Development of Linaria scariosa Herbal Cream: Formulation, In Vivo Wound Healing, and In Vitro Biological Evaluation

Ingredient	Quantity for 100 g	Function
Linaria scariosa extract	200 mg	Active ingredient
Cera alba (White beeswax)	18 g	Thickening agent and consistency enhancer
Paraffin oil	50 g	Oil phase, emollient
Eumulgin B2	0.3 g	Non-ionic emulsifying agent
Cutina GMS	0.8 g	Co-emulsifier and consistency enhancer
Borax (Sodium tetraborate decahydrate)	0.05 g	Emulsion stabilizer and pH-adjusting agent
Distilled water	q.s. to 100g	Aqueous phase

Table 2.

Total phenolic, flavonoid, terpenoid, carotene, and lycopene contents of L. scariosa extract

Classes of bioactive compounds	LS extract
Total phenolic content (μg EAG/mg)	108.98 ± 0.67
Total flavonoid content (μg EQ/mg)	78.77 ± 0 .18
Total terpenoid content (μg EUA/mg)	0.14 ± 0.15
Total carotene content (mg/100 mL)	0.57 ± 0.12
Total lycopene content (mg/100 mL)	0.36 ± 0.08

[i] μg EAG/mg: μg equivalents of gallic acid per mg of dry extract; μg EQ/mg :μg equivalents of quercetin per mg of dry extract; μg EUA/mg: μg equivalents of ursolic acid per mg of dry extract; mg/100 mL: mg of pigment per 100 ml of extract.

Table 3.

Physicochemical, rheological, organoleptic and microbiological properties of the herbal cream

	Parameters	Physical observations after 6 months
Organoleptic properties	Color	Dark yellowish-green color (L= 31.9, a=−11.0 and b*=+19.1)
	Odor	Strong herbal odor
	Homogeneity	Homogenous
	Texture	Viscous, non-greasy and smooth
	Stability	Stable
Physicochemical properties	pH	6.8 ± 0.09 ^a
	Washability	Warm water
	Spreading time	14.91 ± 1.72 ^a S
	Spreadability coefficient (S)	50.07± 5.94 g·cm/s ^a
	Density	1.001±0.02 g/ml ^a
	Moisture content	18.24% ± 0.10 ^a
	Dry matter content	81.75% ± 0.12 ^a
	Ash content	0.02% ± 0.087 ^a
Microbiological stability	Total mesophilic aerobic microorganisms	≤100
	Coagulase-positive Staphylococcus aureus	Complete absence in 1 g
	Escherichia coli	Complete absence in 1 g
	Pseudomonas aeruginosa	Complete absence in 1 g
	Candida albicans	≤100
Rheological properties	Zero-shear viscosity (η₀)	1332 Pa·s
	Flow behavior index (n)	0.7825
	Coefficient of determination (R²)	0.998

a Values expressed are means ± SD of three measurements.

Percentage wound closure for different treated groups over time. C: Control; MDT: Medicament; VE: Vehicle; T: Single dose of the herbal cream; NT: Double dose of the herbal cream.

Table 4.

Antioxidant potential of LS-herbal cream

Sample	Iron chelating power EC₅₀ (μg/mL)^a	DPPH radical scavenging assay IC₅₀ (μg/mL)^a	Permanganate-reducing activity IC₅₀ (μg/mL)^a	Ferric-reducing antioxidant power μg eq AA/mg	Total antioxidant capacity μg eq AA/mg
LS-cream	3.41 ± 0.27	87.63± 0.78	10.38 ± 0.4	0.528 ± 0.02	0.144± 0.01
Ascorbic acid ^b	NT	3.8 ± 0.08	0.42 ± 0.22	NT	NT
BHA	NT	6.73 ± 0.5	NT	0.62 ± 0.09	0.16 ± 0.07
EDTA^b	0.63 ± 0.07	NT	NT	NT	NT

a Values expressed are means ± SD of three measurements (p < 0.05).

b Reference molecule; NT: not tested. μg eq AA/mg: μg equivalents of ascorbic acid per mg of dry extract

Table 5.

Anti-inflammatory activity of the LS cream

	% of inhibition of RBC hemolysis^a
Concentration (mg/mL)	LS-cream	Diclofenac^b
0.25	44.03 ± 0.98	17. 44 ± 1.29
0.5	52.69 ± 0.24	24.98 ± 0.72
1	61.62 ± 0.6	37.77 ± 0.69
2	68.53 ± 1.42	58.96 ± 0.95
4	87.52 ± 0.85	73.51±1.09

a Values expressed are means ± SD of three measurements.

b Reference drug

Table 6.

