A Randomized Double-Blind, Placebo-Controlled Study to Evaluate the Anti-Skin-Aging Effect of LactoSporin – The Extracellular Metabolite from Bacillus coagulans (Weizmannia coagulans) MTCC 5856 in Healthy Female Volunteers

Figures & data

Figure 1 Participant flow chart at each visit. A flow chart showing the activities carried out on each visit during the study. Open arrows: baseline, grey arrows: Week 5 and black arrows represent week 10.

Table 1 Baseline Demographics

Parameter	Placebo (N=28)	LactoSporin (N=28)	Total (N=56)
Age (Mean ± SD)	43.50 ± 6.4	44.17 ± 6.6	43.83 ± 6.6
Fitzpatrick Skin type (N)
III	3	10	13
IV	11	12	23
V	14	6	20

Note: Demographic details of the study participants.

Abbreviation: SD, Standard deviation.

Figure 2 Consort diagram.

Table 2 Dermatological Assessment of Facial Skin for Signs of Ageing

Assessments	Placebo Formulation			LactoSporin Formulation			^#p-value for Change
	Baseline	Week 5	Week 10	Baseline	Week 5	Week 10
Wrinkles – crow’s feet	5.63±1.49	5.70±1.52	5.74±1.59	6.00±2.01	5.04 ± 1.91*	4.67 ± 1.96*
Change		0.07	0.11		−0.96	−1.33	<0.001
Nasolabial folds	4.98±2.14	5.18±2.1	5.19±2.08	5.09±2.29	4.52±2.18*	4.39±2.18*
Change		0.20	0.21		−0.57	−0.70	<0.001
Frown lines	4.75±1.86	5.02±1.77	5.13±1.83	5.39±2.38	4.61±2.32*	4.38±2.23*
Change		0.27	0.38		−0.79	−1.02	<0.001
Fine lines	7.07±1.25	7.39±1.08	7.4±1.41	7.32±1.82	6.45±1.77*	5.75±1.82*
Change		0.32	0.33		−0.88	−1.57	<0.001
Skin texture	4.93±1.05	4.96±1.07	5.13±1.2	5.07±1.36	4.66±1.42*	4.35±1.48*
Change		0.04	0.20		−0.41	−0.72	<0.001
Skin firmness /elasticity	5.25±1.46	5.38±1.44	5.54±1.49	5.86±1.67	5.45±1.65*	5.15±1.62*
Change		0.13	0.29		−0.41	−0.71	<0.001
Skin hydration	3.29±0.6	3.51±0.62*	3.87±0.51*	3.5±0.64	4.11±0.55*	4.38±0.41*
Change		0.22	0.58		0.61	0.88	<0.001

Notes: Dermatological assessments were carried out at baseline, week 5 and week 10. The values in photo numeric scale are represented as Mean±SD and the change in the photo numerical scale readings from baseline to week 5 and week 10 are presented. For crow’s feet, fine lines, frown lines, nasolabial folds, skin texture, and firmness, the scale was from 0 to 9 and lower mean values indicate better condition. An increase in skin hydration is measured as the higher mean value. The ^#P value was computed for the change in values from baseline and week 10 in LactoSporin and Placebo groups. *P<0.05 between baseline and week 5 and week 10 within the group.

Abbreviation: PNS, Photo Numeric Scale.

