The Effect of Topical Administration of an Ointment Prepared From Trifolium Repens Hydroethanolic Extract on the Acceleration of Excisional Cutaneous Wound Healing

Seied Kiavash Habibi Zadeh, DVM; Mohammad-Reza Farahpour, DVM, DVSc; Hamed Hamishe Kar, PhD

Wounds. 2020;32(9):253-261.

Conclusions

The results of the current study showed high flavonoid and phenol contents in TRHE reduced the inflammation-induced degenerative effects. Moreover, administration of T repens increased mast-cell distribution and intracytoplasmic carbohydrate ratio, promoted angiogenesis, and shortened the wound healing process. Natural agents are safer than synthetic ones and capable of treating acute wounds without infection. The extract is economical and can be easily prepared. T repens can be recommended as natural compounds for preparing an ointment for wound healing; however, the results are preliminary and based on a rat model with no disease and an acute wound.

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Table 1. Effects of *Trifolium repens* on the area of circular excisional wound contraction
GROUP	WOUND AREA (mm²)±SD
GROUP	d 3	d 6	d 9	d 12	d 15	d 18	d 21
Control	524.7±36.01 (-6.9%)^a	386.8±15.6 (21.1%)^a	241.5±23.6 (50.7%)^a	152.7±24.9 (68.8%)^a	73.5±14.6 (85%)^a	31.0±6.5 (93.6%)^a	13.1±4.5 (97.2%)^a
1.5% T repens	324.9±14.4 (33.7%)^b	209.5±12.3 (57.2%)^b	112.4±33.7 (77%)^b	69.4±20.7 (85.8%)^b	23.9±6.9 (95.1%)^b	11.1±7.6 (97.7%)^b	4.5±1.6 (98.9%)^a
3% T repens	287.5±19.3 (41.4%)^b	149.1±15.0 (69.6%)^c	81.6±20.4 (83.3%)^c	21.1±5.7 (95.6%)^c	16.6±5.9 (96.6%)^b	7.8±3.2 (98.4%)^b	1.7±1.3 (99.7%)^b
6% T repens	283.3±18.3 (42.2%)^b	120.1±24.6 (75.4%)^d	65.1±11.1 (86.7%)^c	26.4±4.3 (94.6%)^c	17.2±8.7 (96.4%)^b	10.2±6.9 (97.9%)^b	1.9±0.4 (99.6%)^b

SD: standard deviation; d: day
All data are expressed as mean±SD.
^{a, b, c}Represents significant differences between marked data in the same day (P<.05).

Table 2. Histopathologic and histomorphometric analysis
DAY	GROUP	EDEMA SCORE	IMMUNE CELLS	FIBROBLAST/FIBROCYTE	COLLAGEN SCORE	EPIDERMIS THICKNESS	PAS
3	Control	++++	80.0±3.6^a	0.5±0.5^a	-	-	-
	1.5% T repens	++	27.0±5.2^b	4.6±1.0^b	-	-	-
	3% T repens	+++	24.0±7.0^b	4.3±1.5^b	-	-	-
	6% T repens	++	27.0±2.0^b	11.0±1.0^c	-	-	+
7	Control	++++	52.0±2.5^a	11.6±1.0^a	+	0^a	+
	1.5% T repens	++	30.0±5.0^b	19.6±1.5^b	++	0^a	+
	3% T repens	+	35.0±4.5^b	27.3±2.5^c	++	0^a	++
	6% T repens	-	22.3±3.2^c	31.6±1.5^d	++	35.4±3.6^b	++
14	Control	+++	35.0±1.5^a	15.0±1.5^a	+	10.3±3.4^a	+
	1.5% T repens	++	11.0±1.0^b	24.6±1.5^b	++	36.3±5.7^b	++
	3% T repens	-	8.0±1.5^c	28.3±1.5^c	+++	64.4±3.6^c	+++
	6% T repens	-	7.0±2.6^c	30.0±1.0^b	++++	83.6±10.3^d	+++
21	Control	-	19±1.7^a	7.0±1.0^a	++	20.4±4.1^a	+
	1.5% T repens	-	7±1.0^b	15.0±3.0^b	+++	73.2±4.3^b	+++
	3% T repens	-	4.6±0.6^c	18.0±0.9^c	+++	90.0±6.8^c	++++
	6% T repens	-	4.3±05^c	22.3±1.5^d	++++	120.2±5.3^d	++++

PAS: periodic acid-Schiff; T repens: Trifolium repens
Note: The Masson trichrome staining was scored as ++++: intensive; +++: moderate; ++: moderate to mild; +: mild; and -: negative.
^{a, b, c, d}Represents significant differences (P<.05) between the marked data in the same column and day.

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The Effect of Topical Administration of an Ointment Prepared From Trifolium Repens Hydroethanolic Extract on the Acceleration of Excisional Cutaneous Wound Healing

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