Ossification Centers Research Articles

Prenatal growth patterns of the upper jaw complex with implications for laryngeal echolocation in bats.

Craniofacial morphology is extremely diversified within bat phylogeny, however growth and development of the palate in bats remains unstudied. The formation of both midline and bilateral orofacial clefts in laryngeally echolocating bats, morphologically similar to the syndromic and non-syndromic cleft palate in humans, are not well understood. Developmental series of prenatal samples (n = 128) and adults (n = 10) of eight bat species (two pteropodids, four rhinolophoids, and two yangochiropterans), and two non-bat mammals (Mus musculus and Erinaceus amurensis), were CT-scanned and cranial bones forming the upper jaw complex were three-dimensionally visualised to assess whether differences in palate development can be observed across bat phylogeny. Volumetric data of bones composing the upper jaw complex were measured to quantify palate growth. The premaxilla is relatively reduced in bats compared to other mammals and its shape is heterogeneous depending on the presence and type of orofacial cleft across bat phylogeny. The palatine process of premaxillary bones is lacking in pteropodids and yangochiropterans, whereas the premaxilla is a mobile structure which is only in contact caudally with the maxilla by a fibrous membrane or suture in rhinolophoids. In all bats, maxillary bones progressively extend caudally and palatine bones, in some cases split into three branches, extend caudally so that they are completely fused to another one medially prior to the birth. Ossification of the vomer and fusion of the maxillary and palatine bones occur earlier in rhinolophoids than in pteropodids and yangochiropterans. The vomer ossifies bilaterally from two different ossification centres in yangochiropterans, which is uncommon in other bats and non-bat mammals. Analysis of ontogenetic allometric trajectories of the upper jaw complex revealed faster development of maxillary, vomer, and palatine bones in yangochiropterans compared to other bats, especially rhinolophoids. Ancestral state reconstruction revealed that yangochiropterans have a higher magnitude of change in ossification rate compared to other bats and E. amurensis a lower magnitude compared to M. musculus and bats. This study provides new evidence of heterochronic shifts in craniofacial development and growth across bat phylogeny that can improve understanding of the developmental differences characterising nasal and oral emission strategies.

Profile of Medico-legal Age Estimation Cases in a Tertiary Hospital in Kathmandu

Introduction: Medico-legal age estimation is the estimation of chronological age of an individual which is based upon analysis of physical development of the body, development of secondary sexual traits, tooth eruption, appearance of ossification centers of bones, and the degree of epiphyseal union. Chronological age of the subject whether a minor (less than 18 years) or an adult determines the fate of the case in the court of law. Objective: The present study revealed varied profiles of the perpetrator and the victim who came for medico-legal age estimation. Methods: This is a retrospective, descriptive cross-sectional study conducted in the Department of Forensic Medicine, Maharajgunj Medical Campus, Institute of Medicine (IoM). The data of medico-legal age estimation of 5 years period was analyzed using the statistical software SPSS 20. Results: Total of 313 individuals were studied. Majority of the individuals were within the age group of 16-17 years. Males (70.3 %) outnumbered females (29.7 %). Theft (31.9 %) was common among subjects, followed by sexual assault (18.2 %) and majority of subjects (59.7%) came from Province 3. Sixty percent of the subject’s alleged age, matched the estimated ages. Conclusion: This study shows the status of children in crime which demands attention of concerned authority. Males were significantly involved in theft and females were victims of sexual assault. Forty percent of the alleged age given by the subject differed from the estimated age, to change the fate of the case. These results could greatly benefit the judiciary system of Nepal.

Unique bone histology of modern giant salamanders: a study on humeri and femora of Andrias spp.

