Fig 1.
Effects of cinnamtannin B-1 on migration of mouse mesenchymal stem cells (MSCs).
(A) Increase in population of Lin−/PDGFRα+/Sca-1+ peripheral blood mononuclear cells of mice after systemic administration of cinnamtannin B-1, n = 6 per group. *P < 0.05 vs. control. (B) KUM6 cells localized in wound healing models; ffLuc-expressing KUM6 cells were injected 4 days after surgical incision (day 1). Images of a representative animal are shown on day 2 and 5 after ffLuc-KUM6 cell injection. (C) Quantification of photon counts at wound site on day 2 and 5 (n = 8–9 and 6–7, respectively); **P < 0.01 vs. control.
Fig 2.
Effects of cinnamtannin B-1 in a diabetic mouse wound healing model.
Topical application of cinnamtannin B-1 or phosphate-buffered saline (PBS, control) was repeated three times per week. (A) Time course of quantification of wound size. Percentage (%) of wound area was calculated as wound area at different times/wound area on day 0 × 100; n = 10 in each group, *P < 0.05 and **P < 0.01 vs. control. (B) Representative images of wounds on day 19. (C) Hematoxylin and eosin (H&E) staining of specimens on day 14 (200× magnification). (D) α-SMA immunostaining (200× magnification). The capillaries are indicated by arrows. (E) Masson Trichrome staining (200× magnification). (F) Number of capillaries in scar area was counted; n = 6, *P < 0.05 vs. control. (G) Granulation tissue area (mm2) was calculated as blue stained portion in wound; n = 6, **P < 0.01 vs. control.
Fig 3.
Effects of specific inhibitors on cinnamtannin B-1-induced migration of KUM6 cells.
(A) Phosphatidylinositol 3-kinase (PI3K) inhibitor, LY294002. (B) Phospholipase C (PLC) inhibitor, U-73343. (C) Protein kinase C (PKC) inhibitor, Gö6983. (D) Lipoxygenase (LOX) inhibitor, nordihydroguaiaretic acid. (E) Purine inhibitor, azathioprine. Values are mean ± SE while migration indices are normalized to that of cinnamtannin B-1-treated cells; n = 4/group, **P < 0.01 vs. control.
Fig 4.
Effects of various flavonoids and polyphenols on KUM6 cells.
(A) Structure of cinnamtannin B-1 and MSC migration and proliferation after stimulation with cinnamtannin B-1, (B) procyanidin A2, (C) procyanidin B1, (D) morelloflavone, (E) cupressuflavone, (F) (+)-catechin, (G) genistein, (H) caffeic acid, and (I) gallic acid. Cell migration assay values are means ± SE and indices were normalized to platelet-derived growth factor (PDGF)-BB-treated cells; n = 4/group, *P < 0.05 and **P < 0.01 vs. control. Cell proliferation assay values are mean ± SE percentages normalized to that of untreated controls; n = 6/group, *P < 0.05 and **P < 0.01 vs. control.
Fig 5.
Effects of specific inhibitors on biflavonoid-induced migration of KUM6 cells.
(A) Procyanidin A2, (B) procyanidin B1, and (C) morelloflavone. Inhibitors were used at 25 μM, 50 nM, 1 μM, 25 μM, and 100 μM for LY294002, Gö6983, U-73343, nordihydroguaiaretic acid, and azathioprine, respectively. Values are mean ± SE of migration indices normalized to that of biflavonoid-treated cells; n = 4/group, **P < 0.01 vs. control.