Human umbilical cord platelet-rich plasma to treat endometrial pathologies: methodology, composition and pre-clinical models

Abstract

STUDY QUESTION Can human umbilical cord platelet-rich plasma (hUC-PRP) efficiently treat endometrial damage and restore fertility in a preclinical murine model? SUMMARY ANSWER Local application of hUC-PRP promotes tissue regeneration and fertility restoration in a murine model of Asherman syndrome and endometrial atrophy (AS/EA). WHAT IS KNOWN ALREADY AS/EA are well-described endometrial pathologies that cause infertility, however, there are currently no gold standard treatments available. Recent reports have described the successful use of human platelet-rich plasma (hPRP) in reproductive medicine, and its regenerative potential is further enhanced using hUC-PRP, due to the ample growth factors and reduced pro-inflammatory cytokines in the latter. STUDY DESIGN, SIZE, DURATION hUC-PRP (n = 3) was processed, characterized, and delivered locally to endometrial damage in a murine model (n = 50). The hUC-PRP was either used alone or loaded into a decellularized porcine endometrium-derived extracellular matrix (EndoECM) hydrogel; endometrial regeneration, fertility outcomes and immunocompatibility were evaluated two weeks following treatment administration. PARTICIPANTS/MATERIALS, SETTING, METHODS Umbilical cord blood was obtained from women in childbirth. Endometrial damage (mimicking AS/EA) was induced using ethanol in eight-week-old C57BL/6 mice, and treated with the most concentrated hUC-PRP sample four days later. Characterization of hUC-PRP and immunotolerance was carried out with multiplex technology, while uterine samples were analyzed by immunohistochemistry and quantitative PCR. The number of embryos and their morphology was determined visually. LARGE SCALE DATA N/A. MAIN RESULTS AND THE ROLE OF CHANCE Platelet density was enhanced three-fold in hUC-PRP compared to that in hUC blood (P < 0.05). hUC-PRP was enriched with growth factors related to tissue regeneration (i.e., hepatocyte growth factor (HGF), platelet-derived growth factor (PDGF-BB), and epidermal growth factor (EGF)), which were released constantly (in vitro) when hUC-PRP was loaded into EndoECM. Both treatments (hUC-PRP alone and hUC-PRP with EndoECM) were immunotolerated and caused significantly regeneration of the damaged endometrium, evidenced by increased endometrial area, neoangiogenesis, cell proliferation and gland density and lower collagen deposition with respect to non-treated uterine horns (P < 0.05). Additionally, we detected augmented gene expression of Akt1, VEGF, and Ang, which are involved in regenerative and proliferation pathways. Finally, hUC-PRP treatment restored pregnancy rates in the mouse model. LIMITATIONS, REASONS FOR CAUTION This proof-of-concept pilot study was based on a murine model of endometrial damage and the use of EndoECM requires further validation prior to clinical implementation for women affected by AS/EA. WIDER IMPLICATIONS OF THE FINDINGS The local administration of hUC-PRP has high impact and is immunotolerated in a murine model of AS/EA, as has been reported in other tissues, making it a promising candidate for heterologous treatment of these endometrial pathologies. STUDY FUNDING/COMPETING INTEREST(S) This study was supported by the Ministerio de Ciencia, Innovación y Universidades; Conselleria de Innovación, Universidades, Ciencia y Sociedad Digital, Generalitat Valenciana; and Instituto de Salud Carlos III. The authors do not have any conflicts of interest to declare.

Plasma‐derived from human umbilical cord blood restores ovarian function and improves serum reproductive hormones levels in mice with premature ovarian insufficiency (POI) through cytokines and growth factors

“…Recent study have found that CBP contains low concentrations of proinflammatory cytokines (IL‐2, IL‐6, IFN‐γ, and TNF‐α), elevated chemokine IL‐8 and higher levels of VEGF, G‐CSF, EGF, and FGF‐basic growth factors, which promote mononuclear cell viability and reduced apoptotic cell death in vitro and improve neurodegenerative diseases (Ehrhart et al, 2018). Further, several studies have explored the effects of cytokines on ovarian function (Rodríguez‐Eguren et al, 2023; Said et al, 2016). Recent studies have shown that the messenger RNA (mRNA) levels of IL‐6 and IL‐8 in ovarian tissue are significantly upregulated, while concomitant reduction in ovarian anti‐inflammatory cytokine IL‐10 mRNA level (Said et al, 2016; Walsh et al, 2013).…”

“…The immune regulatory network can play an essential role in the restoration for POI, which achieves the treatment by modulating the inflammatory microenvironment (Huang et al, 2021). Previous studies have shown that cytokines and growth factors in PRP derived from hUCB can effectively improve ovarian function and hormone levels in POI (Cakiroglu et al, 2020; Panda et al, 2020; Rodríguez‐Eguren et al, 2023; Sfakianoudis et al, 2020). It is clear from previous research that CBP was shown to be richer for cytokines and growth factors compared to ABP and PRP (Ehrhart et al, 2018; Rhéaume et al, 2022).…”

Recent advances in platelet-rich plasma and its derivatives: therapeutic agents for tissue engineering and regenerative medicine

“…In a pilot investigation, C57BL/6 mice bearing the Asherman syndrome and endometrial atrophy were utilized to evaluate the effectiveness of human umbilical cord PRP in tissue regeneration and reproductive restoration. In the murine model, it was found that human Umbilical Cord PRP (hUC-PRP) increased reproductive rates when infused into the uterine cavity on the thirteenth day of the HRT cycle [171].…”

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

“…Their proteomic analyses revealed overexpression of HOXA10, related to endometrial functionality, and proteins involved in angiogenesis, mitotic cell cycle, PI3K/AKT and JAK/STAT signaling pathways. Rodríguez-Eguren et al [231] characterized thoroughly the composition of hUC-PRP processed after manual and commercial strategies. Then, its regenerative effect alone or driven by the aforementioned EndoECM was evaluated in a mouse model of AS/EA.…”