The osteohistology of Andrias spp. is a pivotal analogue for large fossil non-amniotes (e.g., Temnospondyli), and the endangered status of this taxon underlines the importance of gathering information on its growth. We here present the first osteohistological study by petrographic thin sections of an ontogenetic series of humeri and femora of eight individuals of varying sizes (28.5–104 cm) and ages (2.5–32 years) of Andrias japonicus from the Hiroshima City Asa Zoological Park, Japan. In addition, two individuals of A. cf. davidianus of unknown age but of different size (62 cm and 94 cm) were studied. All samples of Andrias spp. show a primary avascular periosteal cortex made of parallel-fibred tissue around the ossification center in the petrographic thin sections. Mainly in small individuals, the fibers forming this tissue are very coarse and loosely organized. With increasing size and age, the coarse tissue is irregularly intermixed and later replaced with finer and better organized fibers. This histologic change is accompanied by a change from diffuse annuli in the inner cortex to distinct lines of arrested growth (LAGs) in the outer cortex. We interpret these changes in tissue and the appearance of distinct growth marks as indicating the onset of active reproduction. The lack of primary vascularization around the ossification center in our Andrias spp. sample is striking and contradicts other observations. Vascularity may be prone to plasticity and further studies are necessary. We hypothesize that the large osteocyte lacunae and the dense networks of canaliculi observed in our sample may have nourished the tissue instead of primary vascular canals. We measured the size of osteocyte lacunae of Andrias spp. in comparison to other Lissamphibia, and found them to be significantly larger throughout ontogeny. The periosteal cortex contains a high amount of thick Sharpey’s fibers all around the midshaft cross sections. The two samples of Andrias cf. davidianus show tissue and growth mark distribution similar to that observed in A. japonicus. However, the large individual of A. cf. davidianus differed in its extremely osteosclerotic condition and the retention of a small layer of calcified cartilage in the endosteal region of the femur. It remains unclear whether these differences are related to plasticity, taxonomy, sex, exogenous factors, or attributable to a regenerated but fully regrown leg. Although the present study is based on zoo-kept and not wild, animals, it yields important insights into osteohistological plasticity and growth patterns in giant salamanders.

Comparison Between Ultrasonography and Radiography in the Detection of Epiphyseal Ossification Centers of the Knee in Infants With Permanent Congenital Hypothyroidism.

To demonstrate the usefulness of ultrasonography in detecting knee ossification centers in infants with permanent congenital hypothyroidism (PCH). From 2011 to 2021, all infants with PCH referred for thyroid ultrasound also underwent left knee ultrasound and radiography on the same day. Knee radiographs were compared with knee sonograms. Two pediatric radiologists reviewed the consensus knee radiographs and sonograms to identify femoral and tibial epiphyseal ossification centers (presence/absence). The concordance between ultrasonography and radiography was assessed. Another radiologist conducted a second late review to evaluate interobserver agreement. We identified 125 patients (65 girls, 60 boys) with a mean age of 24 days (5 days-5 months). On scintigraphy, the thyroid was in place in 66.4%, ectopic in 24%, and absent in 9.6% of patients. The femoral center was observed in 108 patients (86.4%) via sonography and 106 patients (84.8%) via radiography. The tibial center was observed in 84 patients (67.2%) via sonography and radiography. Both femoral and tibial centers were present on sonography and radiography in 84 patients (67.2%). A single nucleus was present in 24 patients (19.2%) on sonography and 22 patients (17.6%) on radiography; it corresponded to the femoral center in all patients. The concordance between ultrasonography and radiography was 99% and 100%, respectively, for the detection of the femoral and tibial centers. Interobserver agreement was substantial to almost perfect for both ultrasonography and radiography. Ultrasonography is as effective as radiography in detecting knee ossification centers in PCH. It can be performed at the same time as thyroid examination, in place of radiography.

Digital image analysis of vertebral body L4 and its ossification center in the human fetus.

Using a Siemens-Biograph 128 mCT camera the morphometric analysis of the L4 vertebral body and its ossification center were done in 55human fetuses aged 17 to 30weeks. No sex differences were found. The mean height, transverse and sagittal diameters of L4 vertebral body followed the logarithmic functions: y = -11.797+ 5.208 × ln(age) ± 0.372, y = -23.462 + 9.428 × ln(age) ± 0.702, y = 2.770 + 13.521 × ln(age) ± 1.722, respectively. The mean cross-sectional area of L4 vertebral body followed the linear function: y = -30.683 + 1.976 × age ± 2.701. The mean volume of L4 vertebral body followed the second-degree polynomial function: y = -93.983+ 0.385 × (age)² ± 23.707. The mean transverse and sagittal diameters of the ossification center of L4 vertebral body followed the natural logarithmic function: y = -27.106 + 10.178 × ln(age) ± 0.769 and y = -13.345 + 5.458 × ln(age) ± 0.424, respectively. The mean cross-sectional area and the volume of the ossification center of L4 vertebral body followed the linear function: y = -30.683 + 1.976 × age ± 2.701 and y = -43.214 + 2.760 × age ± 4.085, respectively.

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.

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.

