Ointment of Brassica oleracea var. capitata Matures the Extracellular Matrix in Skin Wounds of Wistar Rats

Abstract

Wound healing is a complex process that aims to restore damaged tissue. Phytotherapeutics, such as cabbage, Brassica oleracea var. capitata (Brassicaceae), and sunflower, Helianthus annuus L. (Asteraceae) oil, are used as wound healers. Five circular wounds, each 12 mm in diameter, were made in the dorsolateral region of each rat. The animals were divided into four groups: balsam (B. oleracea); ointment (B. oleracea); sunflower oil (Helianthus annuus); control (saline solution 0.9%). These products were applied daily for 20 days and every four days the tissues of different wounds were removed. The wound contraction area, total collagen, types I and III collagen, glycosaminoglycans, and tissue cellularity were analyzed. In the groups that received ointment and balsam there was reduction in the wound area on days 4, 8, 12, and 20. Throughout the trial period, the balsam and ointment groups showed a higher amount of total collagen, type I collagen, and glycosaminoglycan compared to the others groups. The rats in the groups treated with B. oleracea var. capitata showed a higher number of cells on days 8, 16, and 20. B. oleracea was effective in stimulating the maturation of collagen and increasing the cellularity, as also in improving the mechanical resistance of the newly formed tissue.

Figure 1

Photomicrographs of the scar tissue on days 0, 4, 8, 16, and 20 of the experiment (Sirius Red staining; bar = 60 μm). Tissues were collected every 4 days for 20 days of treatment. Balsam: Brassica oleracea oil; ointment: Brassica oleracea lanolin; sunflower: Helianthus annuus oil; control: 0.9% saline solution.

Figure 2

Cellularity in scar tissue of rats treated with Brassica oleracea and Helianthus annuus. The top panels (a and b) are representative photomicrographs of the computational method of color segmentation to estimate cell distribution in scar tissue (c). Digital images of skin sections stained with hematoxylin and eosin (a, bar = 40 μm) were computationally modified using a color-cube-based mode and a new mask was applied by selecting all pixels of varying dark blue color levels. Then, a new image in black and white was generated after color segmentation (b, bar = 40 μm) and the number of cell nuclei (white color) was measured by point counting. Balsam: Brassica oleracea oil; ointment: Brassica oleracea lanolin; sunflower: Helianthus annuus oil; control: 0.9% saline solution. Data are represented as mean and standard deviation (mean ± SD). ∗ denotes statistical differences between groups (p < 0.05), ∗ versus control; Kruskal-Wallis test.

Figure 3

Representative photomicrographs of tissue cellularity in skin sections of rats observed under light microscope (H&E staining, bars = 25 μm). Tissue fragments were collected from different wounds on days 0, 4, 8, 16, and 20 days. Balsam: Brassica oleracea oil; ointment: Brassica oleracea lanolin; sunflower: Helianthus annuus oil; control: 0.9% saline solution.