Fetal Bone Research Articles

IL-4 promotes chondrogenesis of bone marrow mesenchymal stem cellsand blockade of IL-4Rα retards the endochondral ossification during rat embryonic bone development.

Interleukin-4 (IL-4)/IL-4 receptor alpha (IL-4Rα) signalling pathways play important roles in the complex process of bone formation and bone remodelling. However, whether IL-4/IL-4Rα participates in skeletogenesis during embryonic development is not completely understood. We used the anti-IL-4Rα monoclonal antibody (anti-IL-4Rα mAb) as a powerful investigational tool to evaluate the potential roles of IL-4/IL-4Rα in the chondrogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs) in vitro. Simultaneously, we explored the effect of IL-4/IL-4Rα on bone ossification during rat embryo-fetal development. In this study, we found that, compared to the control group, IL-4 can significantly promote the chondrogenic differentiation of BMSCs. Furthermore, following exposure to anti-IL-4Rα mAb in pregnant rats, unexpected phenomena were observed in fetal bone development, including non-ossification of the fetal sternum, an incomplete ossification centre in long bones and a reduced number of ossification points in digit (toe) bones. To further investigate the underlying mechanism of the phenotype, we studied the rat sternum as the target organ, starting from different time points of sternum development in the embryonic stage. The results indicated that the retardation mainly occurred in the middle and late stages of embryonic development. This retardation was characterized by the inhibition of the differentiation process of mesenchymal stem cells into chondrocytes, resulting in reduced angiogenesis near the ossification centre, failure of osteoblasts to invade the centre of the cartilage body with the blood vessels and delayed formation of the primary ossification centre (POC). Overall, our study demonstrated the significant function of IL-4/IL-4Rα in chondrogenic differentiation of BMSCs and bone ossification during embryo-fetal development.

The evolving hematopoietic niche during development.

Mammalian hematopoietic stem cells (HSCs) emerge from the hemogenic endothelium in the major embryonic arteries. HSCs undergo a complex journey first migrating to the fetal liver (FL) and from there to the fetal bone marrow (FBM), where they mostly remain during adult life. In this process, a pool of adult HSCs is produced, which sustains lifelong hematopoiesis. Multiple cellular components support HSC maturation and expansion and modulate their response to environmental and developmental cues. While the adult HSC niche has been extensively studied over the last two decades, the niches present in the major embryonic arteries, FL, FBM and perinatal bone marrow (BM) are poorly described. Recent investigations highlight important differences among FL, FBM and adult BM niches and emphasize the important role that inflammation, microbiota and hormonal factors play regulating HSCs and their niches. We provide a review on our current understanding of these important cellular microenvironments across ontogeny. We mainly focused on mice, as the most widely used research model, and, when possible, include relevant insights from other vertebrates including birds, zebrafish, and human. Developing a comprehensive picture on these processes is critical to understand the earliest origins of childhood leukemia and to achieve multiple goals in regenerative medicine, such as mimicking HSC development in vitro to produce HSCs for broad transplantation purposes in leukemia, following chemotherapy, bone marrow failure, and in HSC-based gene therapy.

Mineralization of equine proximal sesamoid bones precedes articular cartilage and fibrocartilaginous enthesis maturation in early postgestational development.

