Mécano-Stimulation™ of the skin improves sagging score and induces beneficial functional modification of the fibroblasts: clinical, biological, and histological evaluations

Figures & data

Figure 1 Ezure scoring scale.

Notes: Skin sagging is evaluated through the Ezure scoring scale:¹¹ facial cutaneous sagging is graded from 0 to 5 on three areas of the hemiface (A), upper cheek (B), lower cheek (C), and lateral cheek (D). Arrows and arrowheads point out specific areas of interest defining skin sagging stages. Volunteers selected in the study were of grade 1–3.

Figure 2 Double stimulation.

Notes: The Mécano-Stimulation™ technique consists of two motorized flaps located inside a treatment chamber and dedicated to the treatment of the face or small areas. The treatment forces combine sequential aspiration and beating flaps creating, respectively, vertical and horizontal stimulation. Settings allow obtaining different intensity and rhythmicity of the aspiration or beating frequency of the flaps. Lift heads used in this study were “motorized”: heads contain a small engine allowing the flaps to beat in a regular and permanent way, synchronized specifically with the sequential aspiration.

Table 1 Results of the skin sagging scores

Zones	Number of volunteers with improved score (out of 30 subjects)		P
	Treated hemiface	Nontreated hemiface
B	10 (33%)	8 (27%)	0.573
C	7 (57%)	11 (37%)	0.120
D	13 (43%)	8 (27%)	0.176
At least one among three	22 (73%)	16 (53%)	0.108
At least C or D	21 (70%)	15 (50%)	0.114

Notes: Number of volunteers with improved clinical sagging score (Ezure scale¹¹) is presented. A clinical improvement of skin sagging of at least one parameter of the score was observed in 73% (n=22) of the subjects on the treated side. B zone is the upper part of the cheek/nasolabial folds, C zone is the lower part of the cheeks, and D zone is the lateral part of the cheek. Clinical improvement of the sagging of at least C or D areas (where the massage was the more efficient) was observed in 70% of treated hemiface of the subjects.

Table 2 Results for CLBT™ scoring before (T0) and after (T1) the 24 treatment sessions

Score (Musnier et al^Citation12)	Treated hemiface		Nontreated hemiface
	T0 (mean ± SD)	T1 (mean ± SD)	T0 (mean ± SD)	T1 (mean ± SD)
Color score (the lower the score the more homogeneous the skin is)	12.60±3.10	10.90±2.98*	12.03±2.89	11.83±2.89§
Pink red	40.33±19.56	33.00±18.03*	40.00±17.42	38.33±16.42§
Olive	36.67±10.93	24.67±9.00*	36.00±10.37	32.67±9.07§
Beige	54.00±12.76	45.00±13.06*	53.33±13.73	50.67±13.67§
Light pink	69.67±12.17	59.00±17.09*	69.67±12.99	66.33±13.2§
Transparency	3.56±1.93	4.57±1.50*	3.55±1.84	3.59±1.45§
Brightness	2.43±1.88	3.04±1.64*	2.44±1.80	2.57±1.53§
Luminosity	2.08±1.12	4.11±1.69*	2.01±1.06	2.94±1.38*,§

Notes: The visual analysis scale (CLBT scoring¹²) follows seven descriptors that evaluate four different colors (scale between 10% and 100%) and three parameters of skin transparency, brightness, and luminosity (0–10 scale). Compared to baseline, all scores were significantly improved after treatment on the treated hemiface (P<0.05); at T1, the color scores as well as skin transparency, brightness, and luminosity were significantly improved on the treated compared to the nontreated hemiface. All the parameters presented a significant improvement in terms of time/treatment evolution.

* Significantly different from T0 within each hemiface.

§ Significantly different from treated hemiface at T1.

Abbreviations: CLBT, Color, Luminosity, Brightness and Transparency Scoring; SD, standard deviation; T, time.

Figure 3 CLBT™ scoring before (T0) and after (T1) the 24 treatment sessions: results for the color components.

Notes: The visual analysis scale (CLBT scoring¹²) follows seven descriptors that evaluate between 10% and 100% of four different colors: pink red (A), olive (B), beige (C), and light pink (D). All these color parameters presented a significant improvement (decrease) in terms of time/treatment evolution, in favor of the treated (blue) versus nontreated (red) side. *P<0.05.

Abbreviations: CLBT, Color, Luminosity, Brightness and Transparency Scoring; T, time.

Figure 4 CLBT™ scoring before (T0) and after (T1) the 24 treatment sessions: results for transparency, brightness, and luminosity.

Notes: The visual analysis scale (CLBT scoring¹²) evaluates three parameters of skin transparency (A), brightness (B), and luminosity (C) on a 0–10 scale; a significant improvement (increase) in terms of time/treatment evolution is pictured in all cases, in favor of the treated (blue) versus nontreated (red) hemiface. *P<0.05.
Abbreviations: CLBT, Color, Luminosity, Brightness and Transparency Scoring; T, time.

