Newborn Tissue Research Articles

Metabolomic signatures associated with fetal growth restriction and small for gestational age: a systematic review.

The pathways involved in the pathophysiology of fetal growth restriction (FGR) and small for gestational age (SGA) are incompletely understood. We conduct a systematic review to identify metabolomic signatures in maternal and newborn tissues and body fluids samples associated with FGR/SGA. Here, we report that 825 non-duplicated metabolites were significantly altered across the 48 included studies using 10 different human biological samples, of which only 56 (17 amino acids, 12 acylcarnitines, 11 glycerophosphocholines, six fatty acids, two hydroxy acids, and eight other metabolites) were significantly and consistently up- or down-regulated in more than one study. Three amino acid metabolism-related pathways and one related with lipid metabolism are significantly associated with FGR and/or SGA: biosynthesis of unsaturated fatty acids in umbilical cord blood, and phenylalanine, tyrosine and tryptophan biosynthesis, valine, leucine and isoleucine biosynthesis, and phenylalanine metabolism in newborn dried blood spot. Significantly enriched metabolic pathways were not identified in the remaining biological samples. Whether these metabolites are in the causal pathways or are biomarkers of fetal nutritional deficiency needs to be explored in large, well-phenotyped cohorts.

Pre-pregnancy BMI, gestational weight gain, and telomere length in amniotic fluid: a causal graph analysis.

Previous investigations have suggested a potential association between pre-pregnancy body mass index (BMI) and gestational weight gain (GWG) with telomere length (TL) in various tissues of pregnant women and newborns. Nonetheless, as association does not imply causation, our objective was to investigate the causal connections among pre-pregnancy BMI, GWG, and TL in amniotic fluid. The analysis included 136 mother-child pairs from the Mamma & Bambino cohort, and three causal graph models were developed to depict the interconnections between pre-pregnancy BMI, GWG, and TL. Causal graph analysis was conducted utilizing the do-operator to estimate the causal effect of GWG and the controlled direct effect of pregestational BMI. We revealed that transitioning from non-adequate to adequate GWG had a positive impact on the probability of having "long" TL (i.e., a value greater than the population median) in all three models. When considering the effect of pre-pregnancy BMI, the highest probability of "long" TL was observed in normal weight women with adequate GWG. In contrast, the effect of adequate GWG became minimal among overweight women. These results shed light on the potential causality between pre-pregnancy BMI, GWG, and TL in amniotic fluid, emphasizing the importance of appropriate weight management before and during pregnancy for optimal TL outcomes.

Micro–nano hierarchical scaffold providing temporal-matched biological constraints for tendon reconstruction

Due to the limitations of tendon biology, high-quality tendon repair remains a clinical and scientific challenge. Here, a micro–nano hierarchical scaffold is developed to promote orderly tendon regeneration by providing temporal-matched biological constraints. In short, fibrin (Fb), which provides biological constraints, is loaded into poly (DL-lactide-co-glycolide) nanoyarns with suitable degradation cycles (Fb-loaded nanofiber yarns (Fb-NY)). Then further combined with braiding technology, temporary chemotactic Fb scaffolds with tendon extracellular matrix-like structures are obtained to initiate the regeneration process. At the early stage of healing (2 w), the regeneration microenvironment is regulated (inducing M2 macrophages and restoring the early blood supply necessary for healing) by Fb, and the alignment of cells and collagen is induced by nanoyarn. At the late healing stage (8 w), with the degradation of Fb-NY, non-functional vascular regression occurs, and the newborn tissues gradually undergo load-bearing remodeling, restoring the anvascularous and ordered structure of the tendon. In summary, the proposed repair strategy provides temporal-matched biological constraints, offering a potential pathway to reconstruct the ordered structure and function of tendons.

Evaluating the Pharmacokinetics of Fentanyl in the Brain Extracellular Fluid, Saliva, Urine, and Plasma of Newborns from Transplacental Exposure from Parturient Mothers Dosed with Epidural Fentanyl Utilizing PBPK Modeling.

