Inflammation-mediated age-dependent effects of casein kinase 2-interacting protein-1 on osteogenesis in mesenchymal stem cells

Abstract

Background The casein kinase 2-interacting protein-1 ( CKIP-1 ) is important in the development of osteoblasts and cardiomyocytes. However, the effects of CKIP-1 on osteoblast precursor mesenchymal stem cells (MSCs) remain unclear. This study aimed to determine whether CKIP-1 affects osteogenic differentiation in MSCs and explore the relationship of CKIP-1 and inflammation. Methods Bone marrow MSCs of CKIP-1 wild type (WT) and knockout (KO) mice were cultivated in vitro . Cell phenotype was analyzed by flow cytometry, colony formation was detected to study the proliferative ability. Osteogenic and adipogenic induction were performed. The osteogenic ability was explored by alizarin red staining, alkaline phosphatase (ALP) staining and ALP activity detection. Quantitative real-time polymerase chain reaction (qRT-PCR) was carried out to determine the mRNA expression levels of osteoblast marker genes. The adipogenic ability was detected by oil red O staining. Content of the bone was analyzed to observe the differences of bone imaging parameters including trabecular bone volume/tissue volume (BV/TV), bone surface area fraction/trabecular BV, trabecular number (Tb.N), and trabecular spacing (Tb.sp). Interleukin (IL)-1β was injected on WT mice of 2 months old and 18 months old, respectively. Difference in CKIP-1 expression was detected by RT-PCR and western blot. The relationship between CKIP-1 and inflammation was explored by RT-PCR and western blot. Results ALP assays, alizarin red staining, and qRT-PCR showed that MSCs derived from CKIP-1 KO mice exhibited a stronger capability for osteogenesis. Micro-computed tomography detection showed that among 18-month-old mice, CKIP-1 KO mice presented significantly higher bone mass compared with WT mice ( P = 0.02). No significant difference was observed in 2-month-old mice. In vivo data showed that expression of CKIP-1 was higher in the bone marrow of aging mice than in young mice (4.3-fold increase at the mRNA level, P = 0.04). Finally, the expression levels of CKIP-1 in bone marrow (3.2-fold increase at the mRNA level, P = 0.03) and cultured MSCs were up-regulated on chronic inflammatory stimulation by IL-1β. Conclusions CKIP-1 is responsible for negative regulation of MSC osteogenesis with age-dependent effects. Increasing levels of inflammation with aging may be the primary factor responsible for higher expression levels of CKIP-1 but may not necessarily affect MSC aging.

Ckip-1 Mediates P. gingivalis–Suppressed Cementoblast Mineralization

“…It is worth mentioning that Ckip-1 may also play a regulatory role in inflammation. As proved in most of the studies, Ckip-1 was positively correlated with the inflammation of various cells, such as aging-mediated inflammation of bone marrow mesenchymal stem cells (Tian et al 2020), proinflammatory polarization of macrophages (Chen et al 2017), and LPS-triggered inflammation of C3H/10T1/2 cells (He et al 2019). However, 1 reverse case in which Ckip-1 overexpression could inhibit high glucose–induced inflammation in human retinal endothelial cells was also reported (Zhang et al 2019).…”

Corrigendum: Inflammation-mediated age-dependent effects of casein kinase 2-interacting protein-1 on osteogenesis in mesenchymal stem cells

“…Following the original article's publication, [1] the authors informed us of the following errors: [1] The Figure 1 was submitted incorrectly, while correct Figure 1 should be submitted as following:…”