Table 3 Clinical scoring of skin radiance: evaluation through relief before (T0) and after (T1) the 24 treatment sessions

Score	Treated hemiface		Nontreated hemiface
	T0 (mean ± SD)	T1 (mean ± SD)	T0 (mean ± SD)	T1 (mean ± SD)
Surface irregularity	5.13±1.20	4.07±1.05*	5.13±1.20	5.00±1.05§
Fine lines, wrinkles, and blemishes	4.73±1.26	3.73±1.05*	4.80±1.24	4.77±1.04§
Skin texture	3.73±1.84	3.17±1.88 *	3.77±1.87	3.53±2.03§
Under-eye dark circles	4.63±1.71	3.97±1.85*	4.77±1.72	4.57±1.74§
Buttons/microcysts	1.23±1.07	0.80±0.85*	1.17±1.02	0.70±0.79§
Relief score	19.47±3.85	15.73±4.02*	19.63±3.74	18.57±3.41§

Notes: An analogical scale (from 0 to 9) was used to score the radiance through relief parameters, before and after the treatment: surface irregularity, fine lines/wrinkles/blemishes, skin pores, under-eye dark circles, and buttons/microcysts. A global relief score was defined as the sum of the previous scores (the lower the score, the better the relief is). A significant improvement in terms of time/treatment evolution was observed for all the evaluated parameters.

* Significantly different from T0 within each hemiface.

§ Significantly different from treated hemiface at T1.

Abbreviations: SD, standard deviation; T, time.

Figure 5 Clinical scoring of skin radiance: relief surface irregularity.

Notes: An analogical scale (from 0 to 9) was used to score the radiance through relief parameters: among those, surface irregularity was evaluated from small (0) to large (9) before (T0) and after (T1) the treatment. The figure represents a significant improvement (decrease) in terms of time/treatment evolution observed; surface irregularity score was improved by 20.7% on treated side (blue) against 2.5% on nontreated side (red). *P<0.05.
Abbreviation: T, time.

Table 4 Clinical scoring of skin radiance: evaluation through skin color before (T0) and after (T1) the 24 treatment sessions

Score	Treated hemiface		Nontreated hemiface
	T0 (mean ± SD)	T1 (mean ± SD)	T0 (mean ± SD)	T1 (mean ± SD)
Skin complexion heterogeneity	3.40±0.86	2.93±0.98*	3.23±0.73	3.27±0.87§
Under-eye dark circles	3.57±1.52	3.03±1.50*	3.53±1.66	3.47±1.59§
Redness/rosacea	2.87±1.59	2.60±1.45	2.80±1.52	2.87±1.36
Skin spots (hyperpigmentation)	2.03±1.19	1.97±1.22	1.97±1.35	1.93±1.34
Buttons/scars	0.73±0.87	0.37±0.49*	0.50±0.73	0.30±0.53
Color score	12.60±3.10	10.90±2.98*	12.03±2.89	11.83±2.89

Notes: The scoring of the radiance was also assessed through color parameters with an analogical scale (0–9): skin complexion heterogeneity, under-eye dark circles, redness/rosacea, skin spots, and buttons/scars. A global color score was defined as the sum of the previous scores (the lower the score, the better the radiance is). Results are presented before (T0) and after (T1) the 24 sessions of treatment. A significant improvement in terms of time/treatment evolution was observed for skin complexion heterogeneity, under-eye dark circles, redness, and color score.

* Significantly different from T0 within each hemiface.

§ Significantly different from treated hemiface at T1.

Abbreviation: T, time.

Figure 6 Clinical scoring of skin radiance: color skin complexion heterogeneity.

Notes: The scoring of the radiance was also assessed through color parameters: skin complexion heterogeneity score was evaluated with an analogical scale (from 0= homogeneous to 9= heterogeneous) and is presented before (T0) and after (T1) the 24 sessions of treatment. At T1, the score was significantly improved (decrease) on the treated side (blue) compared to the nontreated hemiface (red); a significant improvement in terms of time/treatment evolution was observed. Skin complexion heterogeneity score was improved by 13.8% on the treated side while a worsening was noticed on the nontreated side (+1.2%). *P<0.05.
Abbreviation: T, time.

Table 5 Results of self-assessment evaluation

	Improvement on treated side at T1 compared to T0 (vs untreated side)	Significant difference at T1 between treated and untreated	Significant difference in evolution between the treated and untreated
Wrinkles	13.6% (4.8%)	Yes (P=0.004)
Roughness	3.8% (−5.1%)	Yes (P=0.028)	Yes (P=0.016)
Fine lines	−3.1% (−10.5%)
Nasolabial fold	21.3% (11.6%)	Yes (P=0.089)
Elasticity	11.2% (0.5%)	Yes (P=0.003)
Firmness	23.1% (2.5%)	Yes (P=0.003)	Yes (P=0.001)
Radiance	26.3% (14%)	Yes (P=0.011)	Yes (P=0.026)
Softness	2.1% (−7.4%)	Yes (P=0.018)	Yes (P=0.016)
Under-eye dark circles	18.8% (0%)	Yes (P=0.001)

Notes: At T0 and T1, volunteers were asked to score some parameters which characterized their skin (described as presence/absence and the intensity of each parameter). Self-assessment showed improvement on treated side for nine of the ten parameters evaluated; in all cases, nontreated side was felt as less improved or even to have worsened. Difference at T1 between treated and untreated sides were significant for seven of the ten parameters, and trend for one. Significant evolution of treated side was noticed for roughness, firmness, radiance, and softness, compared to untreated hemiface.