A Osteocondrose e a Traumatologia do Desporto

Osteochondrosis is defined as a failure in the endochondral ossification process. There is no consensus on the etiology, with multiple factors involved. The initial events in the pathogenesis remain uncertain, but evidence points to ischemic necrosis of the ossification center, located in the bone epiphysis. Each osteochondrosis has a homonymous clinical diagnosis assigned based on location. The most affected areas include the hip (Legg-Calvé-Perthes disease), knee (Osgood-Schlatter disease, Sinding-Larsen-Johansson disease), foot (Sever’s disease), elbow (Panner’s disease) and spine (Scheuermann’s disease). The most common clinical manifestations are mechanical pain and functional limitation during or after physical activity. Many injury prevention strategies involve educating young people, coaches, family members and monitoring symptoms. Early diagnosis and a structured approach allow, in most cases, clinical resolution and avoid long-term sequelae. Osteochondrosis was first described by König in 1887 and, despite more than a century of studies, there are still many challenging questions to elucidate to better understand this pathology.

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.

Fibronectin isoforms promote postnatal skeletal development

Fibronectin (FN) is a ubiquitous extracellular matrix glycoprotein essential for the development of various tissues. Mutations in FN cause a unique form of spondylometaphyseal dysplasia, emphasizing its importance in cartilage and bone development. However, the relevance and functional role of FN during skeletal development has remained elusive. To address these aspects, we have generated conditional knockout mouse models targeting the cellular FN isoform in cartilage (cFNKO), the plasma FN isoform in hepatocytes (pFNKO), and both isoforms together in a double knockout (FNdKO). We used these mice to determine the relevance of the two principal FN isoforms in skeletal development from postnatal day one to the adult stage at two months.We identified a distinct topological FN deposition pattern in the mouse limb during different gestational and postnatal skeletal development phases, with prominent levels at the resting and hypertrophic chondrocyte zones and in the trabecular bone. Cartilage-specific cFN emerged as the predominant isoform in the growth plate, whereas circulating pFN remained excluded from the growth plate and confined to the primary and secondary ossification centers. Deleting either isoform independently (cFNKO or pFNKO) yielded only relatively subtle changes in the analyzed skeletal parameters. However, the double knockout of cFN in the growth plate and pFN in the circulation of the FNdKO mice significantly reduced postnatal body weight, body length, and bone length. Micro-CT analysis of the adult bone microarchitecture in FNdKO mice exposed substantial reductions in trabecular bone parameters and bone mineral density. The mice also showed elevated bone marrow adiposity. Analysis of chondrogenesis in FNdKO mice demonstrated changes in the resting, proliferating and hypertrophic growth plate zones, consistent alterations in chondrogenic markers such as collagen type II and X, decreased apoptosis of hypertrophic chondrocytes, and downregulation of bone formation markers. Transforming growth factor-β1 and downstream phospho-AKT levels were significantly lower in the FNdKO than in the control mice, revealing a crucial FN-mediated regulatory pathway in chondrogenesis and bone formation.In conclusion, the data demonstrate that FN is essential for chondrogenesis and bone development. Even though cFN and pFN act in different regions of the bone, both FN isoforms are required for the regulation of chondrogenesis, cartilage maturation, trabecular bone formation, and overall skeletal growth.

Apolipoprotein E is a novel marker for chondrocytes in the growth plate resting zone.

The resting zone (RZ) in mammalian growth plates is critical for maintaining and regulating chondrocyte turnover during longitudinal bone growth as a control tower and stem cell reservoir. Although recent lineage tracing studies have identified several markers for stem cells in the RZ, these markers only partially label chondrocytes in the RZ, suggesting that the resting chondrocytes (RCs) are a heterogeneous population with different types of stem cells. Since a comprehensive marker for RCs is still lacking, the RZ is generally determined based on ambiguous histological criteria, such as small and round chondrocytes without columnar formation, which may lead to inconsistencies among researchers. Therefore, in this study, we used single-cell RNA sequencing (scRNAseq) of growth plate chondrocytes followed by validation by fluorescence in situ hybridization (FISH) to precisely annotate cell clusters in scRNAseq and search for a marker of RCs. The scRNAseq analysis revealed that apolipoprotein E (Apoe) was the top-hit gene, which was ubiquitously expressed in the RC cluster. FISH confirmed that Apoe was exclusively localized to the histologically defined RZ. In newly generated Apoe-mCherry knock-in mice, we further confirmed that mCherry expression mirrored the distribution of Apoe-expressing chondrocytes in the RZ particularly after the formation of the secondary ossification center. These mCherry+ RCs were slow cycling in vivo and exhibited stem cell properties both in vitro and in vivo. Moreover, APOE was detected in human growth plate RCs. These findings suggest that Apoe is a novel pan-RC marker in both mouse and human growth plates.