The in vitro antimicrobial activity of L. scariosa herbal cream

Inhibition zones (mm) and minimum inhibitory concentrations
		Gram-negative strains
Samples	Concentration (μg of extract/g of herbal cream)	E. coli ATCC 25922	Pseudomonas aeruginosa ATCC27853	Klebsiella pneumoniae ATCC70063	M. morganii ATCC25830
LS-cream	2000	15.17 ± 1.18	10.5 ± 2.121	11 ± 1.41	-
	1000	13.42 ± 1.53	9 ±1.414	10 ± 0.94	-
	500	12.10 ± 2.26	8 ± 0.001	6.5 ± 0.71	-
	250	11.05 ± 2.05	6.5 ± 0.707	6 ± 0.71	-
	125	9.00 ± 1.41	-	-	-
	62.5	-	-	-	-
MIC (μg/mL)		125	250	250	-
Penicillin (10 μg/mL)		22.50 ± 0.70	25.00 ± 0.00	23.50 ± 0.70	-
Gentamicin (10 μg/mL)		38.00 ± 0.5	39.00 ± 0.1	27.00 ± 0.5	-
		Gram-positive and fungal strains
Samples	Dilutions	Staphylococcus aureus ATCC 25923	Streptococcus D	Enterococcus spp	Candida albicans ATCC 90029
LS-cream	1/1	15 ± 1.18	13.5 ± 0.12	8.5± 0.23	11 ± 0.03
	1/2	14 ± 1.18	12.5 ± 2.12	7.5 ± 0.71	10.5 ± 0.71
	1/4	11 ± 0.71	12.5 ± 0.54	7.3 ± 0.65	10 ± 0.46
	1/8	10 ± 2.59	12 ± 0.71	7 ± 0.17	9.5 ± 1.41
	1/16	11 ± 0.01	-	-	-
	1/32	8 ± 1.65	-	-	-
MIC (μg/mL)		62.5	250	250	250
Penicillin (10 μg/mL)		26.00 ±1.4	21.00 ±1.41	16 ± 0.7	22 ± 0.8
Gentamicin (10 μg/mL)		34 ± 0.2	27.4 ± 0.32	36.00 ± 1.4	39 ± 0.2

[i] Values expressed are means ± SD of three measurements (P <0.05). (-) No zones of inhibition around the discs. MIC: the minimum inhibitory concentration.

Histological sections with H&E staining showing better healing in T and NT groups compared to other groups of the study, X400. C: Control; VE: Vehicle; MDT: Medicament; T: Single dose of the herbal cream; NT: Double dose of the herbal cream. Bv: Blood vessel, Cf: Collagen fiber, Ep: Epithelialization, Gt: Granulation tissue, Hf: Hair follicle, Sg: Sweat gland.

Table 7.

Modified histological scoring system adapted by Gupta and Kumar (2015) [44]

Histological parameters	Score
Histological parameters	1	2	3
Epithelialization	Absent	Moderate	Marked
Differentiation	Absent	Present	/
Amount of granulation tissue	Profound	Moderate	Absent
Inflammation	Severe	Moderate	Weak
Collagen fiber orientation	Vertical	Mixed	Horizontal
Angiogenesis	<5/HPF	6-10/HPF	>10/HPF

Table 8.

Mean scores ± SD of histological parameters in all treated groups

Histological parameters	Groups
Histological parameters	C	MDT	VE	T	NT	P value
Epithelialization	1.60 ± 0.54^c	2.60 ± 0.54^ab	1.80 ± 0.44^bc	2.80 ±0.44^a	3.00 ± 0.00^a	0.00
Differentiation	1.40 ± 0.54^a	2.00 ± 0.00^a	1.60 ± 0.54^a	2.00 ± 0.00^a	2.00 ± 0.00^a	0.03
Amount of granulation tissue	2.60 ± 0.54^a	1.20 ± 0.44^b	1.60 ± 0.89^b	3.00 ± 0.00^a	3.00 ± 0.00^a	0.00
Inflammation	1.40 ± 0.54^c	2.20 ± 0.44^b	2.40 ± 0.54^ab	3.00 ± 0.00^a	3.00 ± 0.00^a	0.00
Collagen fiber orientation	1.60 ± 0.54^c	2.40 ± 0.54^ab	2.20 ± 0.44^bc	3.00 ± 0.00^a	3.00 ± 0.00^a	0.00
Angiogenesis	1.60 ± 0.54^c	2.60 ± 0.54^ab	2.00 ± 0.70^bc	3.00 ± 0.00^a	3.00 ± 0.00^a	0.00
Total histological score	10.2 ± 1.09^c	13.00 ± 0.70^b	11.60 ± 1.14^bc	16.80 ± 0.44^a	17.00 ± 0.00^a	0.00

[i] C: Control; MDT: Medicament; VE: Vehicle; T: Single dose of the herbal cream; NT: Double dose of the herbal cream. Groups that do not share the same letter are significantly different.

Straight calibration curve of gallic acid for total phenolic content quantification

Straight calibration curve of quercetin for flavonoid content quantification

Straight calibration curve of ursolic acid for total terpenoids content quantification

Straight calibration curve of ascorbic acid for FRAP assay.

Straight calibration curve of ascorbic acid for TAC assay

DPPH radical scavenging activity Linaria scariosa herbal cream

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DOI: https://doi.org/10.2478/afpuc-2026-0009 | Journal eISSN: 2453-6725 (formerly 1338-6786) | Journal ISSN: 0301-2298

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Language: English

Submitted on: Jan 13, 2026

Accepted on: Aug 6, 2026

Published on: Oct 9, 2026

Published by: Comenius University in Bratislava, Faculty of Pharmacy

In partnership with: Paradigm Publishing Services

Keywords:

Linaria scariosa,

Wound healing,

Herbal cream,

Biological activities

Related subjects:

Pharmacy,

Pharmacy, other

© 2026 Chaima Mouffouk, Soumia Mouffouk, Ahmed Yassine Kalbaza, Aicha Nada Mouffouk, Sara Mouffouk, Abd Elbasset Tamersit, Hachemi Messaoud, Hamada Haba, published by Comenius University in Bratislava, Faculty of Pharmacy
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

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Development of Linaria scariosa Herbal Cream: Formulation, In Vivo Wound Healing, and In Vitro Biological Evaluation

Figures & Tables

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Figure 1.

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Figure 2.

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Table 8.

Figure S1.

Figure S2.

Figure S3.

Figure S4.

Figure S5.

Figure S6.

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