Fentanyl can mitigate the mother and newborn complications resulting from labor pain. However, fentanyl shows a narrow therapeutic index between its respiratory depressive and analgesic effects. Thus, prenatally acquired high fentanyl levels in the newborn brain extracellular fluid (bECF) may induce respiratory depression which requires therapeutic drug monitoring (TDM). TDM using saliva and urine in newborns can reduce the possibility of infections and distress associated with TDM using blood. The objective of this study was to develop aphysiologically based pharmacokinetic (PBPK) modelto predict fentanyl concentrations in different newborn tissues due to intrauterine exposure. A fentanyl PBPK model in adults after intravenous and epidural administration was built, validated, and scaled to pregnancy and newborn populations. The dose that the newborn received transplacentally at birth was calculated using the pregnancy model. Then, the newborn bECF, saliva, plasma, and urine concentrations after such a dose were predicted using the newborn PBPK model. After a maternal epidural dose of fentanyl245 µg, the predicted newborn plasma and bECF levels were below the toxicity thresholds. Furthermore, the salivary threshold levels in newborns for fentanyl analgesic and respiratory depression effects were estimated to be 0.39 and 14.7-18.2 ng/ml, respectively. The salivary TDM of fentanyl in newborns can be useful in newborns exposed to intrauterine exposure from parturient females dosed with epidural fentanyl. However, newborn-specific values of µ-opioid receptors IC50 for respiratory depression are needed.

30-LB: Study on the Mechanism of Calycosin in Promoting Wound Healing in Diabetic Foot Ulcers

Objective: To investigate the effect of calycosin in promoting wound healing of foot ulcers and explore the regulation on vascular endothelial growth factor (VEGF). Methods: 8-week-old male C57BL/6J mice were injected intraperitoneally with 1% streptozotocin 120 mg/kg for 3 consecutive times, and the mice with random blood glucose > 16.7 mmol/L were used and randomly divided into model group and calycosin suspension groups (30 mg/kg, 60 mg/kg, 90 mg/kg). Mice were pretreated for 30 days at a dose of 10 mL/kg of the corresponding drugs once a day, and the control group was given the same dose of 0.5% sodium carboxymethyl cellulose solution. At the end of administration, the animals were anesthetized with isoflurane and the whole layer of skin with diameter 10 mm was removed on the back to establish the wound model of diabetic foot ulcer as day 0. The wound healing was observed and photographed with a camera on the day 0, 2, 4, 6, 8, 10 and 12, respectively. At the end of the day 12, the new tissues around the wound were taken for the detections of the growth of new granulation tissue, VEGF, CD31 and the content of sorbitol. Results: Compared with the model group, the wound healed faster in calycosin groups. It closed on the 12th day, but the wound in the model group did not heal completely. However, there was no obvious dose-effect relationship between different dose groups. The mice in treatment groups showed a better structural integrity of epidermis, and a more expression of VEGF and CD31 in the newborn tissue. And the protein level of VEGF of the calycosin (60 mg/kg) group increased. And the sorbitol content in calycosin (90 mg/kg) group reduced significantly (P<0.05), while in 30 mg/kg and 60 mg/kg changed slightly. Conclusion: Calycosin could promote the wound healing of diabetic foot ulcer, which may be related to the effect of increasing the level of VEGF and reducing the accumulation of sorbitol in tissues. And further research was needed for fully exploration on the mechanism. Disclosure Y. Xu: None. J. Ke: None. L. Yang: None. D. Zhao: None. Funding Tongzhou District Science and Technology Plan Project (KJ2022CX024)

Assessing maternal and fetal SARS-CoV-2 viral load, antibody profiles and placental pathology following prenatal infection

Evaluate mother-to-infant transmission potential of SARS-CoV-2 and patterns of antibody transfer following prenatal infection. Prospective cohort of pregnant individuals infected with COVID-19. Samples collected at delivery included maternal swabs (nasopharyngeal/oropharyngeal, vaginal, anorectal), blood, breastmilk, newborn swabs (nasopharyngeal, sub-amniotic), amniotic fluid, cord blood, and placenta. Samples were analyzed by qPCR for SARS-CoV-2-RNA and IgG assays targeting the SARS-CoV-2 spike (S), nucleocapsid (N), and receptor binding domain (RBD) proteins. Placentas were evaluated for histologic lesions. The cohort included 213 individuals who delivered between from March 2020 to August 2021, and their 215 exposed offspring. SARS-CoV-2 was evident in maternal nasopharyngeal (64/145), vaginal (3/184) and anorectal (5/182) swabs and breastmilk (1/38). A total of 27/212 infants (12.7%) demonstrated evidence of mother-to-child transmission of SARS-CoV-2; SARS-CoV-2 was detected in nasopharyngeal swabs (21/142), amniotic fluid (3/77), sub-amniotic swabs (1/165), and placenta homogenates (6/138). Anti-SARS-CoV-2 IgA and IgM were detected in 1/189 neonates. Maternal and cord serum anti-RBD, anti-N and anti-S IgG were significantly and positively correlated ( R =0.26, 0.24, 0.23, respectively; P <0.05). Antibody transfer ratio of anti-RBD IgG was significantly correlated with weeks since maternal infection during pregnancy (R = 0.28, P <0.05). SARS-CoV-2 was detected in 6/138 placentas; placentas were characterized by acute (33/113), chronic inflammation (20/113), maternal (39/113) and fetal (12/113) vascular malperfusion. The rate of SARS-CoV-2 vertical transmission and transfer of maternal antibodies to newborn tissues was high. There were no pathological differences between infected placentas and controls. Further investigation into the timing of vertical transmission is warranted and its potential implications on offspring outcomes.