Abbreviation: T, time.

Table 6 Self-assessment of subjects skin at T1 (end of the treatment) and T2 (1 month after the end of the treatment), compared to T0

		% of subjects with positive answer
		Treated	Not treated
Firmer skin	T1	70	23
	T2	75	37
More elastic skin	T1	73	20
	T2	75	24
Smoother skin	T1	67	23
	T2	87	12
Less wrinkled skin	T1	87	10
	T2	75	12
Less relaxed skin	T1	77	20
	T2	75	25
Brighter skin	T1	20	53
	T2	62	24

Notes: The result of the self-assessment questionnaire allowing the volunteers to rate their skin state at T1 and T2, compared to T0 is shown. The proportion of positive answers for each item is presented; the results demonstrated a strong effect on the treated side compared with the nontreated side, for all parameters, except brightness at T1. After treatment, patients found their skin firmer, smoother, less wrinkled and less sagged, with a remanent effect 1 month after the last treatment.

Abbreviation: T, time.

Figure 7 Skin sagging evolution during the study: examples on two volunteers treated on the left side of the face before and after 24 sessions of treatment.

Notes: Standardized photographs illustrated an improvement of the face oval: Two examples are pictured: before treatment (A, C) where skin sagging is indicated with arrows on lateral and lower cheek; after 24 sessions of treatment, and (B, D) skin slackening is reduced.

Figure 8 Effect of Mécano-Stimulation™ on fibroblast retraction according to time.

Notes: Fibroblasts capacity to retract free-floating collagen lattices reflects their migration, and is presented to time. The retraction of collagen lattices obtained with fibroblasts exposed to Mécano-Stimulation™ was significantly increased (decreasing of the diameter of retracted collagen lattices) compared to nonstimulated fibroblasts (P<0.001).

Figure 9 Quantification of contractile forces developed by fibroblasts after the action of Mécano-Stimulation™ using GlaSbox^® device.

Notes: The GlaSbox^® device contains eight rectangular culture wells in which lattices have been developed; two opposite silicon beams with strain gauges allow force measurements to be obtained. The results are expressed as contractile forces (arbitrary unit) developed by fibroblasts subjected or not subjected to Mécano-Stimulation™ according to the time. No significant difference of the development of contractile forces was observed between Mécano-Stimulation group and control group.

Figure 10 Collagen I, elastin, and hyaluronic acid synthesis of fibroblasts with and without the action of Mécano-Stimulation™.

Notes: The quantification of collagen I and hyaluronic acid was evaluated using an enzyme-linked immunosorbent assay kit. The concentration in the samples was determined by comparing the optical density of the samples to the standard curve. The total soluble elastin was assayed by a colorimetric method (Fastin Elastin assay). Results are expressed as % respective control. At T1, nonsignificant increase in the synthesis of collagen I (A) was observed between the treated and untreated hemiface (107.8%±69.8% and 100.0%±0.0%/control). At T1, an increase in the synthesis of elastin (B) and hyaluronic acid (C) was observed between the treated and untreated hemiface (P=0.175 and P=0.268, respectively).
Abbreviation: T, time.

Figure 11 MMP-1, MMP-9, and TIMP-1 synthesis of fibroblasts with and without the action of Mécano-Stimulation™.

Notes: The quantification of MMP-1, MMP-9, and TIMP-1 was obtained using an enzyme-linked immunosorbent assay kit. Results are expressed as % respective control. No significant change was observed in the synthesis of MMP-1 (A) and TIMP-1 (C). At T1, a significant increase in the synthesis of MMP-9 (B) was observed between the treated and untreated hemiface (P=0.033). *P<0.05 versus control.
Abbreviations: MMP, matrix metalloproteinase; TIMP, tissue inhibitor of metalloproteinases; T, time.

Figure 12 Examples of ultrastructure observed when comparing the treated versus untreated side.

Notes: Examples of ultrastructure of the papillary dermis observed when comparing the treated versus untreated side. Untreated side (A): Presence of an amorphous substance (*) in the spaces between fragmented EFs. Treated side (B, C, D, and E). (B) Restructured EFs in the vicinity of Fb with clear cytoplasm and developed mitochondria. Fb express well-developed rough endoplasmic network (C) and plasma membrane–attached small secretory vesicles often facing the EFs (D). (E) Immunogold labeling of α-smooth muscle actin in the Fb of the papillary dermis.
Abbreviations: EFs, elastic fibers; Fb, fibroblasts.

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