Vitamin C plus hydrogel facilitates bone marrow stromal cell-mediated endometrium regeneration in rats

Abstract

BackgroundIntrauterine adhesion (IUA) is a common uterine cavity disease which can be caused by mechanical damage that may eventually lead to infertility and pregnancy abnormalities. Since the effect of therapeutic drugs appears disappointing, cell therapy has emerged as an alternative choice for endometrium regeneration. The aim of this study is to investigate whether the combination of hydrogel Pluronic F-127 (PF-127), Vitamin C (Vc), and a bone marrow stromal cell (BMSC) mixture could be a feasible strategy to improve the endometrial regeneration in a mechanical damage model of IUA in rats.MethodsFirstly, PF-127 cytotoxicity and the effect of Vc was tested in vitro using the Annexin V/propidium iodide (PI) apoptosis test, cell count kit (CCK) growth test, and enzyme-linked immunosorbent assay (ELISA). For the establishment of the rat IUA model, a 2-mm transverse incision in the uterus was prepared at the upper end, and 1.5- to 2.0-cm endometrial damage was scraped. Rats were randomly assigned to five groups to investigate the combined strategy on IUA uterine regeneration: a sham group, an IUA control group, an IUA BMSC encapsulated in PF-127 plus Vc group, an IUA BMSC plus Vc group, and an IUA PF-127 plus Vc group. A cell mixture was injected into the uterine horn while making the IUA model. Eight weeks after cell transplantation, the rats were sacrificed and the uterine was dissected for analysis. Endometrial thickness, gland number, fibrosis area, and the expression of marker proteins for endometrial membrane were examined by hematoxylin and eosin staining, Masson’s staining, and immunohistochemistry.ResultsVc promoted the survival and health of PF-127-encapsulated BMSCs in vitro. When this combination was transplanted in vivo, the endometrium showed better restoration as the endometrium membrane became thicker and had more glands and less fibrosis areas. The expression of cytokeratin, von Willebrand Factor (vWF), was also restored. The proinflammatory cytokine interleukin-1β (IL-1β) was significantly lower compared with the control group.ConclusionsVc alleviates the cytotoxic effect of PF-127 and promotes cell survival and growth in rat BMSC encapsulation. Thus, a cell therapy strategy containing biomaterial scaffold, BMSCs and the modulatory factor Vc promotes the restoration of damaged IUA endometrium.

PF-127 hydrogel plus sodium ascorbyl phosphate improves Wharton’s jelly mesenchymal stem cell-mediated skin wound healing in mice

“…It has been reported that several kinds of cell membrane-stabilizing agents such as hydrocortisone and vitamin C are added to the PF-127 gel formulations to increase cell viability [24,32]. In this study, we proposed vitamin C analog SAP as a new effective cell membranestabilizing agent.…”

“…Several studies have shown the therapeutic benefits of PF-127 as cell encapsulation scaffold in tissue engineering, and it has been used as scaffolds for porcine chondrocytes, somatic lung progenitor cells, and human adipose-derived stem cells [56][57][58]. However, it has also been reported that PF-127 encapsulation exerts a negative effect on cell viability, which is potentially destructive to cell-based therapy [23,32]. Indeed, we demonstrated a 15.80 ± 2.59% decrease in cell survival at a concentration of 20% (w/v) PF-127 encapsulation after 24 h. Further, in vivo animal experiments displayed no statistically significant improvement on wound healing in the WJMSCs/PF-127 group as compared to the WJMSCs group.…”

“…PF-127 hydrogel preparation was performed according to previous report [32]. Briefly, 20% (w/v) PF-127 solution was dissolved in DMEM-F12 medium at 4°C, then filtered through a 0.22 μm filter (Millipore, USA), and finally kept at 4°C for use.…”

Advances in the Application of Biomimetic Endometrium Interfaces for Uterine Bioengineering in Female Infertility

“…BMSC-loaded scaffold recovered the receptive ability of the new endometrium. Yang et al (2017) reported that BMSCs were encapsulated by thermo-responsive gelation of pluronic F-127 (PF-127) and vitamin C, which added to the membrane stability. In addition, vitamin C, as a prominent antioxidant, downregulated tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6) secretion, maintained redox homeostasis and facilitated a pro-regenerative tendency by increasing interleukin 10 (IL-10) level (El Banna et al, 2019;Qi et al, 2019).…”

Cell‐Surface Engineering for Advanced Cell Therapy

“…In addition to alginate‐based ionotropic hydrogels and covalently cross‐linked hydrogels, other chemical interactions have been introduced to encapsulate cells. These include supramolecular host‐guest interactions (host: cyclodextrins or cucurbiturils; guest: adamantanes or diaminohexanes), London‐dispersion forces (hydrophobic effect‐driven self‐assembly), catecholic chemistry, and primarily hydrogen bonding using agarose or pluronic F‐127 . Table summarized cell encapsulation in a covalently cross‐linked hydrogel.…”