Imaging Assessment of the Pediatric Elbow: Developmental Variants and Common Pathologies.

The pediatric elbow is a complex joint that undergoes rapid growth and development. The normal anatomy of the elbow varies depending on the age of the patient, which can be challenging for imaging interpretation. This article reviews developmental variants and common pathologies of the pediatric elbow, with a focus on their radiologic features. Normal anatomy and development of the pediatric elbow are discussed, including the six ossification centers and elbow alignment. Congenital anomalies such as longitudinal deficiencies of the upper extremity are reviewed. Some common injuries that affect the elbow, such as supracondylar fracture, lateral condyle fracture, medial epicondyle avulsion, and radial head dislocation are also described.

Pediatric Wrist.

Pediatric wrist injuries pose unique diagnostic challenges due to distinct bone characteristics in children and their diverse injury patterns. The dynamic development of the wrist, marked by changes in bone age and emerging ossification centers, is crucial to evaluate growth and identify potential pathologies. The skeletal composition, rich in cartilage, renders bones relatively weaker yet more elastic, impacting their susceptibility to fracture. Forearm fractures display diverse patterns influenced by torsional forces. Scaphoid fractures, less common in children, differ from those in adults. Conditions like Madelung's deformity and ulnar variance are more common wrist disorders in the pediatric population. In addition, the scarcity and nonspecificity of symptoms in those with tendon injuries and triangular fibrocartilage complex lesions can be diagnostically challenging. This article reviews pediatric wrist injuries, emphasizing ossification patterns, common fracture types, and developmental variants. Grasping these complexities in pediatric wrist development and associated pathologies is essential for precise diagnosis and treatment.

Imaging of Disease and Normal Variant Patterns in Pediatric Hips.

The pediatric hip undergoes significant changes from infancy through adolescence. Proper maturation is crucial for the development of a stable and functional hip joint. Imaging interpretation of the pediatric hip requires distinguishing normal variants and maturation patterns from pathology. We review femoral ossification centers, variants, and conditions that affect the proximal femur, such as Legg-Calvé-Perthes disease; the acetabulum, such as developmental hip dysplasia; the acetabular labrum, such as femoroacetabular impingement; and synovial pathology in children through adolescence. Understanding the spectrum of hip conditions and using advanced imaging techniques are essential for the accurate diagnosis and effective management of pediatric hip disorders.

Pediatric Foot: Development, Variants, and Related Pathology.

Pediatric foot development throughout childhood and adolescence can present a diagnostic dilemma for radiologists because imaging appearances may be confused with pathology. Understanding pediatric foot development and anatomical variants, such as accessory ossification centers, is essential to interpret musculoskeletal imaging in children correctly, particularly because many of these variants are incidental but others can be symptomatic. We first briefly review foot embryology. After describing common accessory ossification centers of the foot, we explain the different patterns of foot maturation with attention to irregular ossification and bone marrow development. Common pediatric foot variants and pathology are described, such as tarsal coalitions and fifth metatarsal base fractures. We also discuss pediatric foot alignment and various childhood foot alignment deformities.

Osteochondrosis of the primary ossification center (Kohler's Disease) of the patella

Osteochondrosis of the primary ossification center (Kohler's Disease) of the patella

Typical Development of the Secondary Ossification Centers of the Acetabulum and Their Effects on Acetabular Coverage of the Femoral Head.

