Fig 2: Hemocompatibility and cytocompatibility assay of the purified Bm-ponericin-L1 from infected B. mori larvae on erythrocytes (a) and primary fibroblast cell lines (ATCC PCS-201-012) (b) respectively. The data were expressed as mean ± standard deviation and the significance level was set at p < 0.05.

Fig 3: Size of keratinocytes grown on different substrates. (A) (i) Phase contrast images show differences in the size of keratinocytes grown on the various substrates. Images were captured on day 3 of culture. Scale bars are 100 µm. (ii) Representative images of keratinocytes stained with phalloidin-Alexa Fluor 488 demonstrating that phalloidin staining accurately revealed keratinocyte size. Scale bars are 50 µm. Nuclei were stained using DAPI (Blue). (B) Frequency of keratinocytes of differing size. Cells were assigned into categories based on cell area (small < medium < large = <2000 µm2 < 4000 µm2 < 6000 µm2), where M = FibMat, C = Col I and T = TCP. The data are the percent of total cells. Data from three separate experiments using Fib-Mats from ATCC PCS-201-012 are shown. *p = 0.01, but replicate experiments using Fib-Mats from EBL028 gave very similar data.

Fig 4: Colony forming ability of keratinocytes previously cultured on different substrates. (A) Representative image of the colonies formed by keratinocytes previously cultured on different substrates. Keratinocytes were cultured for three days on Fib-Mat from ATCC PCS-201-012, collagen I or uncoated TCP, the cells were harvested and single cells were seeded at low density onto mitomycin C-treated 3T3-J2 feeder cells, with 3 replicate wells for keratinocytes from each substrate. After 12 days keratinocyte colonies were stained with toluidine blue and imaged. Replicate experiments performed with Fib-Mat from EBL028 and collagen I produced very similar data. (B) Quantification of the colonies. All colonies were counted and categorised: large colonies were = 1mm2, and small colonies were cell clusters <1 mm2. The data from 3 replicate wells are shown, and are representative of three separate experiments. *p < 0.01

Fig 5: ECM proteins deposited by dermal fibroblasts (A) The effect of MMC on HDF ECM deposition. The cells and matrix were immunostained for collagen I (i, v), collagen IV (ii, vi), fibronectin (iii, vii) and perlecan (iv, viii). Scale bars are 100 µm. (B) The 3D architecture of the ECM deposited by HDF before and after decellularisation with EDTA, ammonium hydroxide (AH) or phospholipase A2 (PLA2) as revealed by collagen I immunostaining and DAPI staining. Scale bars are 10 µm. (C) The thickness of the ECM after decellularisation. The colour coding represents the Z-depth location within the 3D z-stacked image. Shown are data for ATCC PCS-201-012 ECM, but EBL028 derived ECM was of a similar thickness of 4–5 µm.