To describe the development and maturation of equine proximal sesamoid bones (PSBs) in fetuses and young horses using radiography, microcomputed (micro)-CT, and histology. A descriptive study. Forelimb PSBs from 12 equids ranging in age from 105 days of gestation to 540 days postgestation were evaluated. Radiography was used for preliminary assessment of metacarpophalangeal joint and PSB mineralization, and micro-CT imaging was performed to assess mineralized PSBs. Tissue volume, bone volume fraction, height, width, depth, trabecular thickness, and anisotropy were quantified from midplanar micro-CT sections in 3 dimensions. Midsagittal PSB histologic sections stained with H&E and Safranin O/Fast Green were used to determine the ratio of ossification center to cartilage template size and to describe the formation and development of the cartilage template, ossification center, spherical growth plate, articular cartilage, and entheses. Mineralization of equine PSBs is associated with cartilage canals and a spherical growth plate that undergoes endochondral ossification during the late gestation to early postgestational period. The apical, flexor, basilar, and articular ossification fronts demonstrate morphologic variability. Structural organization of the articular cartilage and entheses occurs concurrently with the development of an underlying plate of compact bone. At 540 days postgestation, the fibrocartilaginous entheses of the flexor cortex of the PSB had yet to mature. Equine PSBs mineralize predominantly by endochondral ossification during the late gestation to early postgestational period. Mineralization precedes maturation of the articular cartilage and fibrocartilaginous entheses. The postgestational maturation of the PSB and its surrounding tissues may predispose young horses to developing lesions at these sites, such as apical avulsion fractures, warranting further investigation.

Zinc and chitosan-enhanced β-tricalcium phosphate from calcined fetal bovine bone for mandible reconstruction.

Mandibular defects pose significant challenges in reconstructive surgery, and scaffold materials are increasingly recognized for their potential to address these challenges. Among various scaffold materials, Beta-tricalcium phosphate (β-TCP) is noted for its exceptional osteogenic properties. However, improvements in its biodegradation rate and mechanical strength are essential for optimal performance. In this study, we developed a novel β-TCP-based scaffold, CFBB, by calcining fetal bovine cancellous bone. To enhance its properties, we modified CFBB with Chitosan (CS) and Zinc (Zn), creating three additional scaffold materials: CFBB/CS, CFBB/Zn2+, and CFBB/Zn2+/CS. We conducted comprehensive assessments of their physicochemical and morphological properties, degradation rates, biocompatibility, osteogenic ability, new bone formation, and neovascularization both in vitro and in vivo. Our findings revealed that all four materials were biocompatible and safe for use. The modifications with CS and Zn2+ significantly improved the mechanical strength, osteogenic, and angiogenic properties of CFBB, while concurrently decelerating its resorption rate. Among the tested materials, CFBB/Zn2+/CS demonstrated superior performance in promoting bone regeneration and vascularization, making it a particularly promising candidate for mandibular reconstruction. The CFBB/Zn2+/CS scaffold material, with its enhanced mechanical, osteogenic, and angiogenic properties, and a controlled resorption rate, emerges as a highly effective alternative for the repair of oral mandible defects. This study underscores the potential of combining multiple bioactive agents in scaffold materials to improve their functionality for specific clinical applications in bone tissue engineering.

Single-cell multi-omics map of human fetal blood in Down syndrome

Down syndrome predisposes individuals to haematological abnormalities, such as increased number of erythrocytes and leukaemia in a process that is initiated before birth and is not entirely understood1–3. Here, to understand dysregulated haematopoiesis in Down syndrome, we integrated single-cell transcriptomics of over 1.1 million cells with chromatin accessibility and spatial transcriptomics datasets using human fetal liver and bone marrow samples from 3 fetuses with disomy and 15 fetuses with trisomy. We found that differences in gene expression in Down syndrome were dependent on both cell type and environment. Furthermore, we found multiple lines of evidence that haematopoietic stem cells (HSCs) in Down syndrome are ‘primed’ to differentiate. We subsequently established a Down syndrome-specific map linking non-coding elements to genes in disomic and trisomic HSCs using 10X multiome data. By integrating this map with genetic variants associated with blood cell counts, we discovered that trisomy restructured regulatory interactions to dysregulate enhancer activity and gene expression critical to erythroid lineage differentiation. Furthermore, as mutations in Down syndrome display a signature of oxidative stress4,5, we validated both increased mitochondrial mass and oxidative stress in Down syndrome, and observed that these mutations preferentially fell into regulatory regions of expressed genes in HSCs. Together, our single-cell, multi-omic resource provides a high-resolution molecular map of fetal haematopoiesis in Down syndrome and indicates significant regulatory restructuring giving rise to co-occurring haematological conditions.