Maturation state of colonization sites promotes symbiotic resiliency in the Euprymna scolopes-Vibrio fischeri partnership

BackgroundMany animals and plants acquire their coevolved symbiotic partners shortly post-embryonic development. Thus, during embryogenesis, cellular features must be developed that will promote both symbiont colonization of the appropriate tissues, as well as persistence at those sites. While variation in the degree of maturation occurs in newborn tissues, little is unknown about how this variation influences the establishment and persistence of host-microbe associations.ResultsThe binary symbiosis model, the squid-vibrio (Euprymna scolopes-Vibrio fischeri) system, offers a way to study how an environmental gram-negative bacterium establishes a beneficial, persistent, extracellular colonization of an animal host. Here, we show that bacterial symbionts occupy six different colonization sites in the light-emitting organ of the host that have both distinct morphologies and responses to antibiotic treatment. Vibrio fischeri was most resilient to antibiotic disturbance when contained within the smallest and least mature colonization sites. We show that this variability in crypt development at the time of hatching allows the immature sites to act as a symbiont reservoir that has the potential to reseed the more mature sites in the host organ when they have been cleared by antibiotic treatment. This strategy may produce an ecologically significant resiliency to the association.ConclusionsThe data presented here provide evidence that the evolution of the squid-vibrio association has been selected for a nascent organ with a range of host tissue maturity at the onset of symbiosis. The resulting variation in physical and chemical environments results in a spectrum of host-symbiont interactions, notably, variation in susceptibility to environmental disturbance. This “insurance policy” provides resiliency to the symbiosis during the critical period of its early development. While differences in tissue maturity at birth have been documented in other animals, such as along the infant gut tract of mammals, the impact of this variation on host-microbiome interactions has not been studied. Because a wide variety of symbiosis characters are highly conserved over animal evolution, studies of the squid-vibrio association have the promise of providing insights into basic strategies that ensure successful bacterial passage between hosts in horizontally transmitted symbioses.2h6LgmG9jj4pmTHJMXeiS6Video

In situ fabrication of an anisotropic double-layer hydrogel as a bio-scaffold for repairing articular cartilage and subchondral bone injuries.

Articular cartilage is a smooth and elastic connective tissue playing load-bearing and lubricating roles in the human body. Normal articular cartilage comprises no blood vessels, lymphatic vessels, nerves, or undifferentiated cells, so damage self-repair is very unlikely. The injuries of articular cartilage are often accompanied by damage to the subchondral bone. The subchondral bone mainly provides mechanical support for the joint, and the successful repair of articular cartilage depends on the ability of the subchondral bone to provide a suitable environment. Currently, conventional repair treatments for articular cartilage and subchondral bone defects can hardly achieve good results due to the poor self-repairing ability of the cartilage Here, we propose a bioactive injectable double-layer hydrogel to repair articular cartilage and subchondral bone. The hydrogel scaffold mimics the multilayer structure of articular cartilage and subchondral bone. Agarose was used as a common base material for the double-layer hydrogel scaffold, in which a sodium alginate (SA)/agarose layer was used for the repair of artificially produced subchondral bone defects, while a decellularized extracellular matrix (dECM)/agarose layer was used for the repair of articular cartilage defects. The double-layer hydrogel scaffold is injectable, easy to use, and can fill in the damaged area. The hydrogel scaffold is also anisotropic both chemically and structurally. Animal experiments showed that the surface of the new cartilage tissue in the double-layer hydrogel scaffold group was closest to normal articular cartilage, with a structure similar to that of hyaline cartilage and a preliminary calcified layer. Moreover, the new subchondral bone in this group exhibited many regular bone trabeculae, and the new cartilage and subchondral bone were mechanically bound without mutual intrusion and tightly integrated with the surrounding tissue. The continuous double-layer hydrogel scaffold prepared in this study mimics the multilayer structure of articular cartilage and subchondral bone and promotes the functional repair of articular cartilage and subchondral bone, favoring close integration between the newborn tissue and the original tissue.

