Intraskeletal bone growth patterns in the North Island Brown Kiwi (Apteryx mantelli): Growth mark discrepancy and implications for extinct taxa

Osteohistology, the study of bone microstructure, provides an important avenue for assessing extinct and extant vertebrate growth and life history. Cortical vascularity and collagen fibre organization are direct reflections of growth rate, while bone growth marks are indicative of absolute age. However, each skeletal element has its own ontogenetic trajectory and microstructure of certain bones may not be a true representation of whole body growth. Extensive comparative study of modern taxa is required to resolve intraskeletal discrepancies among age, vascularity and tissue organization in extinct vertebrates. Despite their comparative utility, studies of bone microstructure in modern taxa are severely lacking. Here, we add to a growing comparative osteohistological database by describing (1) bone tissue organization, (2) growth mark count, (3) sexually dimorphic bone (e.g. medullary bone) and (4) secondary cortical reconstruction in the bone microstructure of a 14-year-old male and 5-year-old female North Island Brown Kiwi (Apteryx mantelli). Transverse and longitudinal histological ground sections were processed and described for femora, tibiotarsi, tarsometatarsi, humeri, ulnae and radii in both kiwis. Cortical bone can generally be described as parallel-fibered tissue, interrupted by cyclical growth marks, with vascular canals oriented longitudinally within primary and secondary osteons. Tissue morphologically resembling medullary bone is present in the hindlimbs of the female, and coarse compacted cancellous bone (CCCB) is found sporadically in the male and female hindlimbs. Lines of arrested growth (LAGs) are present in all hindlimb bones of both kiwi, but remodelling has obliterated all LAGs in the male ulnae and radii. LAG count varies intraskeletally, but large weight bearing elements such as femora and tibiotarsi have less remodelling and, thus, higher number of LAGs. LAG count did not match absolute age in any skeletal element; a maximum of seven LAGs are present in the male kiwi and a maximum of seven LAGs in the female kiwi. The tissue organization within the forelimbs and hindlimbs is reflective of the protracted growth strategy of the North Island Brown Kiwi and congruent with previous studies of the kiwi. LAGs were highly variable throughout the skeleton of the kiwi and a decoupling of age and LAG deposition is apparent from the male kiwi samples. Excess LAGs in the 5-year-old female kiwi may be a product of hatching, egg laying or captivity. Regardless, LAG count variation in the kiwi stresses the importance of intraskeletal sampling when assessing growth patterns of extinct taxa. An extensive ontogenetic sampling of kiwi is necessary for future investigations of bone growth patterns, CCCB formation, medullary bone and LAG deposition and obliteration in these elusive birds.     

Bone remodeling in the longest living rodent,the naked mole-rat: Interelement variation and the effects of reproduction

The pattern of bone remodeling of one of the most peculiar mammals in the world, the naked mole-rat (NMR), was assessed. NMRs are known for their long lifespans among rodents and for having low metabolic rates. We assessed long-term in vivo bone labeling of subordinate individuals, as well as the patterns of bone resorption and bone remodeling in a large sample including reproductive and non-reproductive individuals (n = 70). Over 268 undecalcified thin cross-sections from the midshaft of humerus, ulna, femur and tibia were analyzed with confocal fluorescence and polarized light microscopy. Fluorochrome analysis revealed low osteogenesis, scarce bone resorption and infrequent formation of secondary osteons (Haversian systems) (i.e., slow bone turnover), thus most likely reflecting the low metabolic rates of this species. Secondary osteons occurred regardless of reproductive status. However, considerable differences in the degree of bone remodeling were found between breeders and non-breeders. Pre-reproductive stages (subordinates) exhibited quite stable skeletal homeostasis and bone structure, although the attainment of sexual maturity and beginning of reproductive cycles in female breeders triggered a series of anabolic and catabolic processes that up-regulate bone turnover, most likely associated with the increased metabolic rates of reproduction. Furthermore, bone remodeling was more frequently found in stylopodial elements compared to zeugopodial elements. Despite the limited bone remodeling observed in NMRs, the variation in the pattern of skeletal homeostasis (interelement variation) reported here represents an important aspect to understand the skeletal dynamics of a small mammal with low metabolic rates. Given the relevance of the remodeling process among mammals, this study also permitted the comparison of such process with the well-documented histomorphology of extinct therapsids (i.e., mammalian precursors), thus evidencing that bone remodeling and its endocortical compartmentalization represent ancestral features among the lineage that gave rise to mammals. It is concluded that other factors associated with development (and not uniquely related to biomechanical loading) can also have an important role in the development of bone remodeling.     

