Osteogenesis of bone marrow mesenchymal stem cells on hydroxyapatite/icariin/poly(lactic-co-glycolic acid) scaffolds

doi:10.3969/j.issn.2095-4344.2083

Abstract

Abstract:

BACKGROUND: Bone tissue engineering has provided a novel ideal for treating bone defects in clinic. This study is the first to combine traditional Chinese medicine with the nanostructures of tissue-engineered scaffolds in order to explore and construct a new bone tissue substitute material for the treatment of bone defects.

OBJECTIVE: To investigate the osteogenic activity of icariin (ICA)/hydroxyapatite (HA)/poly(lactic-co-glycolic acid) (PLGA) composite scaffolds.

METHODS: A HA/PLGA composite scaffold was prepared by physical blending of HA and PLGA, and was then soaked in ICA solution of different concentrations to obtain the HA/ICA/PLGA scaffold. Rabbit bone marrow mesenchymal stem cells were used to evaluate the cell adhesion, proliferation, osteogenesis and cytotoxicity of the composite scaffold. The cell adhesion, proliferation and cytotoxicity were detected by MTT method. The activities of alkaline phosphatase and osteocalcin were detected by ELISA. The expression levels of osteogenic genes and proteins were detected by fluorescence quantitative PCR and western blot assay, respectively.

RESULTS AND CONCLUSION: Adding appropriate amount of HA into PLGA could improve the mechanical strength of the scaffold, and 10% HA had the best effect with tensile strength of (1.67±0.37) MPa, and compression modulus of (4.17±1.62) MPa, and nanostructure would be formed on the surface of the scaffold. The nanostructure could promote the adhesion of bone marrow mesenchymal stem cells on the surface of the scaffold. ICA did not affect the proliferation of bone marrow mesenchymal stem cells on the composite scaffold. However, the HA/PLGA composite scaffold soaked in 1.00 µmol/L ICA aqueous solution had the optimal osteogenic differentiation function, and the expression levels of alkaline phosphatase, osteocalcin, osteogenic related genes and proteins (Runx-2 and COL I) were increased. The ICA/HA/PLGA scaffold had no cytotoxicity. These results suggest that HA (10%)/ICA (1.00 µmol/L)/PLGA scaffold has good mechanical properties, osteogenesis and biocompatibility, which has the potential to be a favorable scaffold for bone tissue engineering.

Key words:

tissue engineering scaffolds, poly(lactic-co-glycolic acid), hydroxyapatite, icariin, bone defects, osteogenic induction

CLC Number:

Wang Dexin, Xu Zhanwu, Pei Guoxian.

Osteogenesis of bone marrow mesenchymal stem cells on hydroxyapatite/icariin/poly(lactic-co-glycolic acid) scaffolds [J]. Chinese Journal of Tissue Engineering Research, 2020, 24(25): 3974-3980.

Figures/Tables 9

References

[1] FERNANDEZ DE GRADO G, KELLER L, IDOUX-GILLET Y, et al. Bone substitutes: a review of their characteristics, clinical use, and perspectives for large bone defects management. J Tissue Eng. 2018;9:2041731418776819.

[2] MORELLI I, DRAGO L, GEORGE DA, et al. Managing large bone defects in children: a systematic review of the 'induced membrane technique'. J Pediatr Orthop B. 2018;27(5): 443-455.

[3] LESENSKY J, PRINCE DE. Distraction osteogenesis reconstruction of large segmental bone defects after primary tumor resection: pitfalls and benefits. Eur J Orthop Surg Traumatol. 2017;27(6):715-727.

[4] WANG S, ZHAO Z, YANG Y, et al. A high-strength mineralized collagen bone scaffold for large-sized cranial bone defect repair in sheep. Regen Biomater. 2018;5(5):283-292.

[5] CHU LY, JIANG GQ, HU XL, et al. Osteogenesis, vascularization and osseointegration of a bioactive multiphase macroporous scaffold in the treatment of large bone defects. J Mater Chem B. 2018; 6(25): 4197-4204.

[6] PEÑAFLOR T, CHAI Y, TAGAYA M. Hydroxyapatite Nanoparticle Coating on Polymer for Constructing Effective Biointeractive Interfaces. Journal of Nanomaterials.2019;(3): 1-23.

[7] SIDDIQUI HA, PICKERING KL, MUCALO MR. A Review on the Use of Hydroxyapatite-Carbonaceous Structure Composites in Bone Replacement Materials for Strengthening Purposes. Materials (Basel). 2018;11(10): E1813.

[8] JUN SH, LEE EJ, JANG TS, et al. Bone morphogenic protein-2 (BMP-2) loaded hybrid coating on porous hydroxyapatite scaffolds for bone tissue engineering. J Mater Sci Mater Med. 2013;24(3):773-782.

[9] CHEN B, NIU SP, WANG ZY, et al. Local administration of icariin contributes to peripheral nerve regeneration and functional recovery.Neural Regen Res. 2015;10(1):84-89.