Comparison of Healing Effect Between Fibrin Gel Complex Bone Morphological Protein and Reconstituted Bone Xenograft After Reconstruction of Sports Ligament Injury

To study the effect of fibrin gel complex bone morphological protein (FG-BMP) and RBX on the healing of tendon-bone interface. 51 live rabbits were selected, and 51 rabbits were randomly divided into 3 groups: FG-BMP, RBX and control groups. Bilateral ACL reconstruction was performed in 51 rabbits, with different biomaterials injected at the tendon-bone interface, and none applied in the FG-BMP, RBX, and control groups. Material extraction, imaging examination, histological analysis, and biomechanical tests were conducted at 2 weeks after surgery, 6 weeks later, and 12 weeks after surgery, respectively. The tendon-interbone growth rate of rabbits using FG-BMP biomaterials and RBX biomaterials was significantly improved in the FG-BMP and RBX groups compared with the control group (P &lt; 0.05). Histological results showed that all rabbits in the RBX group were surrounded by newborn bone tissue around the tendon-bone transplantation tendon and had a significantly greater number of newborn bone tissue than in the FG-BMP group (P &lt; 0.05). The biomechanical test results showed that the two groups of FG-BMP biomaterials improved the maximum tension compared with the control group (P &lt; 0.05), where the RBX tension, the difference was significant (P &lt; 0.05), and the FG-BMP group although higher than the control group (P &gt; 0.05). Conclusion: In the comparison of promoting tendon-bone interface healing, FG-BMP biomaterials and RBX biomaterials have significant results, especially RBX materials have obvious advantages in accelerating tendon-bone interface healing.

Maternal vaccination with a type-III glycoconjugate protects mouse neonates against Group B Streptococcus intranasal infection

Group B Streptococcus (GBS) is generally an asymptomatic colonizer of human mucosa but it occasionally infects pregnant women and neonates through vertical transmission, causing disease during the first weeks of life with frequent and severe complications. Preclinical studies have shown that maternal vaccination with polysaccharide-based vaccines protects mothers and offspring from GBS mucosal colonization and consecutive infection. In these models, bacteria were inoculated in mouse either intravaginally in the last trimester of pregnancy or systemically in pups. Here, we investigated whether maternal vaccination with glycoconjugate vaccines may also prevent GBS-mediated colonization and disease in neonates using an infection route that more closely mimics inhalation or ingestion of bacteria during human delivery. To address this point, mice aged less than two days were intranasally challenged with epidemiologically relevant GBS strains. Bacteria were found to colonize nose and intestine, reaching in some cases lungs and blood during the first days of life. Bacteria were also found in vagina of a fraction of colonized female mice within the first month of life. GBS-specific IgG induced by maternal vaccination with a glycoconjugate vaccine formulation were found in blood and mucosal tissues of newborns. Finally, when intranasally challenged with GBS serotype III strains, pups delivered by vaccinated mothers were partially protected against mucosal colonization and deeper infection.

Comprehensive Analysis of miRNAs and Target mRNAs between Immature and Mature Testis Tissue in Chinese Red Steppes Cattle.

