Transfection of the IHH gene into rabbit BMSCs in a simulated microgravity environment promotes chondrogenic differentiation and inhibits cartilage aging.

Peng-Cheng Liu,Jun-Feng Liu,Li-Yang Chen,Kuan Liu,Xing Wu,Kuo Xia

doi:10.18632/oncotarget.11871

Peng-Cheng Liu, Jun-Feng Liu + Show 4 more

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https://doi.org/10.18632/oncotarget.11871

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Abstract

The effect of overexpressing the Indian hedgehog (IHH) gene on the chondrogenic differentiation of rabbit bone marrow-derived mesenchymal stem cells (BMSCs) was investigated in a simulated microgravity environment. An adenovirus plasmid encoding the rabbit IHH gene was constructed in vitro and transfected into rabbit BMSCs. Two large groups were used: conventional cell culture and induction model group and simulated microgravity environment group. Each large group was further divided into blank control group, GFP transfection group, and IHH transfection group. During differentiation induction, the expression levels of cartilage-related and cartilage hypertrophy-related genes and proteins in each group were determined. In the conventional model, the IHH transfection group expressed high levels of cartilage-related factors (Coll2 and ANCN) at the early stage of differentiation induction and expressed high levels of cartilage hypertrophy-related factors (Coll10, annexin 5, and ALP) at the late stage. Under the simulated microgravity environment, the IHH transfection group expressed high levels of cartilage-related factors and low levels of cartilage hypertrophy-related factors at all stages of differentiation induction. Under the simulated microgravity environment, transfection of the IHH gene into BMSCs effectively promoted the generation of cartilage and inhibited cartilage aging and osteogenesis. Therefore, this technique is suitable for cartilage tissue engineering.

Highlights

Cartilage tissue engineering provides new ideas for the treatment of cartilage damage and defects
Studies showed that transfection of Indian hedgehog (IHH) into Bone marrow-derived mesenchymal stem cells (BMSCs), either alone or in combination with bone morphogenetic protein-2 (BMP-2) or transforming growth www.impactjournals.com/oncotarget factor-β (TGF-β), promoted chondrogenesis in primary BMSCs [8], which provided the theoretical basis for using IHH gene transfection to construct tissue-engineered cartilage
Chondrogenesis in BMSCs is regulated by a variety of signaling pathways acting together

Summary

Introduction

Cartilage tissue engineering provides new ideas for the treatment of cartilage damage and defects. Indian hedgehog (IHH) is a conserved protein in the hedgehog signaling pathway and performs important regulatory functions in cartilage hypertrophy and bone formation. Defects in the IHH gene or protein result in the malformation of extremities, along with both a reduction in chondrocyte proliferation and swelling of hypertrophic chondrocytes [6]. IHH is currently thought to promote increased parathyroid hormone/parathyroid-hormone-related peptide (PTH/PTHrP) expression in chondrocytes surrounding joints; the latter can interact with its receptor, parathyroid hormone receptor type 1 (Pthr-1), to promote the proliferation and inhibit the hypertrophy of chondrocytes. Studies showed that transfection of IHH into BMSCs, either alone or in combination with bone morphogenetic protein-2 (BMP-2) or transforming growth www.impactjournals.com/oncotarget factor-β (TGF-β), promoted chondrogenesis in primary BMSCs [8], which provided the theoretical basis for using IHH gene transfection to construct tissue-engineered cartilage

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