IL-27 Mediates Pro-Inflammatory Effects via the ERK Signaling Pathway During Preterm Labor

Preterm labor (PTL) is a multifactorial syndrome that results in birth prior to 37 weeks of gestation. However, the specific molecular mechanisms underlying this condition have yet to be elucidated. Previous research demonstrated that the abnormal expression of IL-27, and its receptors, played a role in the pathophysiology of preterm labor. In the present study, we established a Lipopolysaccharide (LPS)-stimulated, infection-induced, preterm mouse model based on wild-type C57BL/6 mice and WSX-1-/-C57BL/6 mice. WSX-1 knockdown led to a significant delay in birth by 11.32 ± 2.157h. In addition, compared with wild-type C57B/6 mice, the expression levels of IFN-γ, IL-1β, IL-6, TNF-α, and CXCL10, in the fetal membrane and myometrium of WSX-1-/-mice were significantly lower, particularly in the myometrium. We also confirmed similar pro-inflammatory effects arising from IL-27 in human amniotic cell line (WISH) and human myometrial smooth muscle cell line (HMSMC). Once stimulated by LPS, the pro-inflammatory action exhibited a synergistic effect and appeared to be time-dependent. Finally, we demonstrated that LY3214996, an inhibitor of the ERK pathway, significantly inhibited the pro-inflammatory effect mediated by IL-27. Overall, our data confirmed that the inflammatory effect mediated by the IL-27/IFN-r/ERK axis is involved in preterm labor. Our findings, therefore, provide an enhancement in our etiological understanding of the mechanisms underlying PTL.

Current Trends in Orthobiologics: An 11-Year Review of the Orthopaedic Literature

Background: The use of “orthobiologics” or regenerative therapies in orthopaedic surgery has grown in recent years. Particular interest has been raised with regard to platelet-rich plasma, bone marrow aspirate, adipose-derived cells, and amniotic cells. Although studies have analyzed outcomes after orthobiologic treatment, no study has analyzed how the literature as a whole has evolved. Purpose: To evaluate trends in platelet-rich plasma, bone marrow aspirate, adipose-derived cells, and amniotic cell publications and to assess how these might inform efforts to establish minimum reporting standards and forecast future use. Study Design: Systematic review; Level of evidence, 4. Methods: A database was compiled systematically using PubMed to identify articles published between 2009 and 2019 within 9 prominent orthopaedic journals and pertaining to the use of platelet-rich plasma, bone marrow aspirate, adipose-derived cells, and amniotic cells in the treatment of musculoskeletal conditions. Included articles were classified as clinical, nonclinical (translational or basic science), or review, and a variety of study parameters were recorded for each. Additional queries were performed to identify articles that utilized minimum reporting standards. Results: A total of 474 articles (132 clinical, 271 nonclinical, 71 review) were included, consisting of 244 (51.5%) platelet-rich plasma, 146 (30.8%) bone marrow aspirate, 72 (15.2%) adipose-derived cells, and 12 (2.5%) amniotic cells. The greatest annual increase in publications for each orthobiologic topic was from 2018 to 2019. The American Journal of Sports Medicine demonstrated the highest number of overall (34.2%) and clinical (50.0%) publications, and accounted for 44.3% of all platelet-rich plasma publications. The Journal of Orthopaedic Research accounted for the second highest overall number of publications (24.9%) and highest nonclinical publications (41.0%). Platelet-rich plasma accounted for 91.5% of all level 1 clinical studies, while much greater than half of bone marrow aspirate, adipose-derived cells, and amniotic cell publications were level 3 or lower. Out of the 207 articles that used some form of reporting protocol, 59 (28.5%) used an established algorithm and 125 (60.4%) used their own. Conclusion: Interest in orthobiologics continues to grow, as evidenced by an increasing trend in publications over an 11-year period. However, current reporting on orthobiologic formulations is largely heterogeneous, emphasizing the need for minimum reporting standards and higher-quality studies.

