Comparative study assessing effects of sonic hedgehog and VEGF in a human co-culture model for bone vascularisation strategies

Dohle, Eva; Fuchs, Sabine; Kolbe, M.; Hofmann, Alexander; Schmidt, Harald; Kirkpatrick, Charles James

doi:10.22203/ecm.v021a12

Cited by 40 publications

(40 citation statements)

References 62 publications

(48 reference statements)

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“…This model was used in previous studies to assess prevascularization strategies, the effects of biomaterial modification on bone tissue engineering, and the effect of the morphogen sonic hedgehog (SHH), to promote and accelerate angiogenesis. 21,22 In the present work, we show that MHS-increased angiogenesis in a co-culture system is mediated via an increase in angiogenic factors produced by the osteoblasts. Moreover, we have confirmed that some HSPs and HSP-related angiogenic and osteogenic factors are involved in vascularization and bone regeneration processes.…”

Section: Introductionsupporting

confidence: 54%

Mild Heat Stress Enhances Angiogenesis in a Co-culture System Consisting of Primary Human Osteoblasts and Outgrowth Endothelial Cells

Fuchs²,

Böse³

et al. 2014

Tissue Engineering Part C: Methods

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The repair and regeneration of large bone defects, including the formation of functional vasculature, represents a highly challenging task for tissue engineering and regenerative medicine. Recent studies have shown that vascularization and ossification can be stimulated by mild heat stress (MHS), which would offer the option to enhance the bone regeneration process by relatively simple means. However, the mechanisms of MHS-enhanced angiogenesis and osteogenesis, as well as potential risks for the treated cells are unclear. We have investigated the direct effect of MHS on angiogenesis and osteogenesis in a co-culture system of human outgrowth endothelial cells (OECs) and primary osteoblasts (pOBs), and assessed cytotoxic effects, as well as the levels of various heat shock proteins (HSPs) synthesized under these conditions. Enhanced formation of microvessel-like structures was observed in co-cultures exposed to MHS (41°C, 1 h), twice per week, over a time period of 7-14 days. As shown by real-time polymerase chain reaction (PCR), the expression of vascular endothelial growth factor (VEGF), angiopoietin-1 (Ang-1), angiopoietin-2 (Ang-2), and tumor necrosis factor-alpha was up-regulated in MHS-treated co-cultures 24 h post-treatment. At the protein level, significantly elevated VEGF and Ang-1 concentrations were observed in MHS-treated co-cultures and pOB mono-cultures compared with controls, indicating paracrine effects associated with MHS-induced angiogenesis. MHS-stimulated co-cultures and OEC mono-cultures released higher levels of Ang-2 than untreated cultures. On the other hand MHS treatment of cocultures did not result in a clear effect regarding osteogenesis. Nevertheless, real-time PCR demonstrated that MHS increased the expression of mitogen-activated protein kinase, interleukin-6, and bone morphogenetic protein 2, known as HSP-related molecules in angiogenic and osteogenic regulation pathways. In agreement with these observations, the expression of some selected HSPs also increased at both the mRNA and protein levels in MHS-treated co-cultures.

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Section: Introductionsupporting

confidence: 54%

Mild Heat Stress Enhances Angiogenesis in a Co-culture System Consisting of Primary Human Osteoblasts and Outgrowth Endothelial Cells

Fuchs²,

Böse³

et al. 2014

Tissue Engineering Part C: Methods

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“…12,13 Interestingly, in addition to increasing capillary density, Shh gene therapy in the ischemic myocardium was shown to promote the formation of larger and more muscularized blood vessels. 14 Moreover, Shh was shown to upregulate MC-related markers during the formation of microvessel-like structures in vitro 15 suggesting a potential action of Shh in vessel maturation. At the cellular level, recombinant Shh was shown to induce SMC and pericyte proliferation and survival [16][17][18][19] and to contribute to PDGF-BB-induced myofibroblast proliferation.…”

Section: Introductionmentioning

confidence: 99%

Sonic hedgehog mediates a novel pathway of PDGF-BB–dependent vessel maturation

Yao

Renault

Chapouly

et al. 2014

Blood

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“…Several approaches have been proposed to improve the neovascularisation process of engineered tissues during the past decade, including the use of endothelial cells or endothelial progenitor cells (Grellier et al, 2009;Rivron et al, 2008;Rouwkema et al, 2006), strategies based on the natural interaction of the different cell types during the process of bone regeneration using co-culture systems (Fuchs et al, 2007;Kaigler et al, 2003) or additional treatment or coupling of tissue engineered constructs with proangiogenic growth factors or morphogens (Dohle et al, 2011;Dohle et al, 2010;Zisch et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

“…For a better clinical application it would be useful to improve the beneficial effect of the co-culture on the angiogenic activation of OECs and to accelerate the formation of vessel-like structures observed in the co-culture. Therefore, additional treatment of co-cultures with vascular endothelial growth factor (VEGF) or the morphogen sonic hedgehog (Shh) result in pro-angiogenic activation as early as after 24 h (Dohle et al, 2011;Dohle et al, 2010). Nevertheless, treating co-cultures with growth factors or morphogens has several limitations and drawbacks, e.g.…”

Section: Introductionmentioning

confidence: 99%

Macrophage-mediated angiogenic activation of outgrowth endothelial cells in co-culture with primary osteoblasts

Dohle

Bischoff²,

Böse³

et al. 2014

eCM

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The successful vascularisation of complex tissue engineered constructs for bone regeneration is still a major challenge in the field of tissue engineering. In this context, co-culture systems of endothelial cells and osteoblasts represent a promising approach to advance the formation of a stable vasculature as well as an excellent in vitro model to identify factors that positively influence bone healing processes, including angiogenesis. Under physiological conditions, the activation phase of angiogenesis is mainly induced by hypoxia or inflammation. Inflammatory cells such as macrophages secrete proinflammatory cytokines and proangiogenic growth factors, finally leading to the formation of new blood vessels. The aim of this study was to investigate if macrophages might positively influence the formation of microvessel-like structures via inflammatory mechanisms in a co-culture system consisting of human outgrowth endothelial cells (OECs) and primary osteoblasts. Treatment of co-cultures with macrophages (induced from THP-1) resulted in a higher number of microvessel-like structures formed by OECs compared to the co-culture. This change correlated with a significantly higher concentration of the proangiogenic VEGF in cell culture supernatants of triple-cultures and was accompanied by an increase in the expression of different proinflammatory cytokines, such as IL-6, IL-8 and TNFα. In addition, the expression of E-selectin and ICAM-1, adhesion molecules which are strongly involved in the interaction between leukocytes and endothelial cells during the process of inflammation was also found to be higher in triple-cultures compared to the double co-cultures, documenting an ongoing proinflammatory stimulus. These results raise the possibility of actively using pro-inflammatory stimuli in a tissue engineering context to accelerate healing mechanisms.

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Comparative study assessing effects of sonic hedgehog and VEGF in a human co-culture model for bone vascularisation strategies

Abstract: The morphogen sonic hedgehog (Shh) seems to mediate adult repair processes in bone regeneration and vascularisation.

Cited by 40 publications

References 62 publications

Mild Heat Stress Enhances Angiogenesis in a Co-culture System Consisting of Primary Human Osteoblasts and Outgrowth Endothelial Cells

Mild Heat Stress Enhances Angiogenesis in a Co-culture System Consisting of Primary Human Osteoblasts and Outgrowth Endothelial Cells

Sonic hedgehog mediates a novel pathway of PDGF-BB–dependent vessel maturation

Macrophage-mediated angiogenic activation of outgrowth endothelial cells in co-culture with primary osteoblasts

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