Simple SummaryMicroRNAs are small molecules that can regulate the relative abundance of their target genes by binding to the 3′ untranslated region of the target genes at the post-transcriptional level to affect various biological processes, such as biosynthesis, fat metabolism and proliferation, apoptosis, and cell differentiation. Fertility is one of the most important economic traits in livestock production. Bulls require the continuous production of high-quality spermatozoa in abundance. The quality of semen is an exceptionally important factor affecting the fertilization rate of the dairy cow and is also associated with the increasing conception rate in the process of artificial insemination. Therefore, accurately predicting fertility potential for a semen sample from donor bull for artificial insemination is crucial for consistently high reproductive efficiency. The present study performed a genome-wide sequencing analysis of microRNAs and mRNAs between immature and mature testes of Chinese Red Steppes. These results provide novel candidate microRNAs and functional genes related to bull reproduction traits and the networks between microRNAs and target genes, which will provide a useful genetic mechanism and epigenetic information for marker-assisted selection of bulls with excellent sperm quality in the future.This study aims to screen potential regulators and regulate fecundity networks between microRNAs (miRNAs) and target genes. The bovine testes of immature and mature Chinese Red Steppes were performed by genome-wide analysis of mRNAs and miRNAs. Compared with testicular tissues of newborns, 6051 upregulated genes and 7104 downregulated genes in adult cattle were identified as differentially expressed genes (DEGs). The DEGs were significantly enriched in 808 GO terms (p < 0.05) including male gonad development, male genitalia development, spermatogenesis, and sperm motility. Moreover, DEGs were also significantly enriched in 105 KEGG pathways (p < 0.05), including cGMP-PKG signaling pathway and calcium signaling pathway. To explore the expression of miRNA-regulated gene expression, 896 differentially expressed target genes negatively regulated with the expression levels of 31 differentially expressed miRNAs (DERs) were predicted and analyzed, and a network-integrated analysis was constructed. Furthermore, real-time PCR was performed to verify the expression levels of DEGs and DERs. Our results identified novel candidate DEGs and DERs correlated with male reproduction and intricate regulating networks between miRNAs and genes, which will be valuable for future genetic and epigenetic studies of sperm development and maturity, as well as providing valuable insights into the molecular mechanisms of male fertility and spermatogenesis in cattle.

A study of dosimetric parameters of new born tissue and adult tissue of some organs

We have calculated the radiological parameters such as computerized tomography (CT) numbers effective atomic number (Zeff) and electron density (Nel) of newborn tissue and adult tissue of lung, kidney, heart and brain. Effective atomic number is computed by calculating the molecular, atomic and electronic cross section in the wide energy range of 1 keV–100 keV using WinXCOM. The new born tissue and adult tissue a very similar, but their atomic number, CT number and electron densities differs. This fact helps in the radiography and radiotherapy.

Telomerase activation in the treatment of aging or degenerative diseases: a systematic review.

Telomeres are protective structures that are shortened during the lifetime, resulting in aging and degenerative diseases. Subjects experiencing aging and degenerative disorders present smaller telomeres than young and healthy ones. The size of these structures can be stabilized by telomerase, an enzyme which is inactive in adult tissues but functional in fetal and newborn tissues and adult testes and ovaries. The aim of this study was to perform a systematic review to evaluate the effect of telomerase activation in the treatment of degenerative and aging disorders. We accomplished the search using the Pubmed interface for papers published from September 1985 to April 16th, 2020. We found twenty one studies that matched our eligibility criteria. I concluded that telomerase is probably a potential and safe treatment for aging and degenerative diseases, demonstrating neither side effects nor risk of cancer in the selected studies. Further studies in humans are needed to confirm safety and efficiency of this treatment.

Hypoxia inutero increases the risk of pulmonary hypertension in rat offspring and is associated with vasopressin type‑2 receptor upregulation.

Pulmonary hypertension (PH) in newborns and adults is a disease that can lead to right heart failure and result in a shorter lifespan. PH was induced by maintaining pregnant rats in a hypoxic chamber for 4 h twice a day, from days 7–21 of pregnancy. Hypoxia was confirmed by a decrease in the partial pressure of oxygen (PaO2) and the oxygen saturation (SaO2) of arterial blood in the aorta. The body weight of newborns from hypoxic rats was ~20% decreased compared with the control newborns of normoxic rats. The vascular wall thickness/vascular diameter values of hypoxia treated pubs were increased compared with that of control newborns 7 days after birth; however, it decreased to similar levels than in the control group after 3 months, and then further decreased to significantly lower levels than in the control group at 6 months after birth. At birth, the lung tissues of newborns from hypoxic rats exhibited an increase in the levels of mRNA and proteins associated with PH such as HIF-1α, HIF-2α, V2R, TGF-β, TNF-α, Ang-2 and α-SMA. At 3 and 6 months after birth, the levels of both V2R mRNA and protein in offspring from hypoxic rats were at least 2-fold higher, whereas the expression of all other factors decreased compared with the control offspring. By contrast, HIF-2α and Ang-2 expression levels were significantly increased in the 6-month-old control offspring from normoxic rats. V2R overexpression in pups induced by hypoxia in maternal rats was sustained until their adulthood. V2R may be a marker for detecting PH.