Table 3 Instrument Analysis of Facial Skin for Signs of Ageing

Assessments	Placebo Formulation			LactoSporin Formulation			^#p-value for Change
	Baseline	Week 5	Week 10	Baseline	Week 5	Week 10
Forehead wrinkles (mm)	18.42 ± 5.3	17.71 ± 5.3*	17.34 ± 5.2*	16.44 ± 4.9	15.10 ± 4.8*	14.58 ± 4.7*
Change		−0.7	−1.07		−1.34	−1.86	0.022
Forehead texture (µm)	13.40 ± 2.8	12.69 ± 2.7*	12.23 ± 2.7*	13.45 ± 2.4	12.61 ± 2.1*	11.91 ± 2.1*
Change		−0.71	−1.17		−0.84	−1.54	0.049
Crow’s feet (mm)	17.71 ± 4.8	17.32 ± 4.8*	16.79 ± 4.7*	17.03 ± 3.9	16.33 ± 3.9*	15.61 ± 3.8*
Change		−0.39	−0.92		−0.7	−1.42	0.042
Nasolabial folds (mm)	15.66 ± 3.8	15.00 ± 3.9*	14.49 ± 3.7*	14.84 ± 4.0	14.06 ± 3.8*	13.36 ± 3.8*
Change		−0.65	−1.17		−0.77	−1.47	0.047
Under eye-fine lines (mm)	12.58 ± 2.7	12.09 ± 2.6*	11.54 ± 2.5*	12.26 ± 2.2	11.66 ± 2.1*	10.90 ± 2.1*
Change		−0.49	−1.04		−0.6	−1.36	0.078
Skin hydration (AU)	35.91 ± 2.5	41.53 ± 2.4*	47.05 ± 2.5*	36.56 ± 2.2	41.94 ± 2.5*	47.36 ± 2.3*
Change		5.61	11.13		5.38	10.8	0.15
Transepidermal water loss (g/m^2h)	10.26 ± 1.6	9.57 ± 1.5*	9.04 ± 1.6*	10.05 ± 2.6	9.42 ± 2.6*	8.85 ± 2.5*
Change		−0.69	−1.22		−0.63	−1.2	0.52
R2 – Skin elasticity (mm)	0.58 ± 0.3	0.68 ± 0.1*	0.74 ± 0. 2*	0.55 ± 0.1	0.66 ± 0.1*	0.72 ± 0.1*
Change		0.09	0.16		0.1	0.17	0.73
R5 – Skin elasticity (mm)	0.64 ± 0.2	0.71 ± 0.2*	0.76 ± 0.2*	0.58 ± 0.2	0.68 ± 0.1*	0.67 ± 0.1*
Change		0.07	0.12		0.1	0.09	0.39
R7 – Skin elasticity (mm)	0.41± 0.1	0.47 ± 0.1*	0.54 ± 0.1*	0.37 ± 0.1	0.44 ± 0.1*	0.48 ± 0.1*
Change		0.06	0.13		0.07	0.11	0.34

Notes: The values for instrument-based assessments carried out at baseline, week 5 and week 10 are represented at Mean ± SD, and the change from baseline to week 5 and week 10 is presented. The ^#P value represents the difference between LactoSporin and Placebo groups at week 10. Forehead wrinkles, texture, nasolabial folds, crow’s feet, and under eye fine lines were assessed by Antera- 3D, Skin hydration was by Corneometer, barrier function by Tewameter and elasticity by Cutometer. *P<0.05 between baseline and end of the study within the group.

Abbreviation: AU, arbitrary units.

Figure 3 Analysis of topographical features of the skin by Antera 3D. Representative images from Antera 3D, showing the changes in crow’s feet, under-eye fine lines, nasolabial folds, and forehead fine lines in placebo and LactoSporin at baseline, week 5 and week 10. The blue marks represent wrinkles within the area of interest. A lower value indicates less severity and an improvement.

Figure 4 VISIA representative images. Representative VISIA images of two participants each from A -LactoSporin and B- Placebo group taken at baseline, week 5 and week 10. Full face digital images were taken using VISIA-CR Blue 4.3 Camera Imaging System on day 0, day 35 and at day 70 for all the participants. The representative VISIA images of two participants each from LactoSporin (A)and Placebo (B) taken at day 01, day 35 and day 70 are depicted.

Table 4 Participant Response to Self-Assessment Questionnaire

Questions	Placebo Formulation		LactoSporin Formulation
	Week 5	Week 10	Week 5	Week 10
Does your skin look firmer	17 (60.71%)	26 (92.86%)	19 (67.86%)	27 (96.43%)
Does your skin look free of fine lines/wrinkles on your face?	10 (35.71%)	26 (92.86%)	16 (57.14%)	27 (96.43%)
The test products make my skin moisturized.	16 (57.14%)	25 (89.29%)	18 (64.29%)	27 (96.43%)

Notes: Response recorded by participants on their perception of the skin condition. The number and percentage of participants who agreed (4 or 5 in the response scale) that they felt an improvement in skin condition is represented as number and percentage.