Morphological profile of atypical femoral fractures: age‐related changes to the cross‐sectional geometry of the diaphysis

The use of bisphosphonates for osteoporosis patients has markedly decreased the incidence of femoral neck or trochanteric fractures. However, anti‐osteoporosis drugs have been reported to increase the incidence of atypical femoral fractures, which involve stress fractures in the subtrochanteric region or the proximal diaphysis. In this study, the morphological characteristics of the cortical bone in human femoral diaphysis samples were analyzed from individuals who lived before bisphosphonate drugs were available in Japan. A total of 90 right femoral bones were arbitrarily selected (46 males and 44 females) from modern Japanese skeletal specimens. Full‐length images of these femurs were acquired using a computed tomography scanner. An image processing method for binarization was used to calculate the threshold values of individual bones for determining their contours. The range between the lower end of the lesser trochanter and the adductor tubercle of each femur was divided at regular intervals to obtain 10 planes. The mean value of cortical bone thickness, periosteal border length, and the cortical cross‐sectional area was evaluated for all planes. Moreover, the ratio of the area of the cortical bone to the total area of cross‐section at the mid‐diaphysis was calculated. A comparison between males and females demonstrated that most females had lower cortical bone area ratios at the mid‐diaphysis. The femoral outer shape did not differ markedly according to age or sex; however, substantial individual differences were observed in the shape of the inner surface of the cortical bone. The cortical bone thickness and the cross‐sectional area decreased with age in the femoral diaphysis; furthermore, in females, the decrease was higher for the former than for the latter. This may be due to a compensatory increase in the circumference of the femoral diaphysis. In addition, in about half of the subjects there was a discrepancy between the region with maximal value of the cortical bone thickness and that of the total cross‐sectional area. Biological responses to mechanical stresses to the femoral diaphysis are thought not to be uniform. Bisphosphonates inhibit bone resorption and may promote non‐physiological bone remodeling. Thus, a nonhomogeneous decrease in cortical thickness may be related to the fracture occurrence in the femoral diaphysis in some cases. Thus, long‐term administration of bisphosphonates in patients with morphological vulnerability in the femoral cortical bones may increase the occurrence of atypical femoral fractures.     

Functional bone histology of zebrafish reveals two types of endochondral ossification,different types of osteoblast clusters and a new bone type

The zebrafish is as an important vertebrate animal model system for studying developmental processes, gene functions and signalling pathways. It is also used as a model system for the understanding of human developmental diseases including those related to the skeleton. However, surprisingly little is known about normal zebrafish skeletogenesis and osteogenesis. As in most vertebrates, it is commonly known that the bones of adult zebrafish are cellular unlike that of some other teleosts. After careful histological analyses of each zebrafish adult bone, we identified several acellular bones, with no entrapped osteocytes in addition to several cellular bones. We show that both cellular and acellular bones can even occur within the same skeletal element and transitions between these two cell types can be found. Furthermore, we describe two types of osteoblast clusters during skeletogenesis and two different types of endochondral ossification. The epiphyseal plate, for example, lacks a zone of calcification and a degradation zone with osteoblasts. A new bone type that we term tubular bone was also identified. This bone is completely filled with adipose tissue, unlike spongy bones. This study provides important insight on how osteogenesis takes place in zebrafish, and especially on the transition from cellular to acellular bones. Overall, this study leads to a deeper understanding of the functional histological composition of adult zebrafish bones.     