We investigated the normal development of the secondary ossification centers of the acetabulum, focusing on their location and the amount of acetabular coverage increased by them. We enrolled 132 patients who were 7 to 16 years of age and had no pelvic deformity but did have ≥1 os ischium, os ilium, and/or os pubis on abdominal or pelvic computed tomographic (CT) scans. The locations of the ossification centers were evaluated by adopting an orientation using 0° for the superior acetabulum, 90° for the anterior acetabulum, 180° for the inferior acetabulum, and 270° for the posterior acetabulum, on a reconstructed 3-dimensional (3D) CT image. The acetabular coverage increase by the os ischium, os ilium, or os pubis was defined as the difference in the posterior acetabular sector angle (ΔPASA), posterosuperior acetabular sector angle (ΔPSASA), superior acetabular sector angle (ΔSASA), anterosuperior acetabular sector angle (ΔASASA), or anterior acetabular sector angle (ΔAASA) measured with and without each secondary ossification center. Patients were grouped into 3 age ranges: late childhood, preadolescence, and early adolescence. The location of each ossification center and the increase in acetabular coverage were compared between these groups. In the late-childhood group, the median start-to-end positions in right hips were 269° to 316° for the os ischium, 345° to 356° for the os ilium, and 81° to 99° for the os pubis. These positions tended to be wider in the early-adolescence group at 252° to 328° for the os ischium (p < 0.001), 338° to 39° for the os ilium (p = 0.005), and 73° to 107° for the os pubis (p = 0.049) in right hips. In right hips in the late-childhood group, the median values were 8.1° for ΔPASA, 14.0° for ΔPSASA, 9.9° for ΔSASA, 11.1° for ΔASASA, and 3.9° for ΔAASA; and in the early-adolescence group, the median values in right hips were 10.7° for ΔPASA, 12.9° for ΔPSASA, 8.4° for ΔSASA, 7.4° for ΔASASA, and 5.6° for ΔAASA. Only the median ΔPASA was larger in the early-adolescence group than in the late-childhood group (p = 0.026). Similar results were observed in left hips. In early adolescence, the secondary ossification centers appeared at more extended areas along the acetabular rim, and the increase in acetabular coverage by the secondary ossification centers tended to be larger in the posterior area but not in the anterior or superior area. Diagnostic Level III. See Instructions for Authors for a complete description of levels of evidence.

FOXC1 and FOXC2 regulate growth plate chondrocyte maturation towards hypertrophy in the embryonic mouse limb skeleton.

The Forkhead box transcription factors FOXC1 and FOXC2 are expressed in condensing mesenchyme cells at the onset of endochondral ossification. We used the Prx1-cre mouse to ablate Foxc1 and Foxc2 in limb skeletal progenitor cells. Prx1-cre;Foxc1Δ/Δ;Foxc2Δ/Δ limbs were shorter than controls, with worsening phenotypes in distal structures. Cartilage formation and mineralization was severely disrupted in the paws. The radius and tibia were malformed, whereas the fibula and ulna remained unmineralized. Chondrocyte maturation was delayed, with fewer Indian hedgehog-expressing, prehypertrophic chondrocytes forming and a smaller hypertrophic chondrocyte zone. Later, progression out of chondrocyte hypertrophy was slowed, leading to an accumulation of COLX-expressing hypertrophic chondrocytes and formation of a smaller primary ossification center with fewer osteoblast progenitor cells populating this region. Targeting Foxc1 and Foxc2 in hypertrophic chondrocytes with Col10a1-cre also resulted in an expanded hypertrophic chondrocyte zone and smaller primary ossification center. Our findings suggest that FOXC1 and FOXC2 direct chondrocyte maturation towards hypertrophic chondrocyte formation. At later stages, FOXC1 and FOXC2 regulate function in hypertrophic chondrocyte remodeling to allow primary ossification center formation and osteoblast recruitment.

P-253 Development of embryos from aged mothers transferred to the uterus of young mice

