Ability of sodium copper chlorophyllin complex to repair photoaged skin by stimulation of biomarkers in human extracellular matrix

Figures & data

Figure 1 Procollagen 1 dermal histology.

Notes: Immunohistochemical staining for procollagen I, showing an increased level of staining in both the test material (A) (subject 4_site 1_procollagen a) and reference control (B) (subject 4_site 2_procollagen b), when compared to negative control (C) (subject 4_site 3_procollagen c). Scale bar =200 μm.

Figure 2 Fibrillin 1 dermal histology.

Notes: Immunohistochemical staining for amyloid P (fibrillin surrogate marker), showing increased fibrillin signal in both the test material (A) (subject 4_site1_fibrillin_amyp a), and reference control (B) (subject4_site2_fibrillin_amyp b), when compared to the negative control (C) (subject4_site3_fibrillin_amyp c). Scale bar =200 μm.

Figure 3 Epidermal and dermal mucin histology.

Notes: Histochemical staining for mucin via colloidal iron to highlight both epidermal and dermal mucins. An increased amount of mucin is seen in both compartments in both the test (A) (subject4_site1_colfe_orig a) and reference material (B) (subject4_site2_colfe_orig b) compared to the negative control (C) (subject4_site3_colfe_orig c). Scale bar =200 μm.

Table 1 Subject disposition and demographics

	All subjects (N=5)	Biopsy subjects (N=4)
Age (years)
Mean (SD)	50 (3.9)	51 (3.7)
Median (min, max)	49 (46, 55)	51 (47, 55)
Sex, n (%)
Female	5 (100)	4 (100)
Ethnicity/race, n (%)
Caucasian	5 (100)	4 (100)
Fitzpatrick skin type, n (%)
II	4 (80)	3 (75)
III	1 (20)	1 (25)

Abbreviations: SD, standard deviation; min, minimum; max, maximum.

Table 2 Descriptive statistics for biopsy results at day 12 (N=4 subjects)

Parameter Treatment	Mean	Standard deviation	Minimum	Median	Maximum
Procollagen I
Test material	1.25	0.50	1.00	1.00	2.00
Reference control	1.50	0.58	1.00	1.50	2.00
Negative control	1.00	0.00	1.00	1.00	1.00
Fibrillin/amyloid P
Test material	1.25	0.50	1.00	1.00	2.00
Reference control	1.50	0.58	1.00	1.50	2.00
Negative control	0.25	0.50	0.00	0.00	1.00
Epidermal mucins (colloidal iron)
Test material	1.25	0.50	1.00	1.00	2.00
Reference control	1.50	0.58	1.00	1.50	2.00
Negative control	0.50	0.58	0.00	0.50	1.00
Dermal mucin
Test material	1.75	0.96	1.00	1.50	3.00
Reference control	1.25	0.50	1.00	1.00	2.00
Negative control	1.00	0.00	1.00	1.00	1.00

Notes: Fibrillin, epidermal and dermal mucin are rated on four point (0–3) scale: 0 =normal, 3 =marked increased staining. Procollagen is rated on a 5 point (1–5) scale: 1 =<5% cells positive, 5 =>30% cells positive.

Table 3 Comparison details between test products for biopsies at day 12

Parameter	Overall P-valuea	Comparison	Estimated mean difference	Standard error	Pairwise P-valuea
Procollagen I	0.244	Test material vs reference control	−0.25	0.26	0.632
		Test material vs negative control	0.25	0.26	0.632
		Reference control vs negative control	0.50	0.26	0.220
Fibrillin/amyloid P	0.002	Test material vs reference control	−0.25	0.20	0.483
		Test material vs negative control	1.00	0.20	0.006
		Reference control vs negative control	1.25	0.20	0.002
Epidermal mucins (Colloidal iron or Alcian blue/hyaluronidase)	0.007	Test material vs reference control	−0.25	0.20	0.483
		Test material vs negative control	0.75	0.20	0.024
		Reference control vs negative control	1.00	0.20	0.006
Dermal mucin (Colloidal iron or Alcian blue/hyaluronidase)	0.178	Test material vs reference control	0.50	0.35	0.392
		Test material vs negative control	0.75	0.35	0.165
		Reference control vs negative control	0.25	0.35	0.768

Notes:

a P-value was calculated using a mixed effect model where test product is a fixed effect and subject is a random effect. Tukey’s honestly significant difference test was used for pairwise treatment comparison. The null hypothesis is that the mean scores are equal between treatments.