去睾丸大鼠骨形态计量学的研究

目的：探讨4月龄雄性大鼠去睾丸后骨量的变化，重点比较松质骨和皮质骨骨代谢的变化。方法：20只4月龄SD雄性大鼠，随机分为假手术组（A组）和去睾丸组（B组），同等条件下饲养90d后，取腰椎和胫骨中段行不脱钙制片进行骨形态计量学观察。结果：去睾丸组与假手术组比较，前者腰椎松质骨吸收增加、骨形成下降（P〈0．05），出现明显骨质疏松；皮质骨外膜的骨形成动态参数如矿化沉积率、骨形成率下降（P〈0．05），内膜骨形成和吸收均有增加趋势。结论：去睾丸90d能使大鼠松质骨出现骨质疏松，皮质骨丢失不如松质骨显著。松质骨对去睾丸敏感性比皮质骨高。     

Development of Cortical Bone Geometry in the Human Femoral and Tibial Diaphysis

Ontogenetic growth processes in human long bones are key elements, determining the variability of adult bone structure. This study seeks to identify and describe the interaction between ontogenetic growth periods and changes in femoral and tibial diaphyseal shape. Femora and tibiae (n = 46) ranging developmentally from neonate to skeletally mature were obtained from the Norris Farms No. 36 archeological skeletal series. High‐resolution X‐ray computed tomography scans were collected. Whole‐diaphysis cortical bone drift patterns and relative bone envelope modeling activity across ages were assessed in five cross‐sections per bone (total bone length: 20%, 35%, 50%, 65%, and 80%) by measuring the distance from the section centroid to the endosteal and periosteal margins in eight sectors using ImageJ. Pearson correlations were performed to document and interpret the relationship between the cross‐sectional shape (I_max/I_min), total subperiosteal area, cortical area, and medullary cavity area for each slice location and age for both the femur and the tibia. Differences in cross‐sectional shape between age groups at each cross‐sectional position were assessed using nonparametric Mann‐Whitney U tests. The data reveal that the femoral and tibial midshaft shape are relatively conserved throughout growth; yet, conversely, the proximal and distal femoral diaphysis and proximal tibial diaphysis appear more sensitive to developmentally induced changes in mechanical loading. Two time periods of accelerated change are identified: early childhood and prepuberty/adolescence. Anat Rec, 296:774–787, 2013. © 2013 Wiley Periodicals, Inc.     

Rapid improvements in cortical bone dynamics and structure after lactation in established breeder rats

The mineral requirements for milk production during lactation usually result in a decrease in maternal skeletal mass during this period. The purpose of this study was to characterize changes in cortical bone formation, resorption, and structure after lactation in established breeder rats. Rats were taken at the end of the second pregnancy, second lactation, and at two, four, and six weeks after the pups were weaned. Age-related, nulliparous groups were included for comparison. Cortical bone structure and bone formation and resorption were measured at the tibiofibular junction using histomorphometric methods. As expected, there were decreases in cortical bone area, width, and minimum cortical thickness with an increase in marrow cavity area during lactation. Bone formation rates were essentially zero on the periosteal and endocortical surfaces at the end of lactation, while eroded (resorption) endocortical surface was greatly increased compared with the end of pregnancy. At or immediately after weaning, there was a rapid reversal of resorption to formation surface on the endocortical envelope similar to the events of true bone remodeling. Likewise, there was a commencement of bone formation on the periosteal surface. The volume- and surface-referent bone rates measured on the endocortical surface were substantially elevated over those measured at the end of the second pregnancy or in the nulliparous animals. Peak bone formation rates were observed on both the endocortical and periosteal surfaces by four weeks after weaning. These results show that the postlactation period is profoundly anabolic for cortical bone in the established breeder rat. The rapid and dramatic increases in bone formation likely serve to restore bone lost during lactation and to prepare the maternal skeleton for the next reproductive cycle.     

Microanatomical Record of Cortical Bone Remodeling and High Vascularity in a Fossil Giant Rat Midshaft Femur

Rat cortical bone does not typically undergo secondary (Haversian) remodeling. Haversian organization of rat bone has been mainly observed in experimental settings following biomechanical or dietary manipulation. Here, we report an observation of cortical secondary osteons within a histological femur cross-section from an extinct (late Quaternary) form of Timorese giant rat (Murinae gen. et sp. indet). The medio-lateral midshaft diameter of its femur, used as a measure of bone size, is 6.15 mm and indicates a heavier than normal skeletal frame. We compare this sample to bone histology in a small rat's midshaft femur of 2.33 mm diameter. A complete lack of Haversian bone remodeling characteristics is noted for the smaller sample, which is dominated by radial vascular canals. The giant rat shows clear secondary osteons and diffuse vascularity mainly composed of tightly packed longitudinal canals across its cortex. It appears that rat cortical bone can undergo bone remodeling, and is organized in a highly vascularized manner, in insular giant cases. Our findings from Timor align with results reported in experimental rat model skeletal biology literature and other insular fossil rat material. Where macroanatomical examination is limited, histological observations on fossil rat limb bones have the potential to aid reconstructions of life history and skeletal growth aspects in these rodents. Anat Rec, 302:1934–1940, 2019. © 2019 American Association for Anatomy     