3D printing of realistic body phantoms: Comparison of measured and simulated organ doses on the example of a CT scan on a pregnant woman

AbstractBackgroundMedical examinations or treatment of pregnant women using ionizing radiation are sometimes unavoidable. In such cases, the risk of harm to the embryo and fetus after exposure to ionizing radiation must be carefully estimated. However, no commercially available anthropomorphic body phantoms of pregnant women are available for dose measurements. A promising possibility for the production of body phantoms for patient groups that are not adequately represented by the phantoms of reference persons is 3D printing. However, this approach is still in the evaluation phase.PurposeTo print the abdomen of a woman in the late stage of pregnancy and compare the dose distribution measured using thermoluminescence dosimeters (TLDs) in the printed phantom for two different computed tomography (CT) protocols with the corresponding results of Monte Carlo simulations on voxel models of the pregnant woman.Materials and methodsThe physical phantom was produced through multi‐material extrusion printing using different print materials identified in previous studies to simulate homogeneous soft tissues and the mean compositions of maternal and fetal bones. The 3D printed abdomen was combined with a conventionally produced anthropomorphic female phantom to obtain a whole‐body phantom of a pregnant woman. Dose values resulting from two different CT scans acquired at tube voltages of 80 and 120 kV were measured using TLDs positioned in the physical phantom and cross‐validated with the results of Monte Carlo simulations performed for two different voxel models. The first was a voxelized model of the produced phantom itself and the second a realistic digital model of a pregnant woman. Representative CT values of the materials used in the printed phantom were determined from the acquired CT images.ResultsThe CT values of maternal and fetal tissue structures in the phantom are comparable to CT values of real human tissues. The difference between most organ doses measured in the 3D printed phantom and simulated in the voxel models was below 20% and equivalent within the measurement uncertainties. Only the dose to the fetal head was up to 50% higher and not equivalent for the realistic model and the 80 kV‐protocol. As expected, the agreement was better for the voxelized than for the realistic model. For both models a slight energy dependence was observed, with larger deviations for the 80‐kV protocol especially for organs located in the pelvic region.ConclusionIndividualized physical body phantoms, such as that of a pregnant woman, can be produced using 3D printing. The good agreement between measured and simulated doses to the fetus cross‐validates both dosimetric methods. Therefore, this study demonstrates the suitability of 3D printing phantoms for patients not adequately represented by commercially available body phantoms of reference persons.

Fetal bone engraftment reconstitutes the immune system in pigs with severe combined immunodeficiency.

Genetic modification of genes such as recombination activating gene 2 (RAG2) or interleukin-2 receptor-γ (IL2RG) results in pigs exhibiting severe combined immunodeficiency (SCID). Pigs presenting a SCID phenotype are important animal models that can be used to establish xenografts and to study immune system development and various immune-related pathologies. However, due to their immunocompromised nature, SCID pigs have shortened lifespans and are notoriously difficult to maintain. The failure-to-thrive phenotype makes the establishment of a breeding population of RAG2/IL2RG double-knockout pigs virtually impossible. Here, to overcome this limitation, we investigated whether reconstituting the immune system of SCID piglets with a fetal bone allograft would extend their lifespan. Following intramuscular transplantation, allografts gave rise to lymphocytes expressing T cell (CD3, CD4 and CD8), B cell (CD79α) and natural killer cell (CD335) lineage markers, which were detected in circulation as well in the spleen, liver, bone marrow and thymic tissues. The presence of lymphocytes indicates broad engraftment of donor cells in the recipient SCID pigs. Unlike unreconstituted SCID pigs, the engrafted animals thrived and reached puberty under standard housing conditions. This study demonstrates a novel method to extend the survival of SCID pigs, which may improve the availability and use of SCID pigs as a biomedical animal model.

