Table 1.
Primers used for real-time RT-PCR analysis.
Figure 1.
Structural properties of ultralow temperature broken ANBP powder.
(A) Scanning electron microscope micrographs of ultralow temperature broken and conventional ANBP powders. (B) Static laser scattering analysis of dissolved ANBP particle size distribution(C) Liquid chromatography chromatograms of two different methods of ANBP powder preparation.
Figure 2.
ANBP accelerates wound healing and inhibits scar formation.
(A) Representative photographs of full-thickness skin wounds at various time points after treatment with or without ANBP powders. Scale Bar = 5 mm. (B) Time course of wound closure for each experimental and control group (n = 6 for each group, repeated 3 times in total). (C) Comparing of wound healing time after treatment with or without ANBP powders. **p<0.01.
Figure 3.
Histological characteristics of wound healing and scar formation after ANBP treatment.
(A) Histological observation of wound healing at 4, 7, 14 and 28 days post-wounding with or without ANBP treatment. G: Granulation tissue, ET: Epithelial tongue. Scale bars = 100 µm. (B) Quantitative analysis of granulation tissue thickness after 4 days post-wounding. * p<0.05. (C) Quantitative analysis of crawling distance after 4 days post-wounding. ** p<0.01. (D) The thin∶thick ratio of epithelial thickness at 28 days post-wounding. * p<0.05. (E) The scar elevation index (SEI) was significantly decreased on ANBP side at 28 days post-wounding. ** p<0.01.
Figure 4.
Effect of ANBP on extracellular matrix (ECM) synthesis.
(A) Histopathological observation and masson staining of collagen in wound healing at 7, 14 and 28 days post-wounding with or without ANBP treatment. Scale bars = 100 µm. (B) Sirius red staining of collagen in wound healing at 7, 14 and 28 days post-wounding with or without ANBP treatment. Scale bars = 100 µm.
Figure 5.
Changes in expression of inflammatory factors and collagen after ANBP treatment.
mRNA levels were detected by quantitative RT-PCR at 4, 7, 14 and 28 days post-wounding. mRNA levels are shown for: (A) IL-1; (B) TNF-α; (C) Col I; (D) Col III. Expression levels were calculated using standard curves and normalized to expression level of β-actin as an endogenous reference gene. * p<0.05, ** p<0.01.
Figure 6.
Effect of ANBP on TGF-β1/Smad Pathway.
(A) Immunohistochemistry for TGF-β1 in the wound granulation tissue at 7 and 14 days post-wounding. Scale bar = 50 µm. Expressions of Smad2 (B) and Smad3 (C) were detected by western blot at 4, 7, 14 and 28 days post-wounding. mRNA levels were detected by quantitative RT-PCR at 4, 7, 14 and 28 days post-wounding. mRNA levels are shown for: (D) TGF-β1; (E) Smad2; (F) Smad3. * p<0.05, ** p<0.01.