Pyrophosphate Stimulates Differentiation, Matrix Gene Expression and Alkaline Phosphatase Activity in Osteoblasts

Pujari-Palmer, Michael; Pujari-Palmer, Shiuli; Lu, Xunyu; Lind, Thomas; Melhus, Håkan; Engstrand, Thomas; Karlsson-Ott, Marjam; Engqvist, Håkan

doi:10.1371/journal.pone.0163530

Cited by 48 publications

(56 citation statements)

References 49 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…1C). We then examined the early osteogenic markers ALP [17,62,63] and mineral node formation. ALP activity was analyzed by histochemical staining.…”

Section: Resultsmentioning

confidence: 99%

Bone morphogenetic protein 9 induces osteogenic differentiation of germ cell 1 spermatogonial cells

Zhang

Xu²,

Chen³

et al. 2019

Preprint

View full text Add to dashboard Cite

30Germ cell 1 spermatogonial (GC-1spg) cells are multipotent progenitor cells. We 31 previously confirmed that bone morphogenetic protein (BMP) 9 is among the most 32 osteogenic BMPs. However, whether GC-1spg cells are driven toward osteogenic 33 differentiation under proper stimuli is uncertain. Additionally, the molecular 34 mechanism of BMP9 remains unclear. In the present study, we aimed to determine 35 whether BMP9 can induce osteogenic differentiation of GC-1spg cells. Recombinant 36 adenoviruses were generated by the AdEasy system to regulate the BMP9 expression in 37 GC-1spg cells. We identified osteogenic markers by real-time PCR and staining 38 techniques in vitro. Ectopic ossification assays and histological analysis were also 39 performed to verify the in vivo activity of BMP9. Finally, potential signaling pathways 40 of BMP9 were assessed by transcriptome sequencing and KEGG enrichment analysis. 41Using recombinant adenoviruses, we demonstrate that BMP9 upregulates osteogenic 42 markers including Runx2, osteocalcin, osteopontin, and Sox9. BMP9 also activates 43 alkaline phosphatase activity and calcium deposition in GC-1spg cells. In vivo results 44show that BMP9 overexpression in GC-1spg cells promotes ectopic bone formation and 45 chondrogenesis. In addition, RNA-sequencing and KEGG pathway analysis 3 46 demonstrate that several signaling pathways are involved in BMP9-mediated 47 osteogenesis. GC-1spg cells not only maintain spermatogenesis but also retain the 48 ability to form bone tissue. Therefore, BMP9 activity in GC-1spg cells may help 49 identify signaling pathways implicated in bone formation and could be of use in 50 regenerative medicine. 51 Keywords: Bone morphogenetic protein 9, RNA-seq, KEGG pathway analysis, 52 recombinant adenovirus, chondrogenesis 53 Introduction 54 Germ cell 1 spermatogonial (GC-1spg) cells are mouse spermatogonia that are 55 immortalized by the SV40 large T antigen [1]. These cells can differentiate into diverse cell 56 types or undergo self-renewal, depending on different factors [2,3,4]. GC-1spg cells are 57 largely involved in spermatogenesis and have mesenchymal stem cell (MSC) differentiation 58 potential [5,6,7,8]. In other words, GC-1spg cells can differentiate into different cell types, 59including those of the osteogenic, chondrogenic, or adipogenic lineages [9,10,11,12]. 60However, several molecular events [13,14] and factors are involved in osteogenic 61 differentiation of MSCs. Importantly, bone morphogenetic proteins (BMPs) are essential for 62 this process [15,16,17]. 63 BMPs, members of the TGF-β superfamily, are required for bone formation, stem cell 64 differentiation, and male reproduction [15,16,18,19,20]. Thus far, more than 15 different 65 BMPs have been identified. Our previous work showed that BMP9 is the most osteogenic 66 BMP [21,22,23,24]. BMP9, also known as growth differentiation factor 2 (GDF-2)[25], was 67 originally isolated from the fetal mouse liver [25,26]. Bone formation involves BMP9 acting 68 through the Notch signaling ...

show abstract

“…1C). We then examined the early osteogenic markers ALP [17,62,63] and mineral node formation. ALP activity was analyzed by histochemical staining.…”

Section: Resultsmentioning

confidence: 99%

Bone morphogenetic protein 9 induces osteogenic differentiation of germ cell 1 spermatogonial cells

Zhang

Xu²,

Chen³

et al. 2019

Preprint

View full text Add to dashboard Cite

show abstract

“…Interestingly, a recent study has shown that high extracellular Pi induced phenotypic transformation of vascular smooth muscle cells into osteogenic-like cells via the activation of Wnt/β-catenin signaling in these cells [42]. Our previous study, however, also suggested that extracellular PPi directly stimulates osteogenesis [2, 43]. Therefore, it is possible that extracellular PPi resulting from ANK transport directly or via its hydrolysis to extracellular Pi may affect Wnt/β-catenin signaling in osteogenic precursor cells.…”

Section: Discussionmentioning

confidence: 98%

The role of the progressive ankylosis protein (ANK) in adipogenic/osteogenic fate decision of precursor cells

