Accelerative effect of topical Zataria multiflora essential oil against infected wound model by modulating inflammation, angiogenesis, and collagen biosynthesis

Figures & data

Figure 1. GC-MS chromatogram of the essential oil of Zataria multiflora.

Table 1. Chemical constituents of Zataria multiflora essential oil.

No.	Compound	RI-Cal^a	RI-Lit^b	%
1	α-Thujene	926	924	0.4
2	α-Pinene	933	932	2.6
3	Camphene	949	946	0.1
4	β-Pinene	977	974	0.3
5	3-Octanone	982	979	1.2
6	Myrcene	988	988	1.1
7	3-Octanol	992	988	0.2
8	α-Phellandrene	1005	1002	0.2
9	α-Terpinene	1016	1014	2.5
10	p-Cymene	1025*	1020	9.1*
11	Limonene	1028	1024	0.7
12	1,8-Cineole	1031	1026	0.6
13	γ-Terpinene	1059*	1054	8.1*
14	cis-Sabinene hydrate	1067	1065	0.5
15	Terpinolene	1089	1086	0.1
16	Linalool	1099	1095	1.4
17	Borneol	1170	1165	0.2
18	Terpinen-4-ol	1181	1174	0.6
19	α-Terpineol	1198	1186	0.5
20	Thymol methyl ether	1237	1232	2.1
21	Carvacrol methyl ether	1246	1241	0.4
22	Thymol	1303*	1289	52.9*
23	Carvacrol	1308*	1298	6.8*
24	Thymol acetate	1355	1349	1.0
25	(E)-Caryophyllene	1424	1417	3.8
26	Aromadendrene	1442	1439	0.4
27	α-Humulene	1456	1452	0.2
28	Viridiflorene	1496	1496	0.3
29	Spathulenol	1581	1577	0.6
	Monoterpene hydrocarbons			26.2
	Oxygenated monoterpenes			67.4
	Sesquiterpene Hydrocarbons			4.6
	Oxygenated sesquiterpenes			1.4
	Total			99.6

^a RI-Cal: retention indices calculated based on C₆–C₂₄ n-alkenes from a DB-5 column.

^b RI-Cal: retention indices retrieved from literature (Adams 2007).

* Bold values show major compounds in the essential oil.

Figure 2. The effects of topical administration of ZMEO on (B) wound contraction rate and (C) tissue bacterial count (CFU/g) in different days. Superscripts (a–d) indicate significant differences in same day at p < 0.05.

Figure 3. Immunohistochemical staining for angiogenesis:; (A) NCG group, (B) MG group, (C and D) 2 and 4% ZMEO-treated groups. See elevated angiogenesis in ZMEO-treated group (arrows) on day 7 after wound induction. CD 31 staining, 400×.

Figure 4. Cross section from wound area: (A) NCG group, (B) MG group, (C and D) 2 and 4% ZMEO-treated groups. Note well-formed collagen deposition in cross sections from ZMEO-treated animals on day 7 after wound induction (first row), which is significantly increased on day 14 after injury (second row). The third row represents the software analyze for collagen intensity. The cross sections from animals in ZMEO-treated groups exhibited condense collagen deposition versus NCG and MG groups. Masson-trichrome staining and fluorescent staining for collagen, 400× magnification.

Figure 5. Cross section from wound area: (A) NCG group, (B) MG group, (C and D) 2 and 4% ZMEO-treated groups. See well re-epithelialization in ZMEO-treated animals. The re-epithelialization initiated on day 7 after wound induction in ZMEO-treated animals. However, the cross sections from NCG and MG groups are not representing epithelialization. Note well-organized dermis and complete epithelialization with well-formed papillae in ZMEO-treated animals in comparison to NCG and MG groups. Masson-trichrome staining, 100×.

Table 2. The effects of ZMEO on histopathological parameters in different days.

	Edoema	Immune cells	New vessels (mm^2)	Fibroblast (mm^2)	Collagen	Epithelium (µm)
			Day 3
NCG	++++	125.41 ± 4.51^a	0.25 ± 0.13^d	0.73 ± 0.23^d	–	–
MG	+++	109.11 ± 7.24^b	1.14 ± 0.21^c	1.58 ± 0.41^c	–	8.71 ± 1.11^c
2% ZMEO	+++	82.21 ± 7.31^c	1.81 ± 0.11^b	2.35 ± 0.57^b	+	13.21 ± 1.21^b
4% ZMEO	++	63.12 ± 7.51^d	2.30 ± 0.21^a	3.50 ± 0.21^a	++	25.41 ± 1.71^a
			Day 7
NCG	+++	93.12 ± 5.41^a	3.41 ± 0.51^d	1.47 ± 0.33^d	+	14.21 ± 3.41^d
MG	++	72.21 ± 4.14^b	8.21 ± 0.74^c	3.92 ± 0.32^c	++	31.24 ± 4.12^c
2% ZMEO	++	65.10 ± 3.13^c	11.24 ± 0.91^b	4.45 ± 0.32^b	++	65.12 ± 3.74^b
4% ZMEO	++	55.10 ± 4.11^d	14.51 ± 0.51^a	7.81 ± 0.42^a	+++	89.12 ± 3.41^a
			Day 14
NCG	+	76.41 ± 3.24^a	3.31 ± 0.51^b	2.23 ± 0.73^d	++	18.51 ± 2.12^d
MG	–	63.11 ± 3.21^b	5.21 ± 1.31^a	4.75 ± 0.23^c	+++	48.51 ± 3.51^c
2% ZMEO	–	41.24 ± 4.21^c	5.81 ± 1.31^a	7.51 ± 0.91^b	+++	99.41 ± 2.13^b
4% ZMEO	–	35.80 ± 7.10^d	5.41 ± 1.51^a	11.32 ± 0.91^a	++++	131.51 ± 8.41^a

Superscripts (a–d) indicate significant differences in same day at p < 0.05.

Figure 6. The effects of topical application of ZMEO on gene expression. Superscripts (a–d) indicate significant differences in same day at p < 0.05.

Figure 7. The effects of topical application of ZMEO on antioxidant properties. Superscripts (a–d) indicate significant differences in same day at p < 0.05.

Adams RP. 2007. Identification of essential oil components by gas chromatography/mass spectrometry. 4th ed. Carol Stream (IL): Allured Publishing Corporation.

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