Angiogenic Growth Factor Synergism in a Murine Tissue Engineering Model of Angiogenesis and Adipogenesis

Abstract

De novo tissue generation stimulated by three angiogenic growth factors administered in a factorial design was studied in an in vivo murine tissue engineering chamber. A silicone chamber was implanted around the epigastric pedicle and filled with Matrigel with 100 ng/ml of recombinant mouse vascular endothelial growth factor-120 (VEGF 120 ), recombinant human basic fibroblastic growth factor (FGF-2), or recombinant rat platelet-derived growth factor-BB (PDGF-BB) added as single, double, or triple combinations. Angiogenesis, supporting tissue ingrowth, and adipogenesis were assessed at 2 and 6 weeks by immunohistochemistry and morphometry. At 2 weeks angiogenesis was synergistically enhanced by VEGF 120 ؉ FGF-2 (P ‫؍‬ 0.019). FGF-2 (P ‫؍‬ 0.008) and PDGF-BB (P ‫؍‬ 0.01) significantly increased connective tissue/inflammatory cell infiltrate (macrophages, pericytes, and preadipocytes) in double and triple combinations compared with control. At 6 weeks sequential addition of growth factors increased the percent volume of adipose tissue (P < 0.0005, each main effect), with a synergistic increase in adipose tissue in combination treatments (P < 0.0005). Groups containing 300 ng/ml of single growth factors produced significantly less adipose tissue than the triple growth factor combination (P < 0.0005, VEGF 120 and PDGF-BB; P < 0.001, FGF-2). In conclusion, angiogenic growth factor combinations increased early angiogenesis and cell infiltration resulting in synergistically increased adipose tissue growth at 6 weeks. Two way and higher level synergies are likely to be important in therapeutic applications of angiogenic growth factors.

Adipose Injury–Associated Factors Mitigate Hypoxia in Ischemic Tissues through Activation of Adipose-Derived Stem/Progenitor/Stromal Cells and Induction of Angiogenesis

“…Indeed, in some previous reports, superior results for angiogenesis have been obtained through the application of multiple factors than through the introduction of a single factor, eg, paired combinations of bFGF and PDGF, 26,27 VEGF and bFGF, 28,29 VEGF and PDGF, 30 or PDGF and TGF-␤, 31 or a triple combination of bFGF, PDGF, and VEGF. 32 Our findings with incomplete AIC with either two or three growth factors suggest that PDGF, EGF, and TGF-␤ contributed to hASC migration and that bFGF played a pivotal role in network formation, although these effects were likely augmented by the synergism of all of the factors. Although the reproducible methods for inducing hASC Wholemount histologic analysis of living adipose tissue.…”

The role of adipose protein derived hydrogels in adipogenesis

“…FGF-2 has enjoyed significant popularity as a potential regulator for adipose formation when Matrigel is implanted in small animal models [3,17]. The levels of FGF-1 and FGF-2 in the extracts were quantified using sandwich ELISAs.…”

“…Rophael et al investigated the effects of angiogenic growth factors including vascular endothelial growth factor, FGF-2 and platelet derived growth factor-BB in an in vivo murine model in which implanted chambers were filled with growth factor-reduced MatrigelÔ supplemented with growth factor combinations. The study indicated that MatrigelÔ with angiogenic growth factors promoted neovascularization and adipose tissue formation [17]. Studies have also shown that supplementation of MatrigelÔ with an adipose graft promotes rapid adipogenesis by host cells [15].…”

Application of Platelet-Rich Plasma and Platelet-Rich Fibrin in Fat Grafting: Basic Science and Literature Review

“…Rophael et al, found the cocktail of angiogenic growth factors (VEGF, FGF, PDGF-BB) does not only enhance early angiogenesis, but also late de novo adipogenesis compared to each single growth factor in a murine tissue engineering model. 45 Hence, the angiogenic effect of PRP might not only maintain the survival of mature fat cells, but also induce de novo adipogenesis.…”