Yu Xiang, Shen Gengyang, Shang Qi, Zhang Zhida, Zhao Wenhua, Zhang Peng, Liang De, Ren Hui, Jiang Xiaobing
Department of Spinal Surgery, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, China.
The First Clinical College of Guangzhou University of Chinese Medicine, Guangzhou, China.
Int J Biol Macromol. 2021 Dec 15;193(Pt A):510-518. doi: 10.1016/j.ijbiomac.2021.10.036. Epub 2021 Oct 25.
In this study, we investigated the effect of three-dimensional of naringin/gelatin microspheres/nano-hydroxyapatite/silk fibroin (NG/GMs/nHA/SF) scaffolds on repair of a critical-size bone defect of lumbar 6 in osteoporotic rats. In this work, a cell-free scaffold for bone-tissue engineering based on a silk fibroin (SF)/nano-hydroxyapatite (nHA) scaffold was developed. The scaffold was fabricated by lyophilization. Naringin (NG) was loaded into gelatin microspheres (GMs), which were encapsulated in the nHA/SF scaffolds. The materials were characterized using x ray diffraction, Fourier-transform infrared spectroscopy, scanning electron microscopy, transmission electron microscopy and thermogravimetric analysis. Moreover, the biomechanics, degradation, and drug-release profile of the scaffold were also evaluated. In vitro, the effect of the scaffold on the adhesion, proliferation, and osteogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs) was evaluated. In vivo, at 3 months after ovariectomy, a critical-size lumbar defect was indued in the rats to evaluate scaffold therapeutic potential. A 3-mm defect in L6 developed in 60 SD rats, which were randomly divided into SF scaffold, nHA/SF scaffold, NG/nHA/SF scaffold, NG/GMs/nHA/SF scaffold, and blank groups (n = 12 each). At 4, 8, 12, and 16 weeks postoperatively, osteogenesis was evaluated by X-ray, micro-computed tomography, hematoxylin-eosin staining, and fast green staining, and by analysis of BMP-2, Runx2, and Ocn protein levels at 16 weeks. In our results, NG/GM/nHA/SF scaffolds exhibited good biocompatibility, biomechanical strength, and promoted BMSC adhesion, proliferation, and calcium nodule formation in vitro. Moreover, NG/GMs/nHA/SF scaffolds showed greater osteogenic differentiation potential than the other scaffolds in vitro. In vivo, gradual new bone formation was observed, and bone defects recovered by 16 weeks in the experimental group. In the blank group, limited bone formation was observed, and the bone defect was obvious. In conclusion, NG/GMs/nHA/SF scaffolds promoted repair of a lumbar 6 defect in osteoporotic rats. Therefore, the NG/GMs/nHA/SF biocomposite scaffold has potential as a bone-defect-filling biomaterial for bone regeneration.
在本研究中，我们调查了柚皮苷/明胶微球/纳米羟基磷灰石/丝素蛋白（NG/GMs/nHA/SF）三维支架对骨质疏松大鼠腰6临界尺寸骨缺损修复的影响。在这项工作中，开发了一种基于丝素蛋白（SF）/纳米羟基磷灰石（nHA）支架的用于骨组织工程的无细胞支架。该支架通过冻干法制备。将柚皮苷（NG）负载到明胶微球（GMs）中，然后将其包裹在nHA/SF支架中。使用X射线衍射、傅里叶变换红外光谱、扫描电子显微镜、透射电子显微镜和热重分析对材料进行表征。此外，还评估了支架的生物力学性能、降解情况和药物释放曲线。在体外，评估了该支架对大鼠骨髓间充质干细胞（BMSCs）黏附、增殖和成骨分化的影响。在体内，卵巢切除术后3个月，在大鼠中诱导出临界尺寸的腰椎缺损，以评估支架的治疗潜力。60只SD大鼠的L6处形成3毫米的缺损，将其随机分为SF支架组、nHA/SF支架组、NG/nHA/SF支架组、NG/GMs/nHA/SF支架组和空白组（每组n = 12）。术后4、8、12和16周，通过X射线、显微计算机断层扫描、苏木精-伊红染色和固绿染色，以及在16周时分析BMP-2、Runx2和Ocn蛋白水平来评估成骨情况。在我们的结果中，NG/GM/nHA/SF支架表现出良好的生物相容性、生物力学强度，并在体外促进了BMSC的黏附、增殖和钙结节形成。此外，NG/GMs/nHA/SF支架在体外显示出比其他支架更大的成骨分化潜力。在体内，观察到实验组逐渐有新骨形成，到16周时骨缺损得到修复。在空白组中，观察到有限的骨形成，骨缺损明显。总之，NG/GMs/nHA/SF支架促进了骨质疏松大鼠腰6缺损的修复。因此，NG/GMs/nHA/SF生物复合支架有潜力作为一种用于骨再生的骨缺损填充生物材料。
