LOXL2 from human amniotic mesenchymal stem cells accelerates wound epithelialization by promoting differentiation and migration of keratinocytes

Abstract

In this study, we identified wound healing-related proteins secreted by human amniotic epithelial cells (hAECs) and human amniotic mesenchymal stem cells (hAMSCs). We observed increased migration and reduced proliferation and differentiation when keratinocytes were co-cultured in media conditioned by hAECs (hAECs-CM) and hAMSCs (hAMSCs-CM). Label-free mass spectrometry and bioinformatic analyses of the hAECs-CM and hAMSCs-CM proteome revealed several proteins associated with wound healing, angiogenesis, cellular differentiation, immune response and cell motility. The levels of the proteins related to wound healing, including CTHRC1, LOXL2 and LGALS1, were significantly higher in hAMSCs-CM than hAECs-CM. LOXL2 significantly enhanced in vitro keratinocyte migration and differentiation compared to CTHRC1 and LGALS1. Moreover, LOXL2 enhanced keratinocyte migration and differentiation by activating the JNK signaling pathway. We observed significant reduction in the in vitro migration and differentiation of keratinocytes when co-cultured with medium conditioned by LOXL2-silenced hAMSCs and when treated with 10 μM SP600125, a specific JNK inhibitor. Treatment with hAMSCs-CM and LOXL2 significantly accelerated wound healing in the murine skin wound model. These findings show that LOXL2 promotes wound healing by inducing keratinocyte migration and differentiation via a JNK signaling pathway.

Systematic comparation of the biological and transcriptomic landscapes of human amniotic mesenchymal stem cells under serum-containing and serum-free conditions

“…Therefore, hAMSC-based therapy has been widely explored to promote endogenous regeneration and alleviate the untoward effect of traditional treatment [ 40 ]. To date, hAMSCs have been reported as appropriate sources with therapeutic effects in regenerative medicine and tissue engineering such as premature ovarian insufficiency (POI) [ 41 ], premature ovarian failure (POF) [ 36 ], wound healing and epithelialization [ 42 ], hepatocellular carcinoma [ 43 ], and endogenous bone regeneration [ 40 ]. Generally, hAMSCs function via direct- and trans-differentiation, paracrine, and autocrine (e.g., extracellular proteins, anti-bacterial peptides, growth factors, angio-modulatory cytokines, bioactive factors), immunoregulation (e.g., anti-inflammatory agents, polarization of T lymphocytes, or macrophages), activating endogenous regeneration and serving as constitutive microenvironment [ 17 , 39 – 41 ].…”

Characteristics and Therapeutic Potential of Human Amnion-Derived Stem Cells

“…Yu et al establish a new model for reconstruction of bilayer TE skin with hAMSCs and hAESCs [ 177 ] and the TE skin was similar in morphology to human skin, in which they had stratified epidermis and underlying dermis and successfully repaired full thickness skin defects [ 178 ]. Although both hAMSC-CM and hAESC-CM have been proved to promote wound healing, the levels of wound healing related proteins such as CTHRC1, LOXL2, and LGALS1 in hAMSC-CM were significantly higher than those in hAESC-CM [ 179 ].…”

Mesenchymal Stromal Cell-Conditioned Medium for Skin Diseases: A Systematic Review

“…Thirty-one studies evaluated the potential of MSC-CM for treating non-diabetic wounds (Table 1). To obtain CM, cells were mainly isolated from human tissues (74.19%, 23/31): from adipose tissue (Cho et al, 2010;Heo et al, 2011;Deng et al, 2017;Yuan et al, 2018;Chen et al, 2021) (21.74%, 5/23), amnion (Yoon et al, 2010;Jun et al, 2014;Park et al, 2018;He et al, 2020) (17.34%, 4/23), umbilical cord blood (Lee et al, 2011;Dong et al, 2017;Raj et al, 2019;Rong et al, 2019) (17.34%, 4/23), bone marrow (Tamari et al, 2013;Chen et al, 2014;Ahangar et al, 2020) (13.04%, 3/23), and Wharton's jelly (Fong et al, 2014;Tam et al, 2014;Sabzevari et al, 2020) (13.04%, 3/23). Human dental pulp (Zhou et al, 2020) and skin (Robert et al, 2019) were also used.…”

“…Regarding the route of administration, MSC-CM was mainly used topically (51.61%, 16/31), applied to the wounds or around them in cream, hydrogel, or membranes (Heo et al, 2011;Lee et al, 2011;Chen et al, 2014;Jun et al, 2014;Sun et al, 2014;Mehanna et al, 2015;Park et al, 2018;Joseph et al, 2020). MSC-CM was also injected (41.94%, 13/31), mainly subcutaneously (25.81%, 8/31) (Templin et al, 2009;Lee et al, 2011;Deng et al, 2017;He et al, 2020;Zhou et al, 2020) but also intradermally (Cho et al, 2010) and intraperitoneally (Fong et al, 2014). Subcutaneously injected and topically applied concomitant MSC-CM was also tested (Chen et al, 2008;Yoon et al, 2010).…”