Abstract Study question Is the development of embryos from advanced maternal age (AMA) mice, similar to those from young maternal age (YMA) mice when transferred to young uteri? Summary answer AMA mouse embryos are characterized by altered organ morphology, particularly of the brain, which may predispose to neurodevelopmental disorders. What is known already Human and animal studies have linked AMA pregnancy to elevated risk of neurodevelopmental disorders and behavioural alterations in offspring, primarily attributed to compromised uterine (maternal) conditions. In human, it was also suggested that postnatal care of older mother may influence offspring behaviour. Animal studies revealed that behavioural alterations of AMA offspring are programmed during pregnancy and so they persist in litters fostered by young mouse. It remains unclear whether an embryo of older mother will undergo normal development after embryo transfer to younger female’s uterus. Study design, size, duration Embryos for Advanced Maternal Age, AMA (n = 105) and control Young Maternal Age, YMA (n = 44) groups were collected from thirty females 8-14 months old and six females 3-5 months old, respectively and transferred to pseudo-pregnant mice for further development. At embryonic day 16 (E16) foetuses were collected, fixed for histology and evaluated using Theiler and Kaufman’s classification of mouse development, where E16 corresponds to Theiler stage 24 (TS24) Participants/materials, setting, methods CBA x C57BL6 hybrid mice were used throughout the experiment. At E16, embryo survival rate (number of foetuses collected / number of transferred embryos) was scored for both groups. Foetuses were assessed for visible abnormalities and underwent histological evaluation of organs accordingly to TS 24. Parameters such as size/diameter, signs of vacuolization, cell density and layers of cerebral cortex in brain of foetuses were analysed. Values were considered different when p &amp;lt; 0.05, Mean±SD; Mann-Whitney test. Main results and the role of chance At E16, the survival of AMA embryos did not differ significantly from YMA control (39/105; 37% vs. 20/44; 45%, respectively) however, 5% of AMA foetuses (2/39) displayed oedema or cerebral haemorrhage. Histological analysis revealed alterations in the head/brain of AMA in comparison to YMA foetuses, such as smaller head size (head area in mid-sagittal section: 25.25±4.87 vs. 31.39±3.79 mm2, p &amp;lt; 0.05), thinner cerebral cortex (435.78±12.95 vs. 515.22±17.78µm, p &amp;lt; 0.005), reduced ratio of cortical plate (CP) to cerebral cortex thickness (0.20±0.03 vs. 0.26±0.009, p &amp;lt; 0.05) and an increased ratio of ventricular zone (VZ) to cerebral cortex thickness (0.29±0.002 vs. 0.24±0.002, p &amp;lt; 0.05). Moreover, decreased cellular density in CP layer (10378.86 vs. 12066.40 cells/mm2, P &amp;lt; 0.05) was noted. AMA brains were characterised by high degree of cellular vacuolisation, particularly in medulla oblongata, suggestive for spongiosis (1550.27 vs. 571.92 vacuoles/mm2, p &amp;lt; 0.05). These brain alterations may cause neurodevelopmental disorders in offspring. AMA foetuses had: smaller bronchial diameter (31.52 vs. 36.62µm, p &amp;lt; 0.05), no ossification centres and reduced skin wrinkling, corresponding to TS23 (that is previous developmental stage). These observations may suggest a delayed development of AMA foetuses. Limitations, reasons for caution The results of controlled animal experiments play a key role in developing conceptual models and formulating hypothesis about target species. However, extrapolation of these results to humans requires caution because species-specific developmental features may differ. Wider implications of the findings Our study demonstrates that advanced maternal age (AMA) impacts oocyte/embryo programming in mice. Morphological changes in AMA-conceived foetuses suggest potential risks for neurodevelopmental and other health issues in offspring, even when carried in the uterus of a young female, emphasizing the far-reaching effects of AMA on progeny health. Trial registration number not applicable

Radiological Assessment of Hand and Wrist Bones in Malnourished Children

Abstract Background Protein energy malnutrition (PEM) is still an important health problem that is responsible for childhood morbidity and mortality leading to permanent impairment of physical and mental growth. Objective This study is designed to assess the appearance of ossification centers of lower end of radius and ulna, carpals, metacarpals and phalanges in normal children and malnourished children then comparing between them, to detect early changes in bone growth in malnourished children. Subjects and Methods This case-control cross-sectional study included 45 malnourished children and 45 age and sex matched controls whose ages ranged between 6 months and 5 years. The patients were recruited from Pediatric Nutrition Clinic of Ain Shams University. All children (malnourished and control) were examined for anthropometric parameters (weight, height, body mass index (BMI) and mid upper arm circumference (MUAC)), investigated for serum calcium, phosphorus, alkaline phosphates and had a plain X-ray done on left hand and wrist (posteroanterior (P-A) view), the X-ray was analyzed by using A Digital Atlas of Skeletal Maturity. Results The plain X-ray hand and wrist showed delayed appearance of ossification centers in malnourished infants and children (97.8% had delayed bone age) when compared to controls (25.5% had delayed bone age) which was statistically highly significant with P-value &amp;lt; 0.01. Signs of vitamin D deficiency rickets together with serum calcium, phosphorus and alkaline phosphatase were statistically significantly lower in the cases when compared to the controls with P-value &amp;lt; 0.01. The statistical measurements of mean ± SD regarding weight of case group and control group was (9.05 ± 2.48, 13.54 ± 3.81) with P-value &amp;lt; 0.01 which is highly significant, regarding length or height was (79.51 ± 12.78, 88.94 ± 15.02) with P-value &amp;lt; 0.01 which is highly significant, regarding MUAC was (11.38 ± 1.91, 16.02 ± 1.26) with P-value &amp;lt; 0.01 which is highly significant. Conclusion X-ray hand and wrist for malnourished infants and children showed delayed appearance of ossification centers when compared to matched controls.