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制备载 BMP-9 和 P-15 肽水凝胶的 3D 打印 PLGA 支架及其在兔骨缺损治疗中的应用。

Preparation of 3D Printing PLGA Scaffold with BMP-9 and P-15 Peptide Hydrogel and Its Application in the Treatment of Bone Defects in Rabbits.

机构信息

Department of Orthopedics, The Second Affiliated Hospital of Nanchang University, Nanchang 330006, China.

Department of Orthopedics, Xiangzhou District People's Hospital, Xiangyang 441000, China.

出版信息

Contrast Media Mol Imaging. 2022 Jul 31;2022:1081957. doi: 10.1155/2022/1081957. eCollection 2022.

Abstract

OBJECTIVE

To prepare a three-dimensional (3D) printing polylactic acid glycolic acid (PLGA) scaffold with bone morphogenetic protein-9 (BMP-9) and P-15 peptide hydrogel and evaluate its application in treating bone defects in rabbits.

METHODS

3D printing PLGA scaffolds were formed and scanned by electron microscopy. Their X-ray diffraction (XRD), degradation, and compressive strength were characterized. BMP-9 and P-15 hydrogels were prepared. Flow cytometry was used to detect apoptosis, and an electron microscope was used to evaluate cell adhesion to scaffolds. Alkaline phosphatase (ALP), type 1 collagen (Col-I), osteocalcin (OCN), runt-related transcription factor 2 (RUNX2), and osterix (SP7) were detected by western blotting. MicroCT was used to detect new bone formation, and bone tissue-related protein expressions were determined in the rabbit model with bone defects.

RESULTS

The 3D printing scaffolds were cylindrical, and the inner diameter of the scaffolds was about 1 mm. The bread peak with wide distribution showed that the 3D printing only involved a physical change, which did not change the properties of the materials. The degradation rate of scaffolds was 9.38%, which met the requirements of properties of biological scaffolds. The water absorption of the support was about 9.09%, and the compressive strength was 15.83 N/mm. In the coculture of bone marrow mesenchymal stem cells (BMSCs) with scaffolds, the 2% polypeptide hydrogel showed the most obvious activity in promoting the differentiation of BMSCs. Flow cytometry showed that the 0% and 2% groups did not cause obvious apoptosis compared with the control group. Scaffolds with 2% and 4% polypeptide promoted the expression of ALP, COL-1, OCN, RUNX2, and Sp7 in BMSCs. experiments showed that the expression of ALP, COL-1, OCN, RUNX2, and Sp7 protein in the 2% polypeptide scaffold group increased significantly compared with the model group. MicroCT detection demonstrated that the 2% polypeptide scaffold had good bone repair ability.

CONCLUSION

The PLGA scaffolds combined with BMP-9 and P-15 peptide hydrogels had good biological and mechanical properties and could repair bone defects in rabbits.

摘要

目的

制备载有骨形态发生蛋白-9(BMP-9)和 P-15 肽水凝胶的三维(3D)打印聚乳酸-羟基乙酸共聚物(PLGA)支架,并评估其在治疗兔骨缺损中的应用。

方法

通过电子显微镜形成和扫描 3D 打印的 PLGA 支架。对其 X 射线衍射(XRD)、降解和压缩强度进行了表征。制备 BMP-9 和 P-15 水凝胶。通过流式细胞术检测细胞凋亡,电子显微镜评估细胞对支架的黏附。通过蛋白质印迹法检测碱性磷酸酶(ALP)、I 型胶原(Col-I)、骨钙素(OCN)、 runt 相关转录因子 2(RUNX2)和成骨转录因子(SP7)。采用微 CT 检测新骨形成,并在兔骨缺损模型中检测骨组织相关蛋白的表达。

结果

3D 打印支架呈圆柱形,支架内径约 1mm。宽分布的面包峰表明 3D 打印只涉及物理变化,并未改变材料的性质。支架的降解率为 9.38%,符合生物支架性能要求。支架的吸水率约为 9.09%,压缩强度为 15.83N/mm。在骨髓间充质干细胞(BMSCs)与支架的共培养中,2%多肽水凝胶在促进 BMSCs 分化方面表现出最明显的活性。流式细胞术显示,0%和 2%组与对照组相比,未引起明显的细胞凋亡。2%和 4%多肽支架促进了 BMSCs 中 ALP、COL-1、OCN、RUNX2 和 Sp7 的表达。实验表明,2%多肽支架组中 ALP、COL-1、OCN、RUNX2 和 Sp7 蛋白的表达明显高于模型组。微 CT 检测表明,2%多肽支架具有良好的骨修复能力。

结论

PLGA 支架结合 BMP-9 和 P-15 肽水凝胶具有良好的生物力学性能,可修复兔骨缺损。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a1d4/9357721/0dd49731a2bd/CMMI2022-1081957.001.jpg

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