The proliferation and differentiation of osteoblasts in co-culture with human umbilical vein endothelial cells: An improved analysis using fluorescence-activated cell sorting

Zhang, Yu; Schedle, Andreas; Matĕjka, M; Rausch‐Fan, Xiaohui; Andrukhov, Oleh

doi:10.2478/s11658-010-0026-0

Cited by 21 publications

(21 citation statements)

References 37 publications

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“…22,32 The evidence that ALP activity is promoted by the presence of EC in coculture is very strong. 2,[7][8][9]13,15,19,29,33 In our study, ALP activity was significantly higher in all investigated OB:EC proportions compared with the HBDC monoculture on day 7 (see Fig. 4).…”

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confidence: 67%

“…9 Despite all these ambiguities, it is interesting that the relation between OB monoculture and OB-EC coculture with respect to all the investigated osteogenic genes expression that we have obtained is similar to the results reported for MG63 osteosarcoma cell line. 15 Clearly, this aspect needs further investigation.…”

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confidence: 99%

“…13 At the same time, recent studies highlighted the stimulating influence of EC on alkaline phosphatase (ALP) activity in OB. 2,6,7,14,15 The effect of EC on the induction of osteoblastic differentiation markers in osteoprogenitor cells, such as runt-related transcription factor 2 (Runx2), ALP, and osteocalcin represents another intensively investigated processes. 6,13,16 Our knowledge about EC influence on OB differentiation remains far from complete.…”

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confidence: 99%

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Contribution of Endothelial Cells to Human Bone-Derived Cells Expansion in Coculture

Leszczyńska

Żyżyńska-Granica

Koziak

et al. 2013

Tissue Engineering Part A

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Creating a functional vascularized bone tissue remains one of the main goals of bone tissue engineering. Recently, a growing interest in the crosstalk between endothelial cells (EC) and osteoblasts (OB), the two main players in a new bone formation, has been observed. However, only a few reports have addressed a mutual influence of OB and EC on cell proliferation. Our study focuses on this issue by investigating cocultures of human bone-derived cells (HBDC) and human umbilical vein endothelial cells (HUVEC). Three various proportions of cells have been used that is, HBDC:HUVEC 1:1, 1:4, and 4:1 and the cocultures were investigated on day 1, 4, and 7, while HUVEC and HBDC monocultures served as reference. We have detected enhanced alkaline phosphatase (ALP) activity in a direct HBDC-HUVEC coculture. This effect was not observed when cells were separated by an insert, which is consistent with other reports on various OB-EC lineages. The appearance of gap-junctions in coculture was confirmed by a positive staining for connexin 43. The number of cells of both phenotypes has been determined by flow cytometry: CD-31-positive cells have been considered EC, while CD-31-negative have been counted as OB. We have observed an over 14-fold increase in OB number after a week in the 1:4 HBDC:HUVEC coculture as compared with less than fourfold in monoculture. The increase in HBDC number in 1:1 coculture has been less pronounced and has reached the value of about sevenfold. These results correspond well with the cell proliferation rate, which has been measured by 5-bromo-2¢-deoxyuridine incorporation. Moreover, at day 7 EC have been still present in the coculture, which is inconsistent with some other reports. Real-time polymerase chain reaction analysis has revealed the upregulation of ALP and collagen type I genes, but not osteocalcin gene, in all the cocultures grown without pro-osteogenic additives. Our study indicates that HUVEC significantly promote HBDC expansion and upregulate collagen I gene expression in these cells. We believe that these findings have application potency in bone tissue engineering.

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confidence: 67%

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confidence: 99%

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confidence: 99%

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Contribution of Endothelial Cells to Human Bone-Derived Cells Expansion in Coculture

Leszczyńska

Żyżyńska-Granica

Koziak

et al. 2013

Tissue Engineering Part A

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“…Furthermore, Zang et al 25 Nevertheless, the genetic profile of co-cultured HMSC and HDMEC, after being sorted, was distinct from that observed in the respective control monoculture, regarding the level of expression and timedependent profile.…”

Section: Discussionmentioning

confidence: 96%

Response of Monocultured and Co-Cultured Human Microvascular Endothelial Cells and Mesenchymal Stem Cells to Macroporous Granules of Nanostructured-Hydroxyapatite Agglomerates

Laranjeira¹,

Fernandes²,

Monteiro³

2013

Journal of Biomedical Nanotechnology

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Recent studies have shown that hydroxyapatite (HA) nanocrystalline have better functional properties that are important to create suitable local conditions for bone formation, when implanted in an osseous environment. Bone formation depends on several complex processes, including a tight communication between endothelial cells and osteoblasts and mesenchymal stem cells. This study examined the interaction between human dermal microvascular endothelial cells (HDMEC) and human mesenchymal stem cells (HMSC), in monoculture and co-culture on,macroporous granules of nanostructured-hydroxyapatite agglomerates. Cell viability/proliferation was assessed through MTT and DNA quantification assays. CLSM and SEM observations allow the study of cell morphology and growth pattern of cells. The angiogenic and osteogenic genes expression were studied using real time PCR and cell differentiation was assessed by ALP activity and matrix mineralization assays. Matrigel tube-like formation assay was also used. Increased expression levels of genes related with osteogenesis and angiogenesis was evident. The osteoblastic phenotype was clearly promoted, as evidenced by the over-expression of osteoblastic genes, increased ALP activity and matrix mineralization. The work clearly demonstrated that the nanostructured-HA granules were able to support cell type's survival, proliferation and individual functionality in a monoculture and co-culture system, for 21 days. HMSC seeded on the granules were able to differentiate into osteoblastic phenotype. The results achieved suggest that nano-structured HA granules may be considered promising implants for bone regeneration and tissue engineering application, in which the granules can be pre-seeded with these two types of autologous cells, before bone graft implant.

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“…Vascularization is recognized to play a central role in normal bone tissue homeostasis and repair [6]. The processes integral to bone formation are thought to be dependent on angiogenesis due to the close interaction between the bone-forming osteoblasts (OBs) and the vessel-forming endothelial cells (ECs) [7][8][9][10].…”

Section: Introductionmentioning

confidence: 99%

Influence of scaffold properties on the inter-relationship between human bone marrow derived stromal cells and endothelial cells in pro-osteogenic conditions

et al. 2015

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The proliferation and differentiation of osteoblasts in co-culture with human umbilical vein endothelial cells: An improved analysis using fluorescence-activated cell sorting

Cited by 21 publications

References 37 publications

Contribution of Endothelial Cells to Human Bone-Derived Cells Expansion in Coculture

Contribution of Endothelial Cells to Human Bone-Derived Cells Expansion in Coculture

Response of Monocultured and Co-Cultured Human Microvascular Endothelial Cells and Mesenchymal Stem Cells to Macroporous Granules of Nanostructured-Hydroxyapatite Agglomerates

Influence of scaffold properties on the inter-relationship between human bone marrow derived stromal cells and endothelial cells in pro-osteogenic conditions

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