Angiogenesis and Fibroblast Growth Factors (FGFs) in a Three-Dimensional Collagen Gel Culture

Nagatoro, Takashi; Fujita, Keiko; Murata, Eri; Akita, Masumi

doi:10.2535/ofaj.80.7

Cited by 7 publications

(7 citation statements)

References 35 publications

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“…The present study revealed the up-regulation of FGF-2 following the up-regulation of Egr-1 after both laser and razor injury. In our previous experiment we confirmed the expressions of FGF-9 (Akita et al 2000;Nagatoro et al 2003) and FGF-7 ) during angiogenesis in collagen gels. Pilcher et al (1997) and Miyagi et al (1998) reported that FGF-9 relates to the expression of matrix metalloproteinases (MMPs) and their inhibitors.…”

Section: Discussionsupporting

confidence: 86%

An in vitro model for studying vascular injury after laser microdissection

Fujita

Komatsu

Tanaka

et al. 2005

Histochem Cell Biol

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We have developed an in vitro model for studying vascular injury. After 7-10 days in a three-dimensional collagen gel culture, capillary-like tubes were formed in the collagen gels. We injured these capillary-like tubes with a laser microdissection system or a scrape method with razors and then examined the collagen gel culture by phase contrast and electron microscopy. After laser injury, profuse necrotic cells were observed around the injured capillary-like tubes and within the tubular lumen compared to the razor injury. We then isolated total RNA from these cultures and prepared cDNA for investigations by quantitative real-time reverse transcription polymerase chain reaction (RT-PCR). Quantitative real time RT-PCR revealed the up-regulation of transcription factor early growth response-1 (Egr-1) after both laser and razor injury, accompanied by the up-regulation of fibroblast growth factor-2 (FGF-2), a proangiogenic factor downstream of Egr-1. The effective laser energy is concentrated on the minute focal spot only. These methods provide a useful in vitro model for studying vascular injury.

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Section: Discussionsupporting

confidence: 86%

An in vitro model for studying vascular injury after laser microdissection

Fujita

Komatsu

Tanaka

et al. 2005

Histochem Cell Biol

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“…Conversely, exogenous FGF-9 did increase Vegf expression (11). Furthermore, expression of Fgf-9 has been identified in an angiogenic collagen gel culture model, coinciding with the formation of capillary-like tubes (27). The data obtained from our injury model indicated that Fgf-9 regulates vascularization in long bone repair by triggering Vegf expression.…”

Section: Discussionmentioning

confidence: 58%

Fgf-9 is required for angiogenesis and osteogenesis in long bone repair

Behr

Leucht

Longaker³

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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128

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Bone healing requires a complex interaction of growth factors that establishes an environment for efficient bone regeneration. Among these, FGFs have been considered important for intrinsic bonehealing capacity. In this study, we analyzed the role of Fgf-9 in long bone repair. One-millimeter unicortical defects were created in tibias of Fgf-9 +/− and wild-type mice. Histomorphometry revealed that half-dose gene of Fgf-9 markedly reduced bone regeneration as compared with wild-type. Both immunohistochemistry and RT-PCR analysis revealed markedly decreased levels of proliferating cell nuclear antigen (PCNA), Runt-related transcription factor 2 (Runx2), osteocalcin, Vega-a, and platelet endothelial cell adhesion molecule 1 (PECAM-1) in Fgf-9 +/− defects. μCT angiography indicated dramatic impairment of neovascularization in Fgf-9 +/− mice as compared with controls. Treatment with FGF-9 protein promoted angiogenesis and successfully rescued the healing capacity of Fgf-9 +/− mice. Importantly, although other pro-osteogenic factors [Fgf-2, Fgf-18, and bone morphogenic protein 2 (Bmp-2)] still were present in Fgf-9 +/− mice, they could not compensate for the haploinsufficiency of the Fgf-9 gene. Therefore, endogenous Fgf-9 seems to play an important role in long bone repair. Taken together our data suggest a unique role for Fgf-9 in bone healing, presumably by initiating angiogenesis through Vegf-a. Moreover, this study further supports the embryonic phenotype previously observed in the developing limb, thus promoting the concept that healing processes in adult organisms may recapitulate embryonic skeletal development.tibia | regeneration | tissue B one healing is an efficient regenerative process resulting in newly formed bone equivalent to the original tissue. It involves the interplay of numerous factors over the well-characterized cascade of phases, including inflammation, callus formation, and remodeling (1), which recapitulate aspects of skeletal development (2, 3). Among these factors, several members of the FGF ligands, including FGF-2, -9, and -18, as well their corresponding receptors FGFR1-3, have been identified previously as having a major role during skeletal development (4-12). Although their contribution during skeletal development is well established, less is known about FGFs in fracture healing. The recent identification of different expression patterns of FGF ligands and receptors during fracture repair (13) indicates potential functions in bone regeneration. Within the group of pro-osteogenic FGF ligands, Fgf-9 has been reported to play a role during the development of skeletal vascularization (11).Skeletal vascularization of the injury site is a key step in facilitating successful bone regeneration. Angiogenesis is achieved by two hormonal pathways: the angiopoietin-and VEGF-mediated pathways (2, 14). Vegf was identified previously as a coordinator of chondrocyte growth and bone formation in growth plates of juvenile mice (15). Moreover, Vegf is released physiologically in fracture hematoma (16). C...

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“…Collagen gel culture has been used widely and effectively for analyzing the biological process of angiogenesis [10–13]. Using a three-dimensional (3D) collagen gel culture, we have conducted electron microscopic studies [14, 15] and immunohistochemical studies of fibroblast growth factor- (FGF-) 2 and FGF-9 [16]. Additionally, we have used 3D collagen gel cultures to test angiogenic and anti-angiogenic agents (TNF- α and thalidomide) [17, 18] to study vascular injury after laser microdissection [19] and for profiling of DNA microarray gene expression during angiogenesis [20].…”

Section: Introductionmentioning

confidence: 99%

Detection of the Hematopoietic Stem and Progenitor Cell Marker CD133 during Angiogenesis in Three-Dimensional Collagen Gel Culture

Akita

Tanaka

Matsumoto

et al. 2013

Stem Cells International

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We detected the hematopoietic stem and progenitor cell marker CD133 using immunogold labeling during angiogenesis in a three-dimensional collagen gel culture. CD133-positive cells were present in capillary tubes newly formed from aortic explants in vitro. The CD133-positive cell population had the capacity to form capillary tubes. Lovastatin strongly inhibited cell migration from aortic explants and caused the degradation of the capillary tubes. The present study provides insight into the function of CD133 during angiogenesis as well as an explanation for the antiangiogenic effect of statins.

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Angiogenesis and Fibroblast Growth Factors (FGFs) in a Three-Dimensional Collagen Gel Culture

Cited by 7 publications

References 35 publications

An in vitro model for studying vascular injury after laser microdissection

An in vitro model for studying vascular injury after laser microdissection

Fgf-9 is required for angiogenesis and osteogenesis in long bone repair

Detection of the Hematopoietic Stem and Progenitor Cell Marker CD133 during Angiogenesis in Three-Dimensional Collagen Gel Culture

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