The ontogeny of human fetal trabecular bone architecture occurs in a limb-specific manner

Gestational growth and development of bone is an understudied process compared to soft tissues and has implications for lifelong health. This study investigated growth and development of human fetal limb bone trabecular architecture using 3D digital histomorphometry of microcomputed tomography data from the femora and humeri of 35 skeletons (17 female and 18 male) with gestational ages between 4 and 9 months. Ontogenetic data revealed: (i) fetal trabecular architecture is similar between sexes; (ii) the proximal femoral metaphysis is physically larger, with thicker trabeculae and greater bone volume fraction relative to the humerus, but other aspects of trabecular architecture are similar between the bones; (iii) between 4 and 9 months gestation there is no apparent sexual or limb dimorphism in patterns of growth, but the size of the humerus and femur diverges early in development. Additionally, both bones exhibit significant increases in mean trabecular thickness (and for the femur alone, bone volume fraction) but minimal trabecular reorganisation (i.e., no significant changes in degree of anisotropy, connectivity density, or fractal dimension). Overall, these data suggest that in contrast to data from the axial skeleton, prenatal growth of long bones in the limbs is characterised by size increase, without major reorganizational changes in trabecular architecture.

MiR-322-5p is involved in regulating chondrocyte proliferation and differentiation in offspring’s growth plate of maternal gestational diabetes

Pregestational diabetes mellitus (PGDM) has an impact on fetal bone formation, but the underlying mechanism is still obscure. Although miRNAs have been extensively investigated throughout bone formation, their effects on fetal bone development caused by PGDM still need clarification. This study intends to examine the mechanism by which hyperglycemia impairs the bone formation of offspring via miR-322-5p (miR-322). In this study, miR-322 was selected by systemically screening utilizing bioinformatics and subsequent validation experiments. Using streptozotocin (STZ)-induced diabetic mice and ATDC5 cell lines, we found that miR-322 was abundantly expressed in the proliferative and hypertrophic zones of the growth plate, and its expression pattern was disturbed in the presence of hyperglycemia, suggesting that miR-322 is involved in the chondrocyte proliferation and differentiation in absence/presence of hyperglycemia. This observation was proved by manipulating miR-322 expression in ATDC5 cells by transfecting mimic and inhibitor of miR-322. Furthermore, Adamts5, Col12a1, and Cbx6 were identified as the potential target genes of miR-322, verified by the co-transfection of miR-322 inhibitor and the siRNAs, respectively. The evaluation criteria are the chondrocyte proliferation and differentiation and their relevant key gene expressions (proliferation: Sox9 and PthIh; differentiation: Runx2 and Col10a1) after manipulating the gene expressions in ATDC5 cells. This study revealed the regulative role miR-322 on chondrocyte proliferation and differentiation of growth plate by targeting Adamts5, Col12a1, and Cbx6 in hyperglycemia during pregnancy. This translational potential represents a promising avenue for advancing our understanding of bone-related complications in diabetic pregnancy and mitigating bone deficiencies in diabetic pregnant individuals, improving maternal and fetal outcomes.

A new insight for investigating the prenatal and postnatal ossification centers of pelvic and femur bones in white New Zealand rabbits (Oryctolagus cuniculus) using 3D CT, double stain technique, and morphometry

