Overexpression of klotho in adipose-derived stem cells protects against UVB-induced photoaging in co-cultured human fibroblasts

Abstract

Co-culture methods are widely used in tissue engineering to drive tissue formation with the direct or indirect interaction of multiple cell types. Klotho is a novel biomarker involved in aging. In this study, we evaluated the protective effects of klotho overexpressed adipose-derived stem cells (ADSCs) against ultraviolet radiation B (UVB)-induced photoaging in co-cultured human skin fibroblasts (HSF2 cell line). Furthermore, the involvement of P38 mitogen-activated protein kinase (MAPK) signaling was investigated. ADSCs were isolated from human subcutaneous adipose tissue and the 3rd generation of ADSCs was used after being identified. Klotho overexpression (OE) lentivirus vectors were constructed and identified in ADSCs. The HSF2 cells were seeded in the upper layer of the Transwell co-culture plate (0.4 µm pore polycarbonate membrane) and ADSCs were seeded in the lower layer. UVB irradiation of HSF2 cells was performed using UVB lamps in uncovered petri dishes at room temperature. The present results indicated that the proliferation of ADSCs was increased by klotho OE. Furthermore the proliferation and collagen content of HSF2 were decreased by UVB irradiation in a dose-dependent manner. By contrast, the protein level of matrix metalloproteinases (MMP) 1, 3 and p-P38 in HSF2 were upregulated. In the co-culture system, relative mRNA expression of MMP-1 and MMP-3 as well as protein level of MMP-1, MMP-3 and p-P38 in HSF2 were reduced by co-culture with klotho overexpressed ADSCs when exposed to UVB (20 mJ/cm2). By contrast, the collagen content of HSF2 was increased. Collectively, OE of klotho in ADSCs notably ameliorates UVB-induced photoaging in co-cultured HSF2, and these effects were potentially achieved by increasing the collagen content and decreasing the protein level of MMP-1, MMP-3 and p-P38.

Application of adipose-derived stem cells in photoaging: basic science and literature review

“…The molecular interaction between fibroblasts and the ECM is obstructed due to the decrease of collagen, which eventually results in the damage of fibroblast function and further collagen decrease [ 94 ]. It is reported that ADSC-Exo can promote the migration ability of HDFs irradiated by UVB [ 95 ], suppress the overexpression of MMP-1 [ 95 , 96 ], MMP-2 [ 95 , 97 ], MMP-3 [ 95 , 96 ], MMP-9 [ 92 , 95 ] and MMP-13 [ 97 ] caused by UVB. Moreover, it can increase collagen I, II, III, and V and elastin expression [ 92 , 95 ].…”

Asam Posfat (H3PO4): Ionic Transformation of Phosphoric Acid in Aqueous Solution

“…Kemungkinan tanah-tanah yang pori-porinya besar, permeabilitasnya mendekati nol (hampir tidak ada aliran), yaitu jika pori-pori tersebut terisolasi (tidak ada hubungan) sesamanya. Permeabilitas juga mungkin mendekati nol apabila (28,(72)(73)(74)(75)(76)(77)(78)(79)(80)(81)(82)(83)(84)(85)(86)(87)(88)(89)(90) pori-pori tanah sangat kecil, seperti pada tanah liat. Sifat dari cairan yang secara langsung berpengaruh terhadap permeabilitas tanah adalah viskositas (viscosity) dan berat jenis (density).…”

Asam Posfat (H3PO4): Ionic Transformation of Phosphoric Acid in Aqueous Solution

“…Metode ini tergolong sederhana dan mudah diaplikasikan. Prinsip yang digunakan identik dengan cara yang dikemukakan De Boodtdan didasarkan pada hukum Darcy (39)(40)(41)(42)(43) .Secara kuantitatif permeabilitas tanah diartikan sebagai kecepatan bergeraknya suatu cairan pada suatu media berpori (28,(72)(73)(74)(75)(76)(77)(78)(79)(80)(81)(82)(83)(84)(85)(86)(87)(88)(89)(90) dalam keadaan jenuh. Dalam hal ini sebagai cairan adalah air, dan sebagai media berpori adalah tanah .…”

“…Kemungkinan tanah-tanah yang poriporinya besar, permeabilitasnya mendekati nol (hampir tidak ada aliran), yaitu jika pori-pori tersebut terisolasi (tidak ada hubungan) sesamanya. Permeabilitas juga mungkin mendekati nol apabila (28,(72)(73)(74)(75)(76)(77)(78)(79)(80)(81)(82)(83)(84)(85)(86)(87)(88)(89)(90) pori-pori tanah sangat kecil, seperti pada tanah liat. Sifat dari cairan yang secara langsung berpengaruh terhadap permeabilitas tanah adalah viskositas (viscosity) dan berat jenis (density).…”