The Immuno-Inflammatory Effect of IL-27 in Preterm Labor

Objective To investigate the effect of IL-27 on Th1 cells infiltration in human fetal membranes (FMs) and the underlying mechanisms in preterm labor. Methods The expression of IL-27 receptor a subunit (IL-27R𝛼) and Th1 cells were studied in human FMs from pregnant women with preterm labor (PL) and term labor (TL). In vitro, we investigated the effects of IL-27 in human amniotic cell lineage on the Th1 related chemokines and adhesive molecules and the underlying intracellular signaling molecules. Results The expression of IL-27R𝛼 was higher in human fetal membranes from PL group compared with TL group. Compared with TL group, the PL group had more Th1 cells infiltration in human FMs. Meanwhile, it was confirmed the expression of CXCL9, CXCL10, CXCL11 and ICAM-1 were higher in FMs from PL group to TL group. Moreover, IL-27 stimulation can up-regulate the expression of those chemokines in human amniotic cell lineage via JAK2/STAT1/STAT3 signaling pathway. Conclusions The expression of IL-27R𝛼 and number of Th1 cells infiltration were higher at FMs of PL group than TL group. IL-27 could promote Th1 related chemokines and adhesive molecules in human amniotic cell lineage mainly through JAK2/STAT1/STAT3 signaling pathway.

Secretome derived from different cell lines in bovine embryo production in vitro

The present study investigated the effects of conditioned medium (CM), composed of microvesicles (MVs) and soluble factors present in the supernatant (SN), of bovine endometrial and amniotic cells on embryo quality and rate of blastocyst production. Presumptive zygotes were randomly assigned on Days 1, 3 and 5 after fertilisation to synthetic oviducal fluid with amino acids (SOFaa; control) or to SOFaa supplemented with either 20% endometrial or amniotic CM, 20% SN or 100 × 106 MVs mL−1. Embryos were evaluated on Day 7. For groups supplemented with MVs derived from either endometrial or amniotic cells on Day 1 of culture, blastocysts had developed, but at a lower rate than in the control group. Blastocysts had developed in all groups in which endometrial or amniotic cell-derived CM or MVs were added on Day 3 of culture, but the rate of blastocyst development was significantly lower in both CM groups than in the MVs groups. The addition of all secretome fractions (CM, MVs and SN) derived from either bovine endometrial or amniotic cells on Day 5 of culture resulted in blastocyst production, but only amniotic MVs resulted in a blastocyst production rate comparable to that in the control group. Supplementation of SOFaa on Day 5 resulted in a qualitatively higher number of inner cell mass cells compared with the control group only for the amniotic CM and MVs groups. At day 7, these data were confirmed by RT-qPCR evaluation of genes (Bcl-2-associated X protein (BAX) and glutathione peroxidase 1 (GPX1) involved in apoptosis and protection against reactive oxygen species. In conclusion, of the different secretome fractions tested, only amniotic MVs added to SOFaa resulted in better outcomes than in the control group.

The Impact of Gestational Age on Targeted Amniotic Cell Profiling in Experimental Neural Tube Defects

Purpose: The proportions of select stem cells in term amniotic fluid have been shown to correlate with the type and size of experimental neural tube defects (NTDs). We sought to determine the impact of gestational age upon this form of targeted amniotic cell profiling. Methods: Sprague-Dawley fetuses with retinoic acid-induced NTDs (n = 110) underwent amniotic fluid procurement at four time points in gestation. Samples were analyzed by flow cytometry for the presence of cells concomitantly expressing Nestin and Sox-2 (neural stem cells, aNSCs) and cells concomitantly expressing CD29 and CD44 (mesenchymal stem cells, aMSCs). Statistical analysis was by nonparametric Kruskal-Wallis ANOVA (p < 0.05). Results: There was a statistically significant impact of gestational age on the proportions of both aMSCs (p = 0.01) and aNSCs (p < 0.01) in fetuses with isolated spina bifida. No such impact was noted in normal fetuses (p > 0.10 for both cells), in isolated exencephaly (p > 0.10 for both cells), or in combination defects (p > 0.10 for both cells). Gestational age had no effect on aNSC/aMSC ratios. Conclusions: Targeted quantitative amniotic cell profiling varies with gestational age in experimental isolated spina bifida. This finding should be considered prior to the eventual translation of this diagnostic adjunct into the prenatal evaluation of these anomalies.