BackgroundThe ossification centers in rabbit limbs are related to fetal age and bone maturation. Objective: To address the limited studies on ossification in the hind limbs of New Zealand rabbits, we investigated the prenatal and postnatal development of the pelvic and femur bones. MethodsDouble staining with Alcian Blue and Alizarin Red, computed tomography (CT), and 3D reconstruction were employed to visualize and analyze ossification centers in detail. ResultsUsing double staining, we observed these patterns: At prenatal days 18 and 21, ossification centers appeared in the ilium. By prenatal days 23 and 25, ossification began in the ischium. On postnatal day 1, ilium ossification centers spread across most of the ilium wings, except for the iliac crest, and new centers appeared in the pubis and cotyloid bones. Most bones had ossified by the third week and one month postnatal, except for the iliac crest and ischial tuberosity. At 1.5 months, both were fully ossified. On day 18 post coitum, an ossification center was visible in the middle of the femur shaft. By day 28 post coitum, ossification extended through the shaft, and postnatally, new ossification spots appeared at the extremities by day one and week one. By the third week, complete ossification of the femur head, lesser trochanter, third trochanter, medial condyle, and lateral condyle was observed. At 1.5 months, the entire proximal extremity was ossified. Conclusion3D CT provided clear imaging of ossification progression in the pelvic and femur bones. This study enhances our understanding of vertebrate skeletal development.

Growth and mineralization of fetal mouse long bones under microgravity and daily 1 g gravity exposure

In a previous Space Shuttle/Spacelab experiment (STS-42), we observed direct responses of isolated fetal mouse long bones to near weightlessness. This paper aimed to verify those results and study the effects of daily 1×g exposure during microgravity on the growth and mineralization of these bones. Two experiments were conducted: one on an American Space Shuttle mission (IML-2 on STS-65) and another on a Russian Bio-Cosmos flight (Bion-10 on Cosmos-2229). Despite differences in hardware, both used 17-day-old fetal mouse metatarsals cultured for 4 days. Results showed reduced proteoglycan content under microgravity compared to 1×g conditions, with no main differences in other cellular structures. While the overall metatarsal length was unaffected, the length increase of the mineralized diaphysis was significantly reduced under microgravity. Daily 1×g exposure for at least 6 h abolished the microgravity-induced reduction in cartilage mineralization, indicating the need for long-duration exposure to 1×g as an in-flight countermeasure using artificial gravity.

Correlation of Gestational Age with Anterior-Posterior Thigh Diameter Measured by Two-Dimensional Sonography

Background: Anterior-posterior thigh diameter measurement via two-dimensional sonography (2D-US) correlates with fetal gestational age and reflects the growth and development of fetal thigh muscles and bones. This measurement, alongside estimated gestational age, offers valuable insights into fetal well-being and growth during pregnancy. This study aimed to evaluate the correlation of gestational age with anterior-posterior thigh diameter measured by two-dimensional sonography and estimated gestational age. Methods: This cross-sectional study was conducted at the Department of Radiology and Imaging, Mymensingh Medical College & Hospital, Mymensingh, Bangladesh from January 2010 to January 2012. As the study subjects, a total of 250 healthy women between the 24th and 38th weeks of normal pregnancy were enrolled by using a purposive sampling technique. For data analysis, SPSS version 23.0 was used. Results: The mean age of the participants was 24.79 ± 4.71 years with a range from 18 to 35 years. More than one-third (34.0%) of the women were in the age range of 21-25 years. The mean gestational age of all participants was 30.93 ± 4.32 weeks. Primigravida accounted for more than half (56.4%) of them. A statistically significant positive correlation was observed between fetal anterior-posterior thigh diameter and gestational age (p<0.001. Conclusion: There is a statistically significant positive correlation between fetal anterior-posterior thigh diameter and gestational age.

Global Transcriptomic and Characteristics Comparisons between Mouse Fetal Liver and Bone Marrow Definitive Erythropoiesis.

