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Decoupling the effects of nanopore size and surface roughness on the attachment, spreading and differentiation of bone marrow-derived stem cells.

ABSTRACT: The nanopore size and roughness of nanoporous surface are two critical variables in determining stem cell fate, but little is known about the contribution from each cue individually. To address this gap, we use two-dimensional nanoporous membranes with controlled nanopore size and roughness to culture bone marrow-derived mesenchymal stem cells (BMSCs), and study their behaviors such as attachment, spreading and differentiation. We find that increasing the roughness of nanoporous surface has no noticeable effect on cell attachment, and only slightly decreases cell spreading areas and inhibits osteogenic differentiation. However, BMSCs cultured on membranes with larger nanopores have significantly fewer attached cells and larger spreading areas. Moreover, these cells cultured on larger nanopores undergo enhanced osteogenic differentiation by expressing more alkaline phosphatase, osteocalcin, osteopontin, and secreting more collagen type I. These results suggest that although both nanopore size and roughness can affect BMSCs, nanopore size plays a more significant role than roughness in controlling BMSC behavior.

SUBMITTER: Xia J

PROVIDER: S-EPMC7262959 | biostudies-literature | 2020 Jul

REPOSITORIES: biostudies-literature

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Decoupling the effects of nanopore size and surface roughness on the attachment, spreading and differentiation of bone marrow-derived stem cells.

Xia Jing J Yuan Yuan Y Wu Huayin H Huang Yuting Y Weitz David A DA

Biomaterials 20200331

The nanopore size and roughness of nanoporous surface are two critical variables in determining stem cell fate, but little is known about the contribution from each cue individually. To address this gap, we use two-dimensional nanoporous membranes with controlled nanopore size and roughness to culture bone marrow-derived mesenchymal stem cells (BMSCs), and study their behaviors such as attachment, spreading and differentiation. We find that increasing the roughness of nanoporous surface has no n ...[more]

PMID: 32276040

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Project description:BackgroundSystemic inflammation is the main factor underlying secondary osteoporosis in patients with rheumatoid arthritis (RA). Janus kinase inhibitors (JAKi), such as tofacitinib (Tofa), can control systemic inflammation and may have beneficial effects on bone in various models. This might be due to direct effects on the bone microenvironment and not exclusively based on their anti-inflammatory function. Bone marrow adipocytes (BMAds) are abundant in the bone microenvironment. The effect of JAKi on BMAds is unknown, but evidence suggests that there is competition between human bone marrow-derived stromal cell (hBMSC) differentiation routes towards BMAds and osteoblasts (Ob) in osteoporosis.ObjectivesThe aims of the study are to determine whether Tofa influences BMAds and Ob derived from hBMSCs and to investigate the potential effects of Tofa on bone marrow adiposity in RA patients.MethodsTo determine the effect of Tofa on cellular commitment, hBMSCs were differentiated to BMAds or OBs for 3 days together with Tofa at 200, 400, or 800 nM and TNFα. This study was also conducted using differentiated BMAds. The impact of Tofa was determined by gene and protein expression analysis and cell density monitoring. In parallel, in a pilot study of 9 RA patients treated with Tofa 5 mg twice a day (NCT04175886), the proton density fat fraction (PDFF) was measured using MRI at the lumbar spine at baseline and at 6 months.ResultsIn non-inflammatory conditions, the gene expression of Runx2 and Dlx5 decreased in Ob treated with Tofa (p <0.05). The gene expression of PPARγ2, C/EBPα, and Perilipin 1 were increased compared to controls (p <0.05) in BMAds treated with Tofa. Under inflammatory conditions, Tofa did not change the expression profiles of Ob compared to TNFα controls. In contrast, Tofa limited the negative effect of TNFα on BMAd differentiation (p <0.05). An increase in the density of differentiated BMAds treated with Tofa under TNFα was noted (p <0.001). These findings were consolidated by an increase in PDFF at 6 months of treatment with Tofa in RA patients (46.3 ± 7.0% versus 53.2 ± 9.2% p <0.01).ConclusionTogether, these results suggest a stimulatory effect of Tofa on BMAd commitment and differentiation, which does not support a positive effect of Tofa on bone.

Dataset Information

Decoupling the effects of nanopore size and surface roughness on the attachment, spreading and differentiation of bone marrow-derived stem cells.

Publications

Decoupling the effects of nanopore size and surface roughness on the attachment, spreading and differentiation of bone marrow-derived stem cells.

Similar Datasets

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