102 SKIN CARE
concentration, indicating the success in scale- up fermentation. Besides, the test results of final products, including SDS-PAGE, Western Blot, MALDI- TOF MS, FT-IR, were the same as we carried in the laboratory fermentation and the amino acid sequence of SRHC Type XVII was totally the same as we designed after verification by amino acid sequencing. Collectively, we have realized scale-up
fermentation of recombinant collagen type XVII with secretory strategy and the homology degree with human native type XVII collagen is >90%.
Efficacy test of SRHC Type XVII as a new raw material for cosmetics We carried some efficacy test to evaluate whether SRHC Type XVII could be used as a new raw material for cosmetics. We first carried some experiments regarding cell bioactivity including cell migration activity and cell adhesion activity. As we expected, after rational design and
careful selection of amino acid sequence on collagen type XVII, SRHC Type XVII exhibited even greater bioactivity than human collagen in promoting cell migration and adhesion (Figure 3A). On the transcriptional level, we observed
significant increase in the expression level of Aquaporin 3-encoding gene and decrease in the expression level of matrix metalloproteinase MMP3 and MMP9 encoding genes, which would be helpful in moisturizing and collagen maintenance respectively (Figure 3B).
After treatment with 0.1% SRHC Type XVII,
we could observe significant increase in the expression level of basement membrane- related proteins, including collagen type IV, type VII, type XVII, ITGβ4, ITGα6, Lamin and Nidogen (Fig.3C), indicating that SRHC Type XVII would have a great efficacy on repairing damaged DEJ. So, we adopted an ex vivo skin tissue
model to confirm this approximation. We used the immunofluorescence staining technology and found that the treatment of 0.1% SRHC Type XVII could greatly increase the expression level of human collagen type XVII (Figure 3D). In addition, after UV damage, there was
break in the basement membrane but the treatment of 0.1% SRHC Type XVII could repair DEJ damage and the DEJ was intact (Figure 3E).
Conclusion We are the first to achieve scale fermentation of recombinant human collagen type XVII (trade name: SRHC Type XVII) around the world and evaluate the efficacy of SRHC Type XVII as a new raw material for cosmetics. We found that the treatment of SRHC
Type XVII can (1): promote cell migration and adhesion; (2) increase the expression level of fundamental proteins involved in basement membrane; and (3) repair dermal–epidermal junction (DEJ) damage and significantly increase the endogenous expression level of collagen type XVII both at the transcriptional and translational level.
PERSONAL CARE March 2024
Figure 3: Efficacy test of SRHC Type XVII as raw materials for cosmetics. A: Cell migration and adhesion activity of NIH/3T3 cells. B: Expression levels of AQP3 gene in HaCat cells, MMP3 and MMP9 genes in HFF-1 cells normalized to GAPDH quantified by qPCR. C: Expression levels of basement-related genes, including collagen type IV, type VII, type XVII, ITGβ4, ITGα6, lamin and Nidogen normalized to GAPDH quantified by qPCR. D: Immunofluorescence staing results of collagen type XVII expressed in the HaCat cells on the basement membrane. E: Transmission electron microscope (TEM) images of dermal-epidermal junction
Taken together, SRHC Type XVII can be used as a novel and attractive raw material of cosmetics.
PC
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www.personalcaremagazine.com D A Cell migration activity of NIH/3T3
100 80 60 40 20 0
24 h B
2.0 1.5 1.0 0.5 0.0
* 48 h Without treatment ■ Treatment with SRHC Type XVII ■ 3 ***
1.5 1.0 0.5 0.0
AQP3 * 2 1 0 MMP9 ** ** *** *** *** * C Without treatment ■ Treatment with SRHC Type XVII ■ 1.5 1.0 0.5 0.0 Cell adhesion activity of NIH/3T3
Blank control
Negative control
E
Blank control
Negative control
Positive control
0.1% SRHC Type XVII
Positive control
0.1% SRHC Type XVII
Relative Gene Expression (a.u.) Normalized to GAPDH
Migration activity x100% (a.u.)
Relative Gene Expression (a.u.) Normalized to GAPDH
OD492
Relative Gene Expression (a.u.) Normalized to GAPDH
(a.u.)
Collagen type IV
Collagen type VII Collagen type XVII
ITGβ4 ITGα6
Lamin Nidogen
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