Erythropoiesis occurs first in the yolk sac as a transit "primitive" form, then is gradually replaced by the "definitive" form in the fetal liver (FL) during fetal development and in the bone marrow (BM) postnatally. While it is well known that differences exist between primitive and definitive erythropoiesis, the similarities and differences between FL and BM definitive erythropoiesis have not been studied. Here we performed comprehensive comparisons of erythroid progenitors and precursors at all maturational stages sorted from E16.5 FL and adult BM. We found that FL cells at all maturational stages were larger than their BM counterparts. We further found that FL BFU-E cells divided at a faster rate and underwent more cell divisions than BM BFU-E. Transcriptome comparison revealed that genes with increased expression in FL BFU-Es were enriched in cell division. Interestingly, the expression levels of glucocorticoid receptor Nr3c1, Myc and Myc downstream target Ccna2 were significantly higher in FL BFU-Es, indicating the role of the Nr3c1-Myc-Ccna2 axis in the enhanced proliferation/cell division of FL BFU-E cells. At the CFU-E stage, the expression of genes associated with hemoglobin biosynthesis were much higher in FL CFU-Es, indicating more hemoglobin production. During terminal erythropoiesis, overall temporal patterns in gene expression were conserved between the FL and BM. While biological processes related to translation, the tricarboxylic acid cycle and hypoxia response were upregulated in FL erythroblasts, those related to antiviral signal pathway were upregulated in BM erythroblasts. Our findings uncovered previously unrecognized differences between FL and BM definitive erythropoiesis and provide novel insights into erythropoiesis.

Evaluation of anatomical and radiological measurements of the proximal phalanx of the horse

When performing osteosynthesis by screw fixation, it is important to carry out operational planning (choosing the screw of the optimal length and diameter). Choosing the optimal osteosynthesis tactics and the correct screw size allows you to stimulate regeneration and prevent possible complications. The X-ray examination method is included in the mandatory criterion of preoperative examination, which allows you to determine the size of the deformation and bone parameters. The present study is based on the hypothesis that the lifetime X-ray measurement of the bone of the proximal phalanx of a horse (longitudinal and transverse dimensions of the proximal and distal epiphysis and diaphysis, bone length), allows us to justify the size and technique of fixing the screw in the treatment of fractures. Morphometric studies included macroscopic dissection; morphometry of bone preparations; Xray measurement of the putus bone; documentation of the data obtained, photographing. A morphometric study was performed on 20 bone preparations (10 thoracic and 10 pelvic) and 40 X-ray images (20 in lateromedial and 20 in dorso-palmar/plantar projections) of the proximal phalanx of the horse. As a result of the work performed, it was found that the variability of parameters (longitudinal and transverse size of the proximal epiphysis, longitudinal and transverse size of the distal epiphysis; the longitudinal and transverse size of the diaphysis, bone length) of the fetal bone of the thoracic and pelvic limbs in horses for average values has no significant deviations (up to 1%), while the differences between the minimum and maximum values range from 20.0% to 66.6% for values obtained by anatomical method; from 12.2% and 33.3% for the Xray method.

Transrectal ultrasonographic assessment of the fetal proximal phalanx: A new tool to assess fetal age and bone development in horses

Fetal age in Quarter Horses can be predicted within 2 weeks from 100- to 200- days of gestation using femur length, biparietal diameter (cranium diameter) and eye approximated volume. However, as pregnancy advances, the femur and cranium become too large to be imaged in their entirety using ultrasound and the corresponding biometric parameters can no longer be measured. In this longitudinal study, the proximal phalanx (P1) was evaluated as a novel biometric parameter for late gestation to predict fetal age and bone maturation. Transrectal ultrasound was performed in ten pregnant mares with known ovulation dates, every two weeks from 240- days of gestation until parturition. P1 was imaged in 69 % of the examinations. Inability to image P1 was due to obstructive positioning such as carpal or fetlock flexion, or posterior presentation of the fetus. Advancing fetal age did not affect visibility of P1. P1 length correlated significantly with days of gestation and a correlation equation was established: y = 0.3837x -69.55 where y is the predicted value of P1 length and x is the day of gestation (with day 0 being the day of ovulation). When P1 length was equal to or larger than the width of the ultrasound image (52.5 mm), 90 % of mares (9/10) were above 300- days of gestation. Ossification of the proximal and distal epiphysis of P1 typically appeared between 277- and 303 -days of gestation (mean: 288 days). The proximal epiphysis did not close before parturition whereas the distal one closed between 306- and 333-days of gestation (mean: 320 days). P1 epiphyseal appearance and closure occurred chronologically reflecting bone maturation. Radiographic findings at birth and prenatal ultrasound findings were in agreement, apart from timing of P1 distal epiphyseal closure. In conclusion, P1 length can be used as a new fetal biometric parameter to assess fetal age and growth after 240- days of gestation. The knowledge of P1 bone maturation process in utero as a marker for fetal bone development, may also be valuable in clinical decision-making when considering inducing parturition in the mare.