Effect of hypokinesia on structure and mineral content of the long bones in puppies

The effect of hypokinesia for 2 months on the structure and mineral content of the cortical layer of the humerus, femur, and tibia was investigated in experiments on 6 puppies aged 2 months. The mineral content of the microstructures of the subperiosteal, subendosteal, and intermediate zones of the cortical layer of the long bones of the puppies, investigated by contact microroentgenography, increased under the influence of hypokinesia as a result of predominance of more mature structures resulting from delayed osteogenesis. An increase in the mean diameter of the Haversian canals was found as a result of increased osteoclastic absorption of bone tissue. Resorption also was intensified in the subperiosteal zone, as the result of which this zone and the whole cortical layer became thinner.     

Biomechanical properties of anuran long bones: correlations with locomotor modes and habitat use

Long bones are subjected to mechanical loads during locomotion that will influence their biomechanical properties through a feedback mechanism (the bone mechanostat). This mechanism adapts the spatial distribution of the mineralized tissue to resist compression, bending and torsion. Among vertebrates, anurans represent an excellent group to study long bone properties because they vary widely in locomotor modes and habitat use, which enforce different skeletal loadings. In this study, we hypothesized that (a) the cortical bone mass, density and design of anuran femur and tibiofibula would reflect the mechanical influences of the different locomotor modes and habitat use, and (b) the relationships between the architectural efficiency of cortical design (cross-sectional moments of inertia) and the intrinsic stiffness of cortical tissue [cortical mineral density; the 'distribution/quality' (d/q) relationship] would describe some inter-specific differences in the efficiency of the bone mechanostat to improve bone design under different mechanical loads. To test this hypothesis, we determined tomographic (peripheral quantitative computed tomography) indicators of bone mass, mineralization, and design along the femur and tibiofibula of four anuran species with different modes of locomotion and use of habitat. We found inter-specific differences in all measures between the distal and proximal ends and mid-diaphysis of the bones. In general, terrestrial-hopper species had the highest values. Arboreal-walker species had the lowest values for all variables except for cortical bone mineral density, which was lowest in aquatic-swimmer species. The d/q relationships showed similar responses of bone modeling as a function of cortical stiffness for aquatic and arboreal species, whereas terrestrial-hoppers had higher values for moments of inertia regardless of the tissue compliance to be deformed. These results provide new evidence regarding the significant role of movement and habitat use in addition to the biomechanical properties of long bones within a morpho-functional and comparative context in anuran species.     

The repair of fractured membrane bones in the newly hatched chick

Repair of a fractured membrane bone, the quadratojugal (QJ), has been studied in the newly hatched chick. Complete open fractures never united by bony fusion, even in birds maintained for six months post-fracture. Extraperiosteal connective tissue invaded the fracture gap and formed thick fibrous bundles which stabilised the fracture. Cartilage of two types formed on these bones. One was derived from periosteal cells and the other from osteoblasts or osteocytes. Considerably more cartilage formed in bones partially fractured than in those completely fractured. The “periosteal” cartilage did not form if the periosteum was removed at the time the bone was fractured. This was because, although the fibrous layer of the periosteum regenerated, the cambial layer did not. Metaplastic cartilage did form in the absence of the periosteum. Isolating fractured bones within polyethylene or glass tubes prevented accumulation of a blastema between the bony fragments. Cartilage did not form in side the tubes but did form where the ends of the tubes abutted onto the bones. Large defects in the bones (4 mm gaps, 4 mm of bone in the place of the QJ) healed via fibrous union with minimal osteogenesis and no chondrogenesis. Severing M. depressor mandibulae at the time the bone was fractured inhibited chondrogenesis, favoured osteogenesis and resulted in development of a pseudarthrosis. The potential for differentiation of the cells of the QJ and the role of adjacent tissues as they related to repair of the fracture was discussed, and the ability of cells from membrane bones to become chondrogenic emphasized.     