Minashima

Quirno

Lee

et al. 2017

Bone

View full text Add to dashboard Cite

The progressive ankylosis protein (ANK) is a transmembrane protein that transports intracellular pyrophosphate (PPi) to the extracellular milieu. In this study we show increased fatty degeneration of the bone marrow of adult ank/ank mice, which lack a functional ANK protein. In addition, isolated bone marrow stromal cells (BMSCs) isolated from ank/ank mice showed a decreased proliferation rate and osteogenic differentiation potential, and an increased adipogenic differentiation potential compared to BMSCs isolated from wild type (WT) littermates. Wnt signaling pathway PCR array analysis revealed that Wnt ligands, Wnt receptors and Wnt signaling proteins that stimulate osteoblast differentiation were expressed at markedly lower levels in ank/ank BMSCs than in WT BMSCs. Lack of ANK function also resulted in impaired bone fracture healing, as indicated by a smaller callus formed and delayed bone formation in the callus site. Whereas 5 weeks after fracture, the fractured bone in WT mice was further remodeled and restored to original shape, the fractured bone in ank/ank mice was not fully restored and remodeled to original shape. In conclusion, our study provides evidence that ANK plays a critical role in the adipogenic/osteogenic fate decision of adult mesenchymal precursor cells. ANK functions in precursor cells are required for osteogenic differentiation of these cells during adult bone homeostasis and repair, whereas lack of ANK functions favors adipogenic differentiation.

show abstract

“…Cells differentiation into osteoblasts was investigated by ALP assay that is a marker of early stages of osteogenic differentiation. ALP plays a role in the conversion of inorganic pyrophosphate into inorganic phosphate, a process that promotes the formation of bone matrix, before the mineralization process [44]. Over a period of 14 days, all the studied hydrogels promoted cell differentiation at the same level.…”

Section: Discussionmentioning

confidence: 96%

Sustained Release from Injectable Composite Gels Loaded with Silver Nanowires Designed to Combat Bacterial Resistance in Bone Regeneration Applications

Mori¹,

Hafidh²,

Mele³

et al. 2019

Preprint

View full text Add to dashboard Cite

One-dimensional nanostructures such as silver nanowires (AgNWs) have attracted considerable attention owing to their outstanding electrical, thermal and antimicrobial properties; however, their application in the prevention of infections linked to bone tissue regeneration interventions has not yet been explored. Here we report on the development of an innovative scaffold prepared from chitosan, composite hydroxyapatite and AgNWs (CS-HACS-AgNWs) having both bioactive and antibacterial properties. In vitro results highlighted the antibacterial potential of AgNWs against both gram-positive and gram-negative bacteria. The CS-HACS-AgNWs composite scaffold demonstrated suitable Ca/P deposition, improved gel strength, reduced gelation time, and sustained Ag + release within therapeutic concentrations. Antibacterial studies showed that the composite formulation was capable of inhibiting bacterial growth in suspension and of completely preventing biofilm formation on the scaffold in the presence of resistant strains. The hydrogels were also shown to be biocompatible, allowing cell proliferation. In summary, the developed CS-HACS-AgNWs composite hydrogels demonstrated significant potential as a scaffold material to be employed in bone regenerative medicine, as they present enhanced mechanical strength combined with the ability to allow calcium salts deposition, while efficiently decreasing the risk of infections. The results presented justify further investigations into potential clinical applications of these materials.to the bulk materials due to their reduced dimensions, controlled structure and large surface-tovolume ratio [3].One, two, and three-dimensional silver nanostructures have been previously reported, with many studies focusing on silver nanowires (AgNWs). AgNWs are anisotropically grown to obtain an aspect ratio higher than 10 and are typically 10-200 nm in diameter and 5-100 µm in length [2]. To date, silver nanoparticles (AgNPs) have been much more extensively studied than AgNWs, and their distinctive high thermal and electrical conductivity, surface-enhanced Raman scattering, chemical stability, catalytic activity and non-linear optical behavior have been highlighted; as well as their antibacterial activity that has extended their usage into several biomedical fields [4]. They are promising agents against a wide range of fungi, viruses and bacterial species; even though the mechanism of action is still not entirely understood, it is believed that nanosilver serves as a source for Ag + ions that can rupture microbial cell walls, denature proteins, block cell respiration and eventually induce cell death [5]. AgNPs have been used as antimicrobial agents in wound dressing [6], coating of catheters [7] and of cardiovascular implants [8], bone cements [9] and dental materials [10]. Currently literature on biomedical applications of AgNWs is still limited [2]; emerging applications include textiles [11], wound coatings [12], drug release and tissue regeneration [13].In this work, we explore for the first time the...

show abstract

Pyrophosphate Stimulates Differentiation, Matrix Gene Expression and Alkaline Phosphatase Activity in Osteoblasts

Cited by 48 publications

References 49 publications

Bone morphogenetic protein 9 induces osteogenic differentiation of germ cell 1 spermatogonial cells

Bone morphogenetic protein 9 induces osteogenic differentiation of germ cell 1 spermatogonial cells

The role of the progressive ankylosis protein (ANK) in adipogenic/osteogenic fate decision of precursor cells

Sustained Release from Injectable Composite Gels Loaded with Silver Nanowires Designed to Combat Bacterial Resistance in Bone Regeneration Applications

Contact Info

Product

Resources

About