Control of adhesive ligand density for modulation of nucleus pulposus cell phenotype

Barcellona, Marcos N.; Speer, Julie E.; Fearing, Bailey V.; Jing, Liufang; Pathak, Amit; Gupta, Munish C.; Buchowski, Jacob M.; Kelly, Michael P.; Setton, Lori A.

doi:10.1016/j.biomaterials.2020.120057

Cited by 31 publications

(65 citation statements)

References 49 publications

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“…However, over confluence of cells also may decrease cell viability. Additionally, the circularity of NPCs with PPARδ agonist treatment showed a more "naïve" morphology, as compared to the control [29].…”

Section: Discussionmentioning

confidence: 89%

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Towards Tissue-Specific Stem Cell Therapy for the Intervertebral Disc: PPARδ Agonist Increases the Yield of Human Nucleus Pulposus Progenitor Cells in Expansion

et al. 2021

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(1) Background: Low back pain (LBP) is often associated with intervertebral disc degeneration (IVDD). Autochthonous progenitor cells isolated from the center, i.e., the nucleus pulposus, of the IVD (so-called nucleus pulposus progenitor cells (NPPCs)) could be a future cell source for therapy. The NPPCs were also identified to be positive for the angiopoietin-1 receptor (Tie2). Similar to hematopoietic stem cells, Tie2 might be involved in peroxisome proliferator-activated receptor delta (PPARδ) agonist-induced self-renewal regulation. The purpose of this study was to investigate whether a PPARδ agonist (GW501516) increases the Tie2+ NPPCs’ yield within the heterogeneous nucleus pulposus cell (NPC) population. (2) Methods: Primary NPCs were treated with 10 µM of GW501516 for eight days. Mitochondrial mass was determined by microscopy, using mitotracker red dye, and the relative gene expression was quantified by qPCR, using extracellular matrix and mitophagy-related genes. (3) The NPC’s group treated with the PPARδ agonist showed a significant increase of the Tie2+ NPCs yield from ~7% in passage 1 to ~50% in passage two, compared to the NPCs vehicle-treated group. Furthermore, no significant differences were found among treatment and control, using qPCR and mitotracker deep red. (4) Conclusion: PPARδ agonist could help to increase the Tie2+ NPCs yield during NPC expansion.

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Section: Discussionmentioning

confidence: 89%

“…Cells' morphology was quantified by using ImageJ v1.53c software on the microscopic pictures, using "cell body circularity" functionality [28,29]. An example of how the cells' outlines were defined by manual segmentation can be found in a previous study [17].…”

Section: Visualisation Of Cell's Morphologymentioning

confidence: 99%

Towards Tissue-Specific Stem Cell Therapy for the Intervertebral Disc: PPARδ Agonist Increases the Yield of Human Nucleus Pulposus Progenitor Cells in Expansion

et al. 2021

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“…To better visualize the morphology of cells, live cells were pre-stained with 2μM Calcein-AM for 10 min at 37 °C. Cell’s morphology was then quantified by ImageJ v1.53c on the microscopic pictures using “cell body circularity” [ 33 , 46 ]. An example of how the cells outlines were defined by manual segmentation can be found in the Supplementary Materials file S2 .…”

Section: Methodsmentioning

confidence: 99%

Spheroid-Like Cultures for Expanding Angiopoietin Receptor-1 (aka. Tie2) Positive Cells from the Human Intervertebral Disc

Zhang

Guerrero

Croft

et al. 2020

IJMS

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Lower back pain is a leading cause of disability worldwide. The recovery of nucleus pulposus (NP) progenitor cells (NPPCs) from the intervertebral disc (IVD) holds high promise for future cell therapy. NPPCs are positive for the angiopoietin-1 receptor (Tie2) and possess stemness capacity. However, the limited Tie2+ NPC yield has been a challenge for their use in cell-based therapy for regenerative medicine. In this study, we attempted to expand NPPCs from the whole NP cell population by spheroid-formation assay. Flow cytometry was used to quantify the percentage of NPPCs with Tie2-antibody in human primary NP cells (NPCs). Cell proliferation was assessed using the population doublings level (PDL) measurement. Synthesis and presence of extracellular matrix (ECM) from NPC spheroids were confirmed by quantitative Polymerase Chain Reaction (qPCR), immunostaining, and microscopy. Compared with monolayer, the spheroid-formation assay enriched the percentage of Tie2+ in NPCs’ population from ~10% to ~36%. Moreover, the spheroid-formation assay also inhibited the proliferation of the Tie2- NPCs with nearly no PDL. After one additional passage (P) using the spheroid-formation assay, NPC spheroids presented a Tie2+ percentage even further by ~10% in the NPC population. Our study concludes that the use of a spheroid culture system could be successfully applied to the culture and expansion of tissue-specific progenitors.