Effect of Y2 Receptor Deletion on Whole Bone Structural Behavior in Mice

Recent evidence has shown that mice deficient in the NPY Y2 receptor have an increase in trabecular bone volume as well as cortical bone mass due to increased osteoblast activity. However, the mechanical phenotype of Y2 ?/? bone has not yet been assessed. Thus, the aim of the present study was to examine the effect of Y2 deletion on murine cortical bone structural behavior, as well as to assess the material and geometric contributions to that behavior. The results of this study indicate that Y2 ?/? mice on a 129 SV × Balb/c background strain are smaller in body mass and have smaller bones than wild‐type controls. As expected based on smaller bone cross‐sectional properties, cortical bone structural strength was lower in ?/? animals. Surprisingly, the structural stiffness of ?/? bones was comparable to that of +/+ bones despite their smaller cross‐sectional geometry. Comparable structural stiffness appeared to be achieved by means of an elevated effective shear modulus, which was associated with a small, but statistically significant, higher ash content in Y2 ?/? bones. These data represent the first comprehensive characterization of the effect of Y2 deletion on cortical bone structural and material behavior to date. Anat Rec, 291:14–18, 2007. © 2007 Wiley‐Liss, Inc.     

Long bone histology of the subterranean rodent Bathyergus suillus (Bathyergidae): ontogenetic pattern of cortical bone thickening

Patterns of bone development in mammals are best known from terrestrial and cursorial groups, but there is a considerable gap in our understanding of how specializations for life underground affect bone growth and development. Likewise, studies of bone microstructure in wild populations are still scarce, and they often include few individuals and tend to be focused on adults. For these reasons, the processes generating bone microstructural variation at intra‐ and interspecific levels are not fully understood. This study comprehensively examines the bone microstructure of an extant population of Cape dune molerats, Bathyergus suillus (Bathyergidae), the largest subterranean mammal endemic to the Western Cape of South Africa. The aim of this study is to investigate the postnatal bone growth of B. suillus using undecalcified histological sections (n = 197) of the femur, humerus, tibia‐fibula, ulna and radius, including males and females belonging to different ontogenetic and reproductive stages (n = 42). Qualitative histological features demonstrate a wide histodiversity with thickening of the cortex mainly resulting from endosteal and periosteal bone depositions, whilst there is scarce endosteal resorption and remodeling throughout ontogeny. This imbalanced bone modeling allows the tissues deposited during ontogeny to remain relatively intact, thus preserving an excellent record of growth. The distribution of the different bone tissues observed in the cortex depends on ontogenetic status, anatomical features (e.g. muscle attachment structures) and location on the bone (e.g. anterior or lateral). The type of bone microstructure and modeling is discussed in relation to digging behavior, reproduction and physiology of this species. This study is the first histological assessment describing the process of cortical thickening in long bones of a fossorial mammal.     

脱细胞松质骨复合骨形态发生蛋白2异位成骨的实验研究

目的制备脱细胞松质骨作为骨形态发生蛋白2（BMP-2）载体。观察其异位成骨状况。方法取猪椎体松质骨,做脱细胞、脱脂处理。以兔为实验动物,将BMP-2与脱细胞松质骨复合,植入皮下组织．复合兔自体新鲜红骨髓及单纯脱细胞骨植入作为对照。另将脱细胞松质骨小柱复合BMP-2植入兔听泡内．以不含BMP-2脱细胞松质骨作为对照。术后3个月取标本行病理学检查。结果术后兔健康状况良好．术后3个月材料形状无明显变化．周围组织有少量血管增生．无明显免疫排斥现象,实验组病理学检查发现有新骨形成．对照组中复合自体红骨髓材料中有少量新生骨．单纯材料组仅有纤维结缔组织充填。植入听泡的复合BMP-2脱细胞松质骨与听泡骨壁接触部位结合紧密．表面被再生黏膜覆盖．病理学检查发现有新骨形成,而对照耳仅有黏膜覆盖,无新骨形成。结论脱细胞松质骨复合／BMP-2具有诱导成骨能力．并可在听泡空腔特殊部位诱导成骨．是一种很好的异位诱导成骨方法。     

Is the cortical capillary renamed as the transcortical vessel in diaphyseal vascularity?

A recent article published in Nature Metabolism, “A network of trans-cortical capillaries as a mainstay for blood circulation in long bones,” explained the long bone vascularity. In the mouse model, the authