Reprogramming of mesenchymal stem cells derived from iPSCs seeded on biofunctionalized calcium phosphate scaffold for bone engineering

被引：95

作者：

Liu, Jun ^{[1
,2
]}

Chen, Wenchuan ^{[1
,2
]}

Zhao, Zhihe ^{[2
]}

Xu, Hockin H. K. ^{[1
,3
,4
,5
]}

机构：

[1] Univ Maryland, Sch Dent, Dept Endodont Prosthodont & Operat Dent, Biomat & Tissue Engn Div, Baltimore, MD 21201 USA

[2] Sichuan Univ, West China Hosp Stomatol, State Key Lab Oral Dis, Chengdu 610041, Peoples R China

[3] Univ Maryland, Sch Med, Ctr Stem Cell Biol & Regenerat Med, Baltimore, MD 21201 USA

[4] Univ Maryland, Sch Med, Marlene & Stewart Greenebaum Canc Ctr, Baltimore, MD 21201 USA

[5] Univ Maryland Baltimore Cty, Dept Mech Engn, Baltimore, MD 21250 USA

来源：

BIOMATERIALS | 2013年 / 34卷 / 32期

基金：

中国国家自然科学基金;

关键词：

Bone morphogenetic protein 2 (BMP2); Bone tissue engineering; Calcium phosphate cement (CPC); Gene transduction; Induced pluripotent stem cells (iPSCs); RGD immobilization; BMP-2; GENE-TRANSFER; MORPHOGENETIC PROTEIN-2; LENTIVIRAL VECTOR; UMBILICAL-CORD; IN-VITRO; HYDROXYAPATITE CEMENT; ALKALINE-PHOSPHATASE; FEMORAL DEFECTS; REGENERATION; EXPRESSION;

D O I：

10.1016/j.biomaterials.2013.07.029

中图分类号：

R318 [生物医学工程];

学科分类号：

0831 ;

摘要：

Human induced pluripotent stem cell-derived mesenchymal stem cells (iPSC-MSCs) are a promising choice of patient-specific stem cells with superior capability of cell expansion. There has been no report on bone morphogenic protein 2 (BMP2) gene modification of iPSC-MSCs for bone tissue engineering. The objectives of this study were to: (1) genetically modify iPSC-MSCs for BMP2 delivery; and (2) to seed BMP2 gene-modified iPSC-MSCs on calcium phosphate cement (CPC) immobilized with RGD for bone tissue engineering. iPSC-MSCs were infected with green fluorescence protein (GFP-iPSC-MSCs), or BMP2 lentivirus (BMP2-iPSC-MSCs). High levels of GFP expression were detected and more than 68% of GFP-iPSC-MSCs were GFP positive. BMP2-iPSC-MSCs expressed higher BMP2 levels than iPSC-MSCs in quantitative RT-PCR and ELISA assays (p < 0.05). BMP2-iPSC-MSCs did not compromise growth kinetics and cell cycle stages compared to iPSC-MSCs. After 14 d in osteogenic medium, ALP activity of BMP2-iPSC-MSCs was 1.8 times that of iPSC-MSCs (p < 0.05), indicating that BMP2 gene transduction of iPSC-MSCs enhanced osteogenic differentiation. BMP2-iPSC-MSCs were seeded on CPC scaffold bio-functionalized with RGD (RGD-CPC). BMP2-iPSC-MSC5 attached well on RGD-CPC. At 14 d, COL1A1 expression of BMP2-iPSC-MSCs was 1.9 times that of iPSC-MSCs. OC expression of BMP2-iPSC-MSCs was 2.3 times that of iPSC-MSCs. Bone matrix mineralization by BMP2-iPSC-MSCs was 1.8 times that of iPSC-MSCs at 21 d. In conclusion, iPSC-MSCs seeded on CPC were suitable for bone tissue engineering. BMP2 gene-modified iPSC-MSCs on RGD-CPC underwent osteogenic differentiation, and the overexpression of BMP2 in iPSC-MSCs enhanced differentiation and bone mineral production on RGD-CPC. BMP2-iPSC-MSC seeding on RGD-CPC scaffold is promising to enhance bone regeneration efficacy. (C) 2013 Elsevier Ltd. All rights reserved.

引用

页码：7862 / 7872

页数：11

共 66 条

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