The tissue-specific aspect of genome-wide DNA methylation in newborn and placental tissues: implications for epigenetic epidemiologic studies.

Epigenetic programming is essential for lineage differentiation, embryogenesis and placentation in early pregnancy. In epigenetic association studies, DNA methylation is often examined in DNA derived from white blood cells, although its validity to other tissues of interest remains questionable. Therefore, we investigated the tissue specificity of epigenome-wide DNA methylation in newborn and placental tissues. Umbilical cord white blood cells (UC-WBC, n = 25), umbilical cord blood mononuclear cells (UC-MNC, n = 10), human umbilical vein endothelial cells (HUVEC, n = 25) and placental tissue (n = 25) were obtained from 36 uncomplicated pregnancies. Genome-wide DNA methylation was measured by the Illumina HumanMethylation450K BeadChip. Using UC-WBC as a reference tissue, we identified 3595 HUVEC tissue-specific differentially methylated regions (tDMRs) and 11,938 placental tDMRs. Functional enrichment analysis showed that HUVEC and placental tDMRs were involved in embryogenesis, vascular development and regulation of gene expression. No tDMRs were identified in UC-MNC. In conclusion, the extensive amount of genome-wide HUVEC and placental tDMRs underlines the relevance of tissue-specific approaches in future epigenetic association studies, or the use of validated representative tissues for a certain disease of interest, if available. To this purpose, we herewith provide a relevant dataset of paired, tissue-specific, genome-wide methylation measurements in newborn tissues.

Age‐ and Tissue‐Specific Alternative Splicing of the Neurofibromin Gene in Swine

Neurofibromatosis type 1 (NF1) is one of the most common monogenic disorders that affects 1 in 2,500 children. The disorder manifests in varying, complex phenotypes that include cognitive and learning dysfunction, bone and cardiovascular disorders, and an increased risk of developing benign and malignant nervous system tumors. NF1 is caused by mutations in the neurofibromin gene; over 4,000 different mutations have been clinically identified with very little understanding of genotype‐phenotype association in terms of the clinical presentation of the disorder. The human neurofibromin gene contains 61 exons, of which, four are alternatively spliced (optionally included) to produce isoforms of the protein. Over 50% of the mutations in NF1 patients are thought to produce abnormalities in splicing. Yet, only a handful of studies have examined the role of neurofibromin alternative splicing or acknowledged their potential role in cell biology. Recently, several swine models of NF1 have been developed in hopes that these models will recapitulate the disorder in humans better than existing rodent models and thus offer a more translational platform for the development of novel therapies. In order to assess translatability of swine for NF1, we have characterized the tissue specific expression patterns of neurofibromin in newborn and adult swine. Tissue samples were collected from brain (frontal cortex, cerebellar vermis, amygdala, hippocampus, pituitary body, substantia nigra), nerves (brachial plexus, cranial nerve IV, optic nerve), heart (left ventricle apex), lung (lobe apex), muscle (superior trapezius), gastrointestinal tract (distal ileum, proximal large intestine), and skin (epithelium, adipose) of two‐day old (N=5) and eight‐month old (N=6) swine. The inclusion or exclusion of the four known alternatively spliced exons (ASEs) were determined by RT‐qPCR. Generally, inclusion of ASE 12 was relatively higher in cortical brain and muscle tissue, while inclusion of ASE 31, implicated in regulation of RAS activity, was relatively higher in myelin dense tissue. Inclusion of ASE 57 was significantly higher exclusively in muscle tissue. Interestingly, ASE 13 was not detected in any of the tissue and subsequent sequencing of the exon revealed a significant genomic variation compared to human: ASE 13 in swine contains an additional nucleotide that results in an early stop codon in neurofibromin transcription if this ASE was included. Overall neurofibromin expression was similar between age groups but expression of ASE 31 was overall lower in newborn tissue. Tissues from newborn swine also had lower expression of ASE 12 in brachial plexus, muscle, heart, and lung tissues. This study provides the first characterization of neurofibromin ASE expression in swine.Support or Funding InformationThe research was supported by the Biomedical &amp; Genomic Research Group Discretionary Fund (University of Wisconsin‐Madison), NF North Central, NF Network, and NF Team Foundation.

Influence of pre-pregnancy body mass index (p-BMI) and gestational weight gain (GWG) on DNA methylation and protein expression of obesogenic genes in umbilical vein.