[10] 谢利娜,刘艳辉,赵波,等.淫羊藿苷与BMP-2对体外培养成骨细胞增殖的影响[J].北京口腔医学,2018,26(6):319-322.

[11] LAI Y, CAO H, WANG X, et al. Porous composite scaffold incorporating osteogenic phytomolecule icariin for promoting skeletal regeneration in challenging osteonecrotic bone in rabbits. Biomaterials. 2018;153:1-13.

[12] 石永新,逄增金,羊明智,等. 3D打印技术制备聚磷酸钙/淫羊藿苷骨支架诱导骨髓间充质干细胞成骨分化治疗骨缺损[J].中国组织工程研究,2019,23(21):3309-3315.

[13] 管明强,朱志霞,周观明.淫羊藿苷对聚乳酸/纳米羟基磷灰石支架上成骨细胞增殖与分化的影响[J].包头医学院学报,2018,34(1): 92-94.

[14] 孔金海. PLLA/PLGA/HA复合材料的制备与评价[D].长沙:中南大学, 2009.

[15] RÓDENAS-ROCHINA J, RIBELLES JL, LEBOURG M. Comparative study of PCL-HAp and PCL-bioglass composite scaffolds for bone tissue engineering. J Mater Sci Mater Med. 2013;24(5):1293-1308.

[16] ROSETI L, PARISI V, PETRETTA M, et al. Scaffolds for Bone Tissue Engineering: State of the art and new perspectives. Mater Sci Eng C Mater Biol Appl. 2017;78:1246-1262.

[17] ZHAO W, LI J, JIN K, et al. Fabrication of functional PLGA-based electrospun scaffolds and their applications in biomedical engineering. Mater Sci Eng C Mater Biol Appl. 2016;59:1181-1194.

[18] DEL CASTILLO-SANTAELLA T, ORTEGA-OLLER I, PADIAL-MOLINA M, et al. Formulation, Colloidal Characterization, and In Vitro Biological Effect of BMP-2 Loaded PLGA Nanoparticles for Bone Regeneration. Pharmaceutics. 2019;11(8): E388.

[19] MARTINS C, SOUSA F, ARAÚJO F, et al. Functionalizing PLGA and PLGA Derivatives for Drug Delivery and Tissue Regeneration Applications. Adv Healthc Mater. 2018;7(1): UNSP 1701035.

[20] ZHAO X, HAN Y, LI J, et al. BMP-2 immobilized PLGA/hydroxyapatite fibrous scaffold via polydopamine stimulates osteoblast growth. Mater Sci Eng C Mater Biol Appl. 2017;78:658-666.

[21] HAIDER A, HAIDER S, HAN SS, et al. Recent advances in the synthesis, functionalization and biomedical applications of hydroxyapatite: a review. RSC Adv. 2017; 7(13): 7442-7458.

[22] ZHENG S, GUAN YH, YU HC, et al. Poly-L-lysine-coated PLGA/poly(amino acid)-modified hydroxyapatite porous scaffolds as efficient tissue engineering scaffolds for cell adhesion, proliferation, and differentiatio. New J Chem. 2019; 43(25): 9989-10002.

[23] 王帅,张霄雁,李哲海.骨髓间充质干细胞成骨细胞分化研究进展[J].医学综述,2015,21(12):2137-2139.

[24] XU H, GE YW, LU JW, et al. Icariin loaded-hollow bioglass/chitosan therapeutic scaffolds promote osteogenic differentiationand bone regeneration. Chem Eng J. 2018; 354: 285-294.

[25] WANG Z, WANG D, YANG D, et al. The effect of icariin on bone metabolism and its potential clinical application. Osteoporosis Int. 2018;29(3):535-544.

[26] FU N, DONG TZ, MENG A, et al. Research progress of the types and preparation techniques of scaffold materials in cartilage tissue engineering. Curr Stem Cell Res Ther. 2018; 13(7): 583-590.

[27] XIE YL, SUN WC, YAN FF, et al. Icariin-loaded porous scaffolds for bone regeneration through the regulation of the coupling process of osteogenesis and osteoclastic activity. Int J Nanomed. 2019; 14: 6019-6033.

[28] LUO Y, ZHANG YD, MIU GJ, et al. Icariin promotes osteoblast differentiation and inhibits adipogenesis of bone marrow stem cells through activation of Wnt/beta-catenin signaling in vivo and in vitro. Pharmazie.2018;73(8):459-464.

[29] ARIFFIN SHZ, MANOGARAN T, ABIDIN IZZ, et al. A perspective on stem cells as biological systems that produce differentiated osteoblasts and odontoblasts. Curr Stem Cell Res Ther. 2017; 12(3): 247-259.

[30] 郭晓宇. PI3K/AKT-eNOS介导淫羊藿苷促进骨髓基质细胞和成骨细胞的成骨性分化研究[D].甘肃:甘肃农业大学, 2013.