Wound dressing composed of hyaluronic acid and collagen containing EGF or bFGF: comparative culture study

Abstract

We developed a novel wound dressing composed of a hyaluronic acid (HA) and collagen (Col) spongy sheet containing epidermal growth factor (EGF) or basic fibrolast growth factor (bFGF) by freeze-drying method (EGF-wound dressing or bFGF-wound dressing, respectively). A wound dressing without any growth factor was prepared as a control in a similar manner as above (C-wound dressing). Intermolecular cross-linkage between Col molecules was induced by UV irradiation. The release behavior of free HA from the wound dressing was investigated using a C-wound dressing. The weight of C-wound dressing after 1 day, 3, 5, and 7 days of incubation on top of a Col gel sheet at the air-water interface (wound surface model) was 55, 36, 30, and 19% of the original weight, respectively. Most free HA and a part of Col was released from the cross-linked Col network in the wound dressing during incubation, as the original Col content in the wound dressing was 33%. Next, fibroblast proliferation was assessed in conventional culture medium preconditioned by immersion of a piece of C-, EGF-, or bFGF-wound dressing, i.e. C-conditioned medium, EGF-conditioned medium, or bFGF-conditioned medium. Cell proliferation in C-conditioned medium increased to approximately the same level as that in conventional medium. Cell proliferation in EGF- and bFGF-conditioned medium was 1.9 times and 2.6 times greater than that in conventional medium after 7 days of cultivation, respectively. Finally, cytokine production of fibroblasts was assessed in a wound surface model using a fibroblast-incorporating Col gel sheet (cultured dermal substitute [CDS]). CDS was elevated to the air-medium interface, on which each wound dressing was placed and cultured for 7 days. Fibroblasts in CDS covered with EGF-wound dressing released 3.6 times more vascular endothelial growth factor (VEGF) and 4.6 times more hepatocyte growth factor (HGF) when compared with the C-wound dressing. Fibroblasts in CDS covered with bFGF-wound dressing released 10.2 times more VEGF and 6.3 times more HGF when compared with the C-wound dressing. This finding indicates that bFGF-wound dressing can facilitate more effectively the VEGF and FGF production compared with EGF-wound dressing.

<i>In Vitro</i> Analysis of VEGF and HGF Production by Fibroblast in Cultured Dermal Substitute Combined with EGF-Incorporating Top Dressing

“…EGF dose dependency appeared to more sensitive in Group I than in Group II, as the sell seeding density in Group I was half that in Group II. In the experiment using ELISA, fibroblasts in CDS in Group I released increased amounts of VEGF and HGF when laminated with membranes containing EGF at 0.1 and 0.2 µg/cm 2 , but failed to increase at 0.5 µg/cm 2 . On the other hand, fibroblasts in CDS in Group II released increased amounts of VEGF and HGF when laminated with a membrane containing EGF ranging from 0.1 to 0.5 µg/cm 2 .…”

“…We have developed various wound dressings and cultured skin substitutes. The wound dressing is composed of hyaluronic acid (HA) and collagen (Col) spongy sheet containing epidermal growth factor (EGF) [1] [2]. The cultured dermal substitute (CDS) is composed of HA and Col spongy sheet containing human fibroblasts [3] [4].…”

Development of anti-adhesive spongy sheet composed of hyaluronic acid and collagen containing epidermal growth factor

“…[17] Briefly, in order to create a wound surface model, a sheet of CDS (4 cm × 5 cm) was placed on a stainless mesh in a 100-cm 2 dish containing 60 mL of conventional culture medium. Each spongy sheet (4 cm × 5 cm) was placed on CDS.…”

“…[1][2][3][4][5][6][7][8][9][10][11][12][13] We have developed several types of wound dressing composed of hyaluronic acid (HA) and collagen (Col) spongy sheet containing epidermal growth factor (EGF). [14][15][16][17] Both HA and Col have the potential to facilitate wound healing. In the development of anti-adhesive products, researchers have focused on preventing surgically excised tissue from adhering to surrounding tissues using mechanical barriers such as membranes and spongy sheets.…”

Bioactive bilayered dressing for compromised epidermal tissue regeneration with sequential activity of complementary agents

“…These bioactive molecules should ideally have pro-angiogenic, anti-microbial, and re-epithelialization properties, and modulate the activity of growth factors involved in regeneration process, such as fibroblast growth factor (FGF) or vascular endothelial growth factor (VEGF) [8], and stem cells [9,10].…”