Understanding the regulatory mechanisms that affect obesogenic genes expression in newborns is essential for early prevention efforts, but they remain unclear. Our study aimed to explore whether the maternal p-BMI and GWG were associated with regulatory single-locus DNA methylation in selected obesogenic genes. For this purpose, DNA methylation was assayed by Methylation-Sensitive High Resolution Melting (MS-HRM) technique and Sanger allele-bisulfite sequencing in fifty samples of umbilical vein to evaluate glucosamine-6-phosphate deaminase 2 (GNPDA2), peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC1α), and leptin receptor (LEPR) genes. Correlations between DNA methylation levels and indicators of maternal nutritional status were carried out. Western blotting was used to evaluate protein expression in extracts of the same samples. Results indicated that GNPDA2 and PGC1α genes have the same level of DNA methylation in all samples; however, a differential DNA methylation of LEPR gene promoter was found, correlating it with GWG and this correlation is unaffected by maternal age or unhealthy habits. Furthermore, leptin receptor (Lep-Rb) was upregulated in samples that showed the lowest levels of DNA methylation. This study highlights the association between poor GWG and adjustments on obesogenic genes expression in newborn tissues with potential consequences for development of obesity in the future.

A novel construct with biomechanical flexibility for articular cartilage regeneration

BackgroundAlthough tissue-engineered cartilage has been broadly studied, complete integration of regenerated cartilage with residual cartilage is still difficult for the inferior mechanical and biochemical feature of neocartilage. Chondrogenesis of mesenchymal stem cells can be induced by biophysical and biochemical factors.MethodsIn this study, autologous platelet-rich fibrin (PRF) membrane was used as a growth factor-rich scaffold that may facilitate differentiation of the transplanted bone marrow mesenchymal stem cells (BMSCs). At the same time, hydrostatic pressure was adopted for pre-adjustment of the seed cells before transplantation that may promote the mechanical flexibility of neocartilage.ResultsAn in vitro study showed that the feasible hydrostatic pressure stimulation substantially promoted the chondrogenic potential of in vitro-cultured BMSC/PRF construct. In vivo results revealed that at every time point, the newborn tissues were the most favorable in the pressure-pretreated BMSC/PRF transplant group. Besides, the transplantation of feasible hydrostatic pressure-pretreated construct by BMSC sheet fragments and PRF granules could obviously improve the integration between the regenerated cartilage and host cartilage milieu, and thereby achieve boundaryless repair between the neocartilage and residual host cartilage tissue in rabbit temporomandibular joints. It could be concluded that feasible hydrostatic pressure may effectively promote the proliferation and chondrogenic differentiation of BMSCs in a BMSC/PRF construct.ConclusionThis newly formed construct with biomechanical flexibility showed a superior capacity for cartilage regeneration by promoting the mechanical properties and integration of neocartilage.

Unique Innate Functions of Fetal Macrophage Populations

Abstract In fetal and newborn tissues, macrophages play key roles in generating the innate immune response to pathogens. Distinct macrophage populations arise from hematopoietic precursors in the embryonic yolk sac and fetal liver. Here we tested the hypothesis that fetal macrophage populations possess unique innate immune function based on their cellular origin. Innate immune activation following TLR agonist exposure was tested using FACS sorted fetal macrophage populations or total single cell suspension from fetal lung or fetal liver. TLR agonists increased inflammatory cytokine expression in CD11bHI fetal liver macrophages. However F4/80HI yolk sac derived macrophages failed to respond to TLR activation and expressed very low levels of both Il1b and Il6. Similar data were obtained with ELISA measurement of secreted IL-1beta and IL-6 protein. When total liver cells were treated with TLR agonists, again IL-1beta expression was restricted to CD11bHI macrophages. F4/80HI macrophages from the yolk sac were also less likely to phagocytose labeled E. coli. Expression of multiple TLRs was low or undetectable in F4/80HI macrophages, suggesting a unique overall functional program for this distinct macrophage population. Expression profiling showed similar levels of multiple macrophage signature genes in CD11bHI and F4/80HI cells. However expression of C1q family members and Igf1 were much higher in F4/80HI macrophages. Expression of these genes was more consistent with an anti-inflammatory phenotype and removal of damaged cells during developmental morphogenesis. These important mechanistic differences suggest important cellular and molecular strategies for treating neonatal inflammatory disease.

Effects of platelet-rich plasma combined with polylactic acid/polycaprolactone on healing of pig deep soft tissue defect caused by fragment injury

Objective: To investigate the effects of platelet-rich plasma (PRP) combined with polylactic acid/polycaprolactone (PLA/PCL) on healing of mininature pig deep soft tissue defect caused by fragment injury. Methods: Two male Bama miniature pigs with 11 to 12 months (the same below) were selected by lottery to prepare PRP. The other twenty-seven male Bama miniature pigs were used to reproduce deep soft tissue defect caused by high-explosive ammunition fragment injury on bilateral posterior femoral region. According to the random number table, 27 pigs were divided into control group, material group, and PRP+material group, with 9 pigs in each group. After debridement, wounds of pigs in material group and PRP+material group were filled with PLA/PCL and PLA/PCL+2 mL activated PRP, respectively. Pigs in each group received suture of full-thickness skin to close the wounds. The operative duration was recorded. The length and volume of wounds of pigs in the above groups were measured immediately after surgery. In 1, 2, and 4 weeks after surgery, 3 pigs in each group were sacrificed to collect femoral wounds tissue on two sides, and PLA/PCL were collected from wounds of pigs in material group and PRP+material group for general observation of wounds tissue and degradation of the material. In 2 and 4 weeks after surgery, wounds tissue was obtained to observe the histological changes by hematoxylin-eosin staining, and expressions of transforming growth factor β (TGF-β) and vascular endothelial growth factor (VEGF), and angiogenesis were determined by immunohistochemical method. In 1, 2, and 4 weeks after surgery, wounds tissue was collected to determine mRNA expressions of TGF-β and VEGF by real-time quantitative reverse transcription polymerase chain reaction. Data were processed with analysis of variance of factorial design, one-way analysis of variance, and least significant difference-t test. Results: (1) There were no significantly statistical differences in length and volume of the wounds of pigs among the three groups (F=0.336, 0.282, P>0.05). The operative duration in control group [(30.9±2.1)min] was significantly shorter than that of material group [(39.7±2.2)min] and PRP+material group[(40.0±2.6)min], t=-11.45, -11.88, P<0.01. (2) There were respectively 10, 7, and 5 wounds tissue with infection in pigs of control group, material group, and PRP+material group. In 1, 2, 4 weeks after surgery, all of the wounds tissue of pigs was infected in control group, while none of wounds tissue of pigs was infected in material group and PRP+material group. In pigs of material group and PRP+material group, materials and tissue were easily separated in 1 week after surgery; some materials were integrated with tissue and showed a tendency of degradation in 2 weeks after surgery; materials were completely embedded with tissue in 4 weeks after surgery. (3) In pigs of control group, erythrocytes and inflammatory cells infiltration in wounds tissue were observed in 2 weeks after surgery, and necrotic tissue and inflammatory cells infiltration in wounds tissue were still observed in 4 weeks after surgery. In pigs of material group and PRP+material group, a large number of erythrocytes and inflammatory cells infiltration were observed in 2 weeks after surgery. Compared with that of material group, wounds tissue of pigs in PRP+material group had no inflammatory cells infiltration in 4 weeks after surgery. (4) Protein expressions of TGF-β in fibroblasts and multinuclear macrophagocytes, VEGF in fibroblasts and vascular endothelial cells, and blood vessel formation in wounds tissue of pigs in PRP+material group were significantly more than those of pigs in control group and material group in 2 and 4 weeks after surgery. (5) The mRNA expression of TGF-β in wounds tissue of pigs in material group was significantly higher than that in control group in 4 weeks after surgery (t=-3.93, P<0.01). Compared with those of pigs in control group and material group, the mRNA expression of TGF-β in wounds tissue of pigs in PRP+material group was significantly increased at each time point (t=9.23, 13.81, 11.73, -7.51, -12.04, -7.80, P<0.01). The mRNA expression of VEGF in wounds tissue of pigs increased significantly in material group compared with that of pigs in control group in 4 weeks after surgery (t=-3.94, P<0.01). Compared with those of pigs in control group and material group, the mRNA expression of VEGF in wounds tissue increased significantly in wound tissue of pigs in PRP+material group at each time point (t=12.33, 3.95, 7.97, -11.36, -2.97, -4.04, P<0.01). Conclusions: PRP combined with PLA/PCL can accelerate wound healing of deep soft tissue defect of mininature pigs caused by fragment injury by providing physical scaffold for newborn tissue growth, promoting mRNA and protein expressions of TGF-β and VEGF.