Mesenchymal stem cells combined with autocrosslinked hyaluronic acid improve mouse ovarian function by activating the PI3K-AKT pathway in a paracrine manner

Abstract

Background Declining ovarian function in advance-aged women and in premature ovarian insufficiency (POI) patients seriously affects quality of life, and there is currently no effective treatment to rescue ovarian function in clinic. Stem cell transplantation is a promising therapeutic strategy for ovarian aging, but its clinical application is limited due to the low efficiency and unclear mechanism. Here, a novel combination of umbilical cord-mesenchymal stem cells (UC-MSCs) and autocrosslinked hyaluronic acid (HA) gel is explored to rescue ovarian reserve and fecundity in POI and naturally aging mice. Methods To investigate HA prolonged the survival after UC-MSCs transplantation, PCR and immunofluorescence were performed to track the cells on day 1, 3, 7 and 14 after transplantation. The effects of HA on UC-MSCs were analyzed by CCK8 assay, RNA-sequencing and 440 cytokine array. In vivo experiments were conducted to evaluate the therapeutic effects of UC-MSCs combined with HA transplantation in 4-vinylcyclohexene diepoxide (VCD)-induced POI mice and naturally aging mice model. Ovarian function was analyzed by ovarian morphology, follicle counts, estrous cycle, hormone levels and fertility ability. To investigate the mechanisms of stem cell therapy, conditioned medium was collected from UC-MSCs and fibroblast. Both in vitro ovarian culture model and 440 cytokine array were applied to assess the paracrine effect and determine the underlying mechanism. Hepatocyte growth factor (HGF) was identified as an effective factor and verified by HGF cytokine/neutralization antibody supplementation into ovarian culture system. Results HA not only prolongs the retention of UC-MSCs in the ovary, but also boosts their secretory function, and UC-MSCs promote follicular survival by activating the PI3K-AKT pathway through a paracrine mechanism both in vitro and in vivo. More importantly, HGF is identified as the key functional cytokine secreted by MSCs. Conclusions The results show that HA is an excellent cell scaffold to improve the treatment efficiency of UC-MSCs for ovarian aging under both physiological and pathological conditions, and the therapeutic mechanism is through activation of the PI3K-AKT pathway via HGF. These findings will facilitate the clinical application of MSCs transplantation for ovarian disorders.

Human Umbilical Cord-Based Therapeutics: Stem Cells and Blood Derivatives for Female Reproductive Medicine

“…Notably, only Sun's group [200] has translated the use of collagen scaffolds from mice [201] to humans. Nevertheless, in mice, both hyaluronic acid and collagen scaffolds sustained the effects of the hUC-MSCs, improving ovarian morphology, cell proliferation, angiogenesis, and ultimately, restoring fertility [197,198,201]. Likewise, women treated with the hUC-MSC-collagen complex achieved a complete restoration of fertility, with primordial follicle activation (determined by the expression of phosphorylation of Forkhead box O3a (FOXO3a) and Forkhead box O1 (FOXO 1)), increased proportion of growing follicles, and consequent high serum estradiol concentration [200].…”

“…While hyaluronic acid is the most popular scaffold used to develop alternative treatments in murine models of ovarian pathologies [197,198], other synthetic scaffolds have been mixed with biological products to enhance their pharmacodynamics [199]. Notably, only Sun's group [200] has translated the use of collagen scaffolds from mice [201] to humans.…”

Biomaterials and advanced technologies for the evaluation and treatment of ovarian aging

“… Or Friedman [ 227 ] 2012 HA—rich biogel Immunodeficient mice Improved ovarian graft survival. Wenlin Jiao [ 228 ] 2022 A combination of UCMSCs and HA gel 4-Vinylcyclohexene diepoxide -induced POI mouse model Improved follicular survival. Eun-Young Shin [ 229 ] 2021 HA gel scaffolder Cisplatin-induced POI mouse model Restored the ovarian structure and function and improved the quality of oocyte and embryo as well as the regularity of estrus cycle.…”

“…Self-linked HA is a good cell scaffold to improve the transplantation of stem cells in the treatment of ovarian aging. Jiao et al explored a combination of hUCMSCs and HA gel to rescue ovarian reserve and fecundity in a POI mouse model [ 228 ]. The HA gel not only increased the local retention of stem cells in the ovary, but also enhanced the paracrine function of hUCMSCs.…”

Cell-Free Fat Extract Improves Ovarian Function and Fertility in Mice With Advanced Age

“…For example, adipose-derived stem cells can rescue the ovarian function via secretion of HGF and bFGF from stem cells ( 16 , 17 ). Many growth factors were vital for maintaining the balance of ovarian microenvironment and further affecting the functions of ovary, however, the low of cytokines production and expensive treatment costs are not conducive to their wide application ( 18 , 19 ). Despite the growing attention in ovarian antiaging research, there are no clinically feasible strategies to either preserve or rejuvenate reproductive functions during aging ( 20 , 21 ).…”