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“…Changes have also been observed in the presence of matrix degrading enzymes such as the MMP and ADAMTS families, which results in an altered cellular microenvironment and a decreased ability for tissue repair and homeostasis 4,[15][16][17] . As the NP has limited ability for self-repair, biomaterials have been investigated as a means to deliver cells to the degenerative disc or to provide stimuli to modulate cell (endogenous or exogenous) behavior and promote IVD regeneration 14,18,19 .…”

Section: Introductionmentioning

confidence: 99%

“…In addition to ligand presentation, other biomaterial parameters including substrate stiffness, porosity, topography, and surface chemistry have been tested for their ability to modulate cell behaviors 27-33 . 14,20,34-37 Previous studies have shown that culture of NP cells upon soft (~0.5 kPa) hydrogel substrates functionalized with laminin-111 can be used to promote the cell behaviors and gene expression profiles characteristically observed in the juvenile NP, while cells cultured on stiff (10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20) hydrogel substrates or tissue culture plastic exhibit a fibroblast-like phenotype 9,38,39 . While this work made use of full-length laminin proteins, short peptide sequences offers several advantages in biomaterial design including the opportunity to precisely control sites of biomaterial and cell interactions 24,39 .…”

Section: Introductionmentioning

confidence: 99%

Engineered Peptide-Functionalized Hydrogels Modulate the RNA Transcriptome of Human Nucleus Pulposus Cells In Vitro

Barcellona

Speer

Jing

et al. 2021

Preprint

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Degeneration and aging of the nucleus pulposus (NP) of the intervertebral disc (IVD) is accompanied by alterations in NP cell phenotype marked by a shift towards a fibroblast-like, catabolic state. We have recently demonstrated an ability to manipulate the phenotype of human adult degenerative NP cells through 2D culture upon poly(ethylene glycol) (PEG) based hydrogels dually functionalized with integrin- and syndecan-binding laminin-mimetic peptides (LMPs). In the present study, we sought to understand the transcriptomic changes elicited through NP cell interactions with the LMP-functionalized hydrogel system (LMP gel) by examining targets of interest a priori and by conducting unbiased analysis to identify novel mechanosensitive targets. The results of gene specific analysis demonstrated that the LMP gel promoted adult degenerative NP cells to upregulate 148 genes including several NP markers (e.g. NOG and ITGA6) and downregulate 277 genes, namely several known fibroblastic markers. Additionally, 13 genes associated with G protein-coupled receptors, many of which are known drug targets, were identified as differentially regulated following culture upon the gel. Furthermore, through gene set enrichment analysis we identified over 700 pathways enriched amongst the up- and downregulated genes including pathways related to cell differentiation, notochord morphogenesis, and intracellular signaling. Together these findings demonstrate the global mechanobiological effects induced by the LMP gel and confirm the ability of this substrate to modulate NP cell phenotype.

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Control of adhesive ligand density for modulation of nucleus pulposus cell phenotype

Cited by 31 publications

References 49 publications

Towards Tissue-Specific Stem Cell Therapy for the Intervertebral Disc: PPARδ Agonist Increases the Yield of Human Nucleus Pulposus Progenitor Cells in Expansion

Towards Tissue-Specific Stem Cell Therapy for the Intervertebral Disc: PPARδ Agonist Increases the Yield of Human Nucleus Pulposus Progenitor Cells in Expansion

Spheroid-Like Cultures for Expanding Angiopoietin Receptor-1 (aka. Tie2) Positive Cells from the Human Intervertebral Disc

Engineered Peptide-Functionalized Hydrogels Modulate the RNA Transcriptome of Human Nucleus Pulposus Cells In Vitro

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