三维打印的挤出速度和挤出气压, 使不同含量HAPw均能在PCL基材中一致排列并均匀分布。PCL支架的机械强度随HAPw含量增加显著提高, 添加33wt%HAPw使PCL支架强度提升了高达3倍。此外, HAPw使PCL支架表面与水的接触角从近100º降低至约50º, 有效改善了细胞表面粘附。经过体外人类骨髓间充质干细胞(hBMSC)在支架上的培养实验, 发现添加HAPw的复合支架具有更好的生物相容性, 能够有效促进hBMSC的增殖生长, 且HAPw-PCL复合支架上细胞具有更高的碱性磷酸酶(ALP)活性和OCN、RUNX2等相关成骨基因表达, 显示出hBMSCs向成骨方向更好的分化及成骨活性。

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关键词: 三维打印, 羟基磷灰石晶须, 支架, 骨修复

Abstract:

Development of bioactive scaffolds with controllable architecture and high osteogenic capability is essential for bone tissue engineering. In this study, hydroxyapatite whiskers (HAPw) were added into polycaprolactone (PCL) matrix materials to fabricate scaffolds through 3D printing technique. The HAPw could distribute homogeneously in PCL and in alignment with 3D printing directions by adjusting squeeze parameters. The mechanical strength of PCL-HAPw composite scaffolds increased along with the increase of HAPw content. Adding 33wt% of HAPw remarkably enhanced the compressive strength of PCL scaffolds to 3 times, which can also lower the surface contact angles from 100° of PCL to 50° and thus enhance the surface hydrophilicity. In vitro culturing experiments of human bone marrow mesenchymal cells (hBMSCs) demonstrated that the incorporation of HAPw promoted their bioactive and osteogenic properties, including better cytocompatibility, cell adhesion, proliferation, alkaline phosphatase (ALP) activity, and bone-related gene expressions (OCN, RUNX2). Therefore, 3D-printed HAPw-PCL composite scaffolds showed improved mechanical strength and osteogenesis properties compared to pure PCL scaffolds, and suggest promising applications in bone regeneration.

Key words: 3D printing, hydroxyapatite whiskers, scaffolds, bone regeneration

中图分类号:

TQ174

辛晨, 齐鑫, 朱敏, 赵世昌, 朱钰方. 三维打印羟基磷灰石晶须增强复合骨修复支架[J]. 无机材料学报, 2017, 32(8): 837-844.

XIN Chen, QI Xin, ZHU Min, ZHAO Shi-Chang, ZHU Yu-Fang. Hydroxyapatite Whisker-reinforced Composite Scaffolds Through 3D Printing for Bone Repair[J]. Journal of Inorganic Materials, 2017, 32(8): 837-844.

图/表 11

表1 复合支架编号及打印浆料组成

Table1 Composite scaffold number and composition of printing paste

Samples	HAPw /g	HAPnp /g	PCL /g	CHCl₃ /mL	DMSO /mL
HAPnp-2PCL	0	0.5	1	3.8	0.2
HAPw-2PCL	0.5	0	1	3.8	0.2
HAPw-5PCL	0.2	0	1	3.8	0.2
HAPw-10PCL	0.1	0	1	3.8	0.2
PCL	0	0	1	3.8	0.2

图1 HAPw的不同放大倍数下扫描电镜照片

Fig. 1 SEM images of HAPw at different magnifications

图2 HAPw、PCL 和HAPw-PCL 支架的XRD图谱

Fig. 2 XRD patterns of HAPw, PCL and HAPw-PCL scaffolds

图3 三维打印的PCL(A1, A2), HAPw-10PCL(B1, B2), HAPw-5PCL(C1, C2), HAPw-2PCL(D1, D2)和HAPnp-2PCL支架的SEM照片

Fig. 3 SEM images of 3D-printed PCL(A1,A2), HAPw-10PCL (B1,B2), HAPw-5PCL (C1,C2), HAPw-2PCL (D1,D2) and HAPnp-2PCL scaffolds

图4 PCL、HAPnp-PCL和HAPw-PCL 支架的孔隙率测试结果

Fig. 4 Porosity of PLC, HAPw-PCL and HAPnp-PCL scaffold(*p<0.05 if compared with the PCL scaffold control)

图5 PCL、HAPnp-PCL和HAPw-PCL 支架的抗压能力测试结果

Fig. 5 Compressive strength of the PCL, HAPw-PCL and HAPnp-PCL scaffold(*p<0.05 if compared with the PCL scaffold control)

图6 (A) PCL, HAPw-10PCL, HAPw-5PCL, HAPw-2PCL和 HAPnp-2PCL复合支架表面水滴在不同时间(0、10、20 min)的状态, (B)复合支架表面接触角室内空气下随时间的变化

Fig. 6 (A) Water contact angle of PCL scaffolds (first panel), HAPw-10PCL scaffolds (second panel), HAPw-5PCL scaffolds (third panel), HAPw-2PCL scaffolds (fourth panel), and HAPnp-2PCL. Ascending contact angles measured at varying times (0, 10, and 20 min), and (B) quantification of the contact angle with time

图7 hBMSCs在PCL 和HAPw-PCL 复合支架上培养1、3、7 d后的细胞增殖情况

Fig. 7 Proliferation of hBMSCs on the PCL and HAPw-PCL scaffolds for 1, 3 and 7 d (*p<0.05 if compared with the PCL scaffold control)

图8 hBMSCs在PCL 和HAPw-PCL 复合支架上培养7、14 d后的碱性磷酸酶活性结果

Fig. 8 ALP activity of hBMSCs cultured on PCL and HAPw-PCL scaffolds for 7 and 14 d (*p<0.05 if compared with the PCL scaffold control)

图9 hBMSCs在HAPw-PCL复合支架上培养7 d和14 d后的相关成骨基因表达(ALP(A), RUNX2(B), OCN(C))

Fig. 9 Osteogenic expression of ALP (A), OCN (B), RUNX2 (C) for hBMSCs cultured on the PCL and HAPw-PCL scaffolds by qRT-PCR analysis after 7 d and 14 d (*p < 0.05 if compared with the PCL scaffold control)

图10 hBMSCs 在PCL(A)、HAPw-10PCL (B)、HAPw-5PCL (C) 和 HAPw-2PCL (D)支架上培养21d后的ECM矿化情况

Fig. 10 ECM mineralization of hBMSCs on the PCL(A), HAPw-10PCL (B), HAPw-5PCL (C) and HAPw-2PCL (D) scaffolds after culturing for 21 d

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