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基因调控纳米颗粒复合物与 pDNA 和 shRNA 的转染可控制 hMSCs 的多谱系分化。

Transfection of gene regulation nanoparticles complexed with pDNA and shRNA controls multilineage differentiation of hMSCs.

机构信息

Department of Nano-regenerative Medical Engineering, College of Life Science, CHA University, 6F, CHA Biocomplex, Sampyeong-Dong, Bundang-gu, Seongnam-si, 13488, Republic of Korea.

Department of Nano-regenerative Medical Engineering, College of Life Science, CHA University, 6F, CHA Biocomplex, Sampyeong-Dong, Bundang-gu, Seongnam-si, 13488, Republic of Korea.

出版信息

Biomaterials. 2018 Sep;177:1-13. doi: 10.1016/j.biomaterials.2018.05.035. Epub 2018 May 29.

DOI:10.1016/j.biomaterials.2018.05.035
PMID:29883913
Abstract

Overexpression and knockdown of specific proteins can control stem cell differentiation for therapeutic purposes. In this study, we fabricated RUNX2, SOX9, and C/EBPα plasmid DNAs (pDNAs) and ATF4-targeting shRNA (shATF4) to induce osteogenesis, chondrogenesis, and adipogenesis of human mesenchymal stem cells (hMSCs). The pDNAs and shATF4 were complexed with TRITC-gene regulation nanoparticles (GRN). Osteogenesis-related gene expression was reduced at early (12 h) and late (36 h) time points after co-delivery of shATF4 and SOX9 or C/EBPα pDNA, respectively, and osteogenesis was inhibited in these hMSCs. By contrast, osteogenesis-related genes were highly expressed upon co-delivery of RUNX2 and ATF4 pDNAs. DEX in GRN enhanced chondrogenic differentiation. Expression of osteogenesis-, chondrogenesis-, and adipogenesis-related genes was higher in hMSCs transfected with NPs complexed with RUNX2 and ATF4 pDNAs, shATF4 and SOX9 pDNA, and shATF4 and C/EBPα pDNA for 72 h than in control hMSCs, respectively. Moreover, delivery of these NPs also increased expression of osteogenesis-, chondrogenesis-, and adipogenesis-related proteins. These alterations in expression led to morphological changes, indicating that hMSCs differentiated into osteoblasts, chondrocytes, and adipose cells.

摘要

过表达和敲低特定蛋白质可以控制干细胞分化，以达到治疗目的。在这项研究中，我们构建了 RUNX2、SOX9 和 C/EBPα 质粒 DNA（pDNA）和 ATF4 靶向 shRNA（shATF4），以诱导人间充质干细胞（hMSC）的成骨、成软骨和成脂分化。pDNA 和 shATF4 与 TRITC-基因调控纳米颗粒（GRN）复合。在共递送 shATF4 和 SOX9 或 C/EBPα pDNA 后，分别在早期（12 h）和晚期（36 h）时间点，成骨相关基因表达降低，这些 hMSC 中的成骨作用受到抑制。相比之下，共递送 RUNX2 和 ATF4 pDNA 时，成骨相关基因高度表达。DEX 在 GRN 中增强了软骨分化。在转染了与 RUNX2 和 ATF4 pDNA、shATF4 和 SOX9 pDNA 以及 shATF4 和 C/EBPα pDNA 复合的 NPs 的 hMSC 中，成骨、成软骨和成脂相关基因的表达在 72 h 时高于对照组 hMSC，分别。此外，这些 NPs 的递送还增加了成骨、成软骨和成脂相关蛋白的表达。这些表达的改变导致形态变化，表明 hMSC 分化为成骨细胞、软骨细胞和脂肪细胞。

相似文献

[1]
Transfection of gene regulation nanoparticles complexed with pDNA and shRNA controls multilineage differentiation of hMSCs.

Biomaterials. 2018-5-29

[2]
Co-delivery of SOX9 genes and anti-Cbfa-1 siRNA coated onto PLGA nanoparticles for chondrogenesis of human MSCs.

Biomaterials. 2012-3-15

[3]
Sequential transfection of RUNX2/SP7 and ATF4 coated onto dexamethasone-loaded nanospheresenhances osteogenesis.

Sci Rep. 2018-1-23

[4]
The use of biodegradable PLGA nanoparticles to mediate SOX9 gene delivery in human mesenchymal stem cells (hMSCs) and induce chondrogenesis.

Biomaterials. 2010-9-27

[5]
Multiply clustered gold-based nanoparticles complexed with exogenous pDNA achieve prolonged gene expression in stem cells.

Theranostics. 2019-7-9

[6]
Co-Delivery of RUNX2-Targeting miRNAs and shRNAs Using Nanoparticles Composed of Dexamethasone and PEI Induces Chondrogenesis of Human Mesenchymal Stem Cells.

J Biomed Nanotechnol. 2019-1-1

[7]
Co-delivery of Cbfa-1-targeting siRNA and SOX9 protein using PLGA nanoparticles to induce chondrogenesis of human mesenchymal stem cells.

Biomaterials. 2014-6-23

[8]
Gene expression profiling of chondrogenic differentiation by dexamethasone-conjugated polyethyleneimine with SOX trio genes in stem cells.

Stem Cell Res Ther. 2018-12-7

[9]
SOX9 gene transfer via safe, stable, replication-defective recombinant adeno-associated virus vectors as a novel, powerful tool to enhance the chondrogenic potential of human mesenchymal stem cells.

Stem Cell Res Ther. 2012

[10]
Multilineage differentiation of human-derived dermal fibroblasts transfected with genes coated on PLGA nanoparticles plus growth factors.

Biomaterials. 2012-10-22

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Pharmaceutics. 2024-8-16

[2]
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World J Stem Cells. 2023-6-26

[3]
PSAT1 positively regulates the osteogenic lineage differentiation of periodontal ligament stem cells through the ATF4/PSAT1/Akt/GSK3β/β-catenin axis.

J Transl Med. 2023-2-2

[4]
Biomaterial-assisted biotherapy: A brief review of biomaterials used in drug delivery, vaccine development, gene therapy, and stem cell therapy.

Bioact Mater. 2022-1-19

[5]
Recent Developed Strategies for Enhancing Chondrogenic Differentiation of MSC: Impact on MSC-Based Therapy for Cartilage Regeneration.

Stem Cells Int. 2021-3-20

[6]
Circular RNA SIPA1L1 promotes osteogenesis via regulating the miR-617/Smad3 axis in dental pulp stem cells.

Stem Cell Res Ther. 2020-8-24

[7]
Human amnion-derived mesenchymal stem cells promote osteogenic differentiation of human bone marrow mesenchymal stem cells via H19/miR-675/APC axis.

Aging (Albany NY). 2020-5-20

[8]
LncRNA H19 promotes the committed differentiation of stem cells from apical papilla via miR-141/SPAG9 pathway.

Cell Death Dis. 2019-2-12

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