The effect of high glucose on the biological characteristics of nucleus pulposus‐derived mesenchymal stem cells

Abstract

Diabetes mellitus (DM) is a dependent risk factor in the progression of intervertebral disc degeneration (IVDD). High glucose supply has negative effects on nucleus pulpous (NP) cell and mesenchymal stem cell (MSC) biology. However, the effect of hyperglycaemia on the biological characterization of nucleus pulpous-derived mesenchymal stem cell (NPMSC) has not been investigated previously. Therefore, further exploration of the effects of DM-associated hyperglycaemia on NPMSC biology is important to better understand and develop endogenous repair strategies of DM patient-associated IVDD. Therefore, the cell biological characteristics were compared between NPMSC cultured in media with low glucose concentration (LG-NPMSC) and high glucose concentration (HG-NPMSC). The results demonstrated that HG-NPMSC showed significantly decreased cell proliferation, colony formation ability, migration and wound-healing capability compared with those of LG-NPMSC. HG-NPMSC also showed significantly decreased expressions of stemness genes and mRNA and protein expressions of silent information regulator protein 1 (SIRT1), SIRT6, hypoxia inducible factor-1α (HIF-1α) and glucose transporter 1 (GLUT-1), whereas increased cell apoptosis, cell senescence and caspase-3 expression. These results suggest that high glucose may decrease proliferation and stemness maintenance ability and increase apoptosis and senescence of NPMSC.Significance of the study: We found that high glucose concentration significantly decreased cell proliferation, colony formation ability, migration and wound-healing capability of nucleus pulposus-derived mesenchymal stem cells. Moreover, high glucose cultured nucleus pulposus-derived mesenchymal stem cells showed significantly decreased expression of stemness genes, related mRNA and protein, whereas increased cell apoptosis, cell senescence and expression of caspase-3. The present study indicated that better control of high concentration glucose in the early stage of diabetes mellitus should be recommended to prevent or limit intervertebral disc degeneration. K E Y W O R D S-high glucose, nucleus pulposus-derived mesenchymal stem cells, proliferation, apoptosis, senescence

RACK1 mediates the advanced glycation end product‐induced degradation of HIF‐1α in nucleus pulposus cells via competing with HSP90 for HIF‐1α binding

“…Previous work has clearly demonstrated that diabetes is associated with more rapid IVDD progression (X. Liu et al, 2018). Hyperglycemia‐induced AGE accumulation within the IVD has also been shown to elevate the risk of NPC dysfunction (Y. Liu et al, 2020; Tsai et al, 2014), and diabetic NPCs have been found to exhibit poorer survival and enhanced ECM degradation following AGE exposure (Song et al, 2017; Tsai et al, 2014). The specific mechanisms whereby AGEs modulate NPC functionality, however, have not been well‐defined in these previous studies.…”

Comparison and Optimization: Different Medium and a Novel Scheme for Rabbit Bone Marrow Mesenchymal Stem Cells Culture

“…Studies have shown that medium with high glucose concentration within a certain range could effectively improve the proliferation ability of MSCs [23,24]. However, some studies also have shown that medium with high glucose might accelerate cell senescence and apoptosis in seveal pathways [25][26][27]. Yanan Kong et al cultured MSCs with high glucose (33 mmol/L) medium, and found that the apoptosis rate of cells remained low after culturing for 1 day, while the apoptosis rate of cells increased greatly after culturing for 5 days [25].…”

“…Tzu-Ching C et al cultured BMSCs with low glucose (5.5 mmol/L) and high glucose (25 mmol/L) medium respectively, they also found that high glucose could accelerate the senescence of BMSCs by promoting their ability of autophagy after culturing for 2 weeks [26]. Yang Liu et al cultured nucleus pulposus derived MSCs (NPMSCs) with low glucose (5.5 mmol/L) and high glucose (25 mmol/L) medium respectively, and they found that high glucose medium cultured NPMSCs showed signi cantly decreased expression levels of stemness genes, related mRNA and protein, whereas increased expression levels of cell senescence markers and caspase-3 [27]. Cramer et al applied medium with different concentration glucose (5.56mmol/L, 13.9mmol/L, 27.8mmol/L and 55.6mmol/L) to culture adipose derived stem cells (ADSCs) for 120 hours, and found that the hyperglycemia microenvironment could simultaneously reduce the proliferation ability of ADSCs and increase their apoptosis [28].…”

Towards Physiologic Culture Approaches to Improve Standard Cultivation of Mesenchymal Stem Cells

“…Most studies under normoxic culture conditions support that high glucose concentration (5000 mg/L) in the culture media can suppress proliferation of bone marrow [ 46 , 47 , 48 ], Bichat’s buccal fat pad [ 49 ] and nucleus pulposus MSCs [ 50 ], reduce their colony forming ability [ 50 ], induce cellular senescence [ 47 , 50 , 51 ], alter MSC spindle-shape morphology [ 47 ], and upregulate autophagy [ 50 , 51 ], compared with low glucose (1000 mg/L) media concentration. In addition, MSCs cultured in low glucose media have been shown to maintain their differentiation capacities and stemness [ 46 , 47 , 50 ], and increase antioxidant enzyme expression and mitochondrial respiration [ 47 ]. In contrast with these results, a few studies have reported that high glucose does not have an effect on the apoptosis and proliferation rates of adipose-derived [ 52 ] and primary BM-MSCs [ 53 ], while it can significantly enhance proliferation of telomerase-immortalized MSCs.…”