Effects of Low-Intensity Pulsed Ultrasound on the Migration and Homing of Human Amnion–Derived Mesenchymal Stem Cells to Ovaries in Rats With Premature Ovarian Insufficiency

Abstract

Premature ovarian insufficiency (POI) can cause multiple sequelae and is currently incurable. Mesenchymal stem cell (MSC) transplantation might provide an effective treatment method for POI. However, the clinical application of systemic MSC transplantation is limited by the low efficiency of cell homing to target tissue in vivo, including systemic MSC transplantation for POI treatment. Thus, exploration of methods to promote MSC homing is necessary. This study was to investigate the effects of low-intensity pulsed ultrasound (LIPUS) on the migration and homing of transplanted human amnion–derived MSCs (hAD-MSCs) to ovaries in rats with chemotherapy-induced POI. For LIPUS treatment, hAD-MSCs were exposed to LIPUS or sham irradiation. Chemokine receptor expressions in hAD-MSCs were detected by polymerase chain reaction (PCR), Western blot, and immunofluorescence assays. hAD-MSC migration was detected by wound healing and transwell migration assays. Cyclophosphamide-induced POI rat models were established to evaluate the effects of LIPUS on the homing of systemically transplanted hAD-MSCs to chemotherapy-induced POI ovaries in vivo. We found that hAD-MSCs expressed chemokine receptors. The LIPUS promoted the expression of chemokine receptors, especially CXCR4, in hAD-MSCs. SDF-1 induced hAD-MSC migration. The LIPUS promoted hAD-MSC migration induced by SDF-1 through SDF-1/CXCR4 axis. SDF-1 levels significantly increased in ovaries induced by chemotherapy in POI rats. Pretreating hAD-MSCs with LIPUS increased the number of hAD-MSCs homing to ovaries in rats with chemotherapy-induced POI to some extent. However, the difference was not significant. Both hAD-MSC and LIPUS-pretreated hAD-MSC transplantation reduced ovarian injuries and improved ovarian function in rats with chemotherapy-induced POI. CXCR4 antagonist significantly reduced the number of hAD-MSCs- and LIPUS-pretreated hAD-MSCs homing to POI ovaries, and further reduced their efficacy in POI treatment. According to these findings, pretreating MSCs with LIPUS before transplantation might provide a novel, convenient, and safe technique to explore for improving the homing of systemically transplanted MSCs to target tissue.

Effects of Ginsenoside Rg1 on the Biological Behavior of Human Amnion-Derived Mesenchymal Stem/Stromal Cells (hAD-MSCs)

“…Primary hAD-MSCs were isolated, cultured, and identified according to our previous published research [ 4 , 17 ]. Donors ( n = 18) were healthy full-term pregnant women from the Second Affiliated Hospital of Chongqing Medical University, China.…”

“…Image-Pro Plus 6.0 software (Media Cybernetics, Bethesda, MD, USA) was used to count Hoechst33342 and EdU positive cells. hAD-MSC proliferation rate was defined as EdU positive cells (red cells)/Hoechst33342 positive cells (blue cells) [ 4 ].…”

“…MSCs can be isolated from a broad subset of tissue sources, such as amniotic membrane, amniotic fluid, umbilical cord, bone marrow, dental pulp, and adipose tissue [ 1 , 2 ]. In previous studies, MSCs have been explored to treat various diseases [ 3 , 4 ]. Recently, MSC transplantation has become a promising and focal treatment for the regeneration of injured tissue and refractory diseases [ 1 , 4 , 5 ].…”

“…In previous studies, MSCs have been explored to treat various diseases [ 3 , 4 ]. Recently, MSC transplantation has become a promising and focal treatment for the regeneration of injured tissue and refractory diseases [ 1 , 4 , 5 ].…”

“…During the expansion of MSCs in vitro , there are some uncontrolled factors, such as the limited number of primary MSCs [ 2 , 10 ], the decreases of proliferation and differentiation capacity of MSCs after passaging, and the interdonor variability and scalability associated with primary donor-derived MSCs [ 4 , 11 , 12 ], which may affect the number and quality of MSCs before clinical utility [ 1 , 2 , 13 ]. During the transplantation of MSCs, there are still various unsolved problems, such as the loss of MSCs, low homing and engraftment rates of MSCs, inability of precise regulation of MSC differentiation, and accelerated senescence and apoptosis of MSCs after cell transplantation, which are major issues in MSC preclinical research and may influence the clinical utility of MSCs [ 1 – 3 ].…”