Quantitative study of the ossification centers of the body of sphenoid bone in the human fetus

The aim of the present study was to examine the growth dynamics of the two ossification centers of the body of sphenoid bone in the human fetus, based on their linear, planar and volumetric parameters. The examinations were carried out on 37 human fetuses of both sexes aged 18–30 weeks of gestation, which had been preserved in 10% neutral formalin solution. Using CT, digital image analysis software, 3D reconstruction and statistical methods, we evaluated the size of the presphenoid and postsphenoid ossification centers. The presphenoid ossification center grew proportionately in sagittal diameter, projection surface area and volume, and logarithmically in transverse diameter. The postsphenoid ossification center increased logarithmically in sagittal diameter, transverse diameter and projection surface area, while its volumetric growth followed proportionately. The numerical findings of the presphenoid and postsphenoid ossification centers may be considered age-specific reference values of potential relevance in monitoring the normal fetal growth and screening for congenital disorders in the fetus. The obtained results may contribute to a better understanding of the growing fetal skeleton, bringing new numerical information regarding its diagnosis and development.

Fetal ultrasound anatomy and morphometric parameters of the tibia

Ultrasound screening is one of the most informative methods of visualizing fetal development during pregnancy. During standard ultrasound examinations, the length of the femur of the fetus is measured. However, if any skeletal dysplasia or intrauterine growth retardation is suspected, additional measurement of other long tubular bones should be performed. The aim of the work was to investigate the morphometric parameters of the lengths of the right and left tibia bones and to establish correlations between them and the lengths of the corresponding fibula, lower limb and crown rump length of the fetus during the fetal period of human ontogenesis. Intravital ultrasound examination of the leg bones of 38 human fetuses was performed at the medical center "YUZKO MEDICAL CENTER" in accordance with the cooperation agreement. Built-in capabilities of MS Excel were used for statistical calculations. The established data on the fetal morphometry of tibial lengths indicate relatively uniform growth of the right and left tibias during the fetal period of human ontogenesis: the length of the right tibia increases from 24.77±2.25 mm to 65.32±2.20 mm, and the length of the left tibia increases from 25.19±1.63 mm to 65.78±1.81 mm. The analysis of correlations between the length of the right and left tibia bones, the length of the fibula bones, the length of the lower limbs and the crown rump length in human fetuses aged from 4 to 10 months of intrauterine development allowed to reveal reliable strong and medium strength reliable and unreliable, mostly direct, connections in all age groups. The obtained morphometric parameters of tibia bones in fetuses of different gestational periods are important additional dimensions of fetobiometry, which can be used if a detailed assessment of the state of fetal development is necessary and in case of suspicion of congenital anomalies.

Histological Observations of the Developing Human Fetal Humerus at Different Trimesters of Pregnancy

Background: Although femur length is the preferred method for fetal age assessment, the humerus is sometimes the right choice, especially in the second half of pregnancy. Fetal bone development significantly impacts adult bone quality and senescent bone disorders, including osteoporosis. Purpose of the study: The present study aimed to ascertain the histological changes in the human fetal humerus across the three trimesters. After the institutional ethical committee clearance and parent consent, the study was carried out on stillborn or medically terminated human fetuses from the 10th to the 32nd week of intrauterine life. Results: First trimester: The primary bone collar appeared with the primary ossification center, marked by more vascular invasion and a pool of mesenchymal cells. Trabeculations begin from the bony collar and insignificant periosteum. Second trimester: The changes showed longitudinal growth of periosteal bone towards the proximal and distal ends of the growing bone. Growth plate with distinguished zones and gradual fusion of epiphysis with growth plate were observed. Third trimester: Trabeculation number and thickness increase with calcification. Towards the third trimester, the marrow cavity with increased and prominent trabeculations is consistent. Conclusion: Understanding normal microstructural and cellular events chronologically is an ideal platform for future studies to develop cell-based or cell-targeted therapies for adult bone disorders, traumatic bone injuries, or bone engineering. KEYWORDS: Histology, Developing Bone, Humerus, Fetuses, Embryology.

The “toxic window” of amoxicillin exposure during pregnancy on long bone development in fetal mice

AimsAmoxicillin is a broad-spectrum beta-lactam antibiotic used to treat infectious diseases in pregnant women. Studies have shown that prenatal amoxicillin exposure (PAmE) has developmental toxicity on fetal development. However, the effect of PAmE on long bone development has not been reported. This study aimed to investigate the “toxic window” of PAmE on long bone development and explore its possible mechanism in fetal mice. Materials and methodsPregnant mice were administered amoxicillin by gavage at different stages (gestational day (GD)10–12 and GD16–18), different doses (150 and 300 mg/kg·d) and different courses (single and multiple courses). Fetal femurs were collected at GD18 and bone development related indicators were detected. Key findingsThe results showed that PAmE significantly reduced the length of the femur and primary ossification center of fetal mice, and inhibited the development of fetal growth plate. Meanwhile, PAmE inhibited the development of bone marrow mesenchymal stem cells, osteoclasts and endothelial cells in fetal long bone. Further, we found the fetal long bone developmental toxicity induced by PAmE was most significant at late-pregnancy (GD16–18), high dose (300 mg/kg·d) and multiple-course group. Besides, PAmE inhibited the expression of Wnt/β-catenin signaling pathway in fetal long bone. The β-catenin mRNA expression was significantly positively correlated with the development indexes of fetal long bone. SignificancePAmE has toxic effects on long bone development, and there was an obvious “toxic window” of PAmE on the long bone development in fetal mice. The Wnt/β-catenin signaling pathway may mediate PAmE-induced fetal long bone development inhibition.

Prenatal diagnosis of fetuses with absent/hypoplastic nasal bone in second-trimester using chromosomal microarray analysis.

Pathogenic copy number variants (pCNVs) are associated with fetal ultrasound anomalies, which can be efficiently identified through chromosomal microarray analysis (CMA). The primary objective of the present study was to enhance understanding of the genotype-phenotype correlation in fetuses exhibiting absent or hypoplastic nasal bones using CMA. Enrolled in the present study were 94 cases of fetuses with absent/hypoplastic nasal bone, which were divided into an isolated absent/hypoplastic nasal bone group (n = 49) and a non-isolated group (n = 45). All pregnant women enrolled in the study underwent karyotype analysis and CMA to assess chromosomal abnormalities in the fetuses. Karyotype analysis and CMA detection were successfully performed in all cases. The results of karyotype and CMA indicate the presence of 11 cases of chromosome aneuploidy, with trisomy 21 being the most prevalent among them. A small supernumerary marker chromosome (sSMC) detected by karyotype analysis was further interpreted as a pCNV by CMA. Additionally, CMA detection elicited three cases of pCNVs, despite normal findings in their karyotype analysis results. Among them, one case of Roche translocation was identified to be a UPD in chromosome 15 with a low proportion of trisomy 15. Further, a significant difference in the detection rate of pCNVs was observed between non-isolated and isolated absent/hypoplastic nasal bone (24.44% vs. 8.16%, p < .05). The present study enhances the utility of CMA in diagnosing the etiology of absent or hypoplastic nasal bone in fetuses. Further, isolated cases of absent or hypoplastic nasal bone strongly suggest the presence of chromosomal abnormalities, necessitating genetic evaluation through CMA.