The curative effects of the traditional Chinese herbal medicine "Jinchuang ointment" on excisional wounds

Abstract

Background: "Jinchuang ointment" is a traditional Chinese herbal medicine for external incised wounds. This herbal medicine has been successfully used to treat patients with diabetic foot ulcers and pressure sores in Taiwan for several decades. We previously examined its biological activities on cell-based in vitro assay platforms. Because some patients refused to use animal-derived ingredients ointment during our clinical practice, the efficacy of plant oil-based reconstituted "Jinchuang ointment" was also investigated.

Methods: A porcine excisional wound model was established and used to evaluate its efficacy in vivo in this study. Besides, an unusual clinical case is also present.

Results: As judged from the wound appearance of animal studies on day 14 and the results of blood flow flux at the wound sites on day 28, "Jinchuang ointment" accelerated wound closure significantly better than the control group.

Conclusions: The results from clinical treatment, histopathological evaluation, and the animal study showed that "Jinchung ointment" promotes wound healing significantly better than the control group. Also, sesame oil-reconstituted ointment can be a choice for patients who refuse to use lard-containing ointment.

Keywords: Herbal medicine; Jinchuang ointment; Porcine model; Wound healing.

Fig. 1

a Surgical wound incision, and b wounds on the dorsal area of swine

Fig. 2

Photographs of scalpel wounds on the subject’s hands sutured and treated with neomycin and Jinchuang ointment. Date of photographs: a Sept 5, 2015, 13:47; b Sept 5, 2015, 14:19; c Sept 5, 2015, 14:30; d Sept 7, 2015; e Oct 18, 2015

Fig. 3

Healing process of excisional wounds at day 0, 7, 14, 28 after surgery. a Representative photomicrographs showing the time course of wound healing. b Quantitative data concerning the proportion of the wound remaining open relative to the initial wound area at different time points after surgery. The percentage of unhealed wound area was calculated, taking the value at day 0 as 100%. Data indicate mean ± SD. Through one tailed test analysis, ** represent statistical significance (p value < 0.005) versus the CA group (n = 3)

Fig. 4

Blood flow in the wounded area of porcine at wound sites at day 0, 7, 14, 28 after surgery. a Laser Doppler images showing the time course of wound healing. b The unit of blood flow flux is perfusion unit (PU). We took the value of wound blood flow at day 0 as 100% after surgery. Data indicate mean ± SD. Through one tailed test analysis, * represent statistical significance (p value < 0.05) versus the CA group at day 28 (n = 3)

Fig. 5

Masson Trichrome staining of tissues with different treatments on days 7, 14 and 28, 20×. a CA on day 7, the epithelium did not migrate remarkably from the edge (blackbroken line circle); b TA1 on day 7, the epithelial hyperplasia has migrated less than 50% of the distance (blackbroken line circle); c TA2 on day 7, less than 50% of the epithelium has migrated above the granulation tissue (blackbroken line circle); d TA3 on day 7, less than 50% of the epithelium has migrated above the granulation tissue (blackbroken line circle); e CA on day 14, the epithelial hyperplasia has migrated above the granulation tissue. Only some collagen observed in the granulation tissue; f TA1 on day 14, the fibroblasts have secreted very limited amounts of collagen in the granulation tissue; g TA2 on day 14, similar progress as TA1 on day 14; h TA3 on day 14, similar progress as TA1 on day 14; i CA on day 28, the epithelium was bridged (black circle), only limited collagen labeled in the granulation tissue; j TA1 on day 28, the collagen has crossed the border and has extended to the center (white asterisk), note the epithelium has finished regenerating with epidermal ridge reconstruction; k TA2 on day 28, similar epithelial restoration as TA1 with less collagen in the granulation tissue (white asterisk); l TA3 on day 28, sporadic bridging has occurred (black circle) with less collagen deposition (white asterisk) than TA1

Fig. 6

H&E staining of tissues with different treatments on day 7, 14 and 28, 200×. a CA on day 7, PMNL infiltration with active fibroblast proliferation in the granulation tissue; b TA1 on day 7, similar findings as CA with brownish foreign body debris (black arrow); c TA2 on day 7, similar findings as CA with brownish foreign body debris (black arrow); d TA3 on day 7, similar findings as CA with brownish foreign body debris (black arrow); e CA on day 14, mineralized foreign body debris with diffuse fibroblast infiltration; f TA1 on day 14, diffuse fibroblast infiltration with PMNL withdrawing. Note the brownish foreign body debris still located in the granulation tissue (black arrow); g TA2 on day 14, similar findings as TA1 on day 14 but with more PMNL infiltration; h TA3 on day 14, more PMNL infiltration but less fibroblast infiltration when compared to the TA1 group; i CA on day 28, the wound was refilled without notable PMNL infiltration; j TA1 on day 28, more collagen deposition than CA group. Only a few foreign body debris observed in the granulation tissue (black arrow); k TA2 on day 28, neovascularization present in the granulation tissue with sporadic foreign body debris accumulated (black arrow); l TA3 on day 28, similar findings as TA2 on day 28 with more PMNL infiltration

Fig. 7

Anti-VEGF labelling of tissues with different treatments on day 7, 14 and 28, 200×. a CA on day 7, moderate VEGF secreted from active fibroblasts; b TA1 on day 7, strong and dense VEGF secreted from active fibroblasts; c TA2 on day 7, strong and dense VEGF secreted from active fibroblasts; d TA3 on day 7, moderate VEGF secreted from active fibroblasts; e CA on day 14, negative to weak VEGF secreted from active fibroblasts; f TA1 on day 14, weak VEGF secreted from active fibroblasts; g TA2 on day 14, moderate VEGF secreted from active fibroblasts; h TA3 on day 14, weak to moderate VEGF secreted from active fibroblasts; i CA on day 28, weak VEGF secreted from active fibroblasts; j TA1 on day 28, moderate VEGF secreted from active fibroblasts; k TA2 on day 28, moderate VEGF secreted from active fibroblasts; l TA3 on day 28, weak VEGF secreted from active fibroblasts

Fig. 8

Measurement of cytotoxicity on C2C12 cells treated with “Jinchuang ointment”. a Lard-containing Jinchuang ointment; b Sesame oil-based Jinchuang ointment. Cell numbers were determined with that of the DMSO-treated groups taken as 100%. Through one tailed test analysis, * and ** denote statistical significance (p < 0.05 and p < 0.005, respectively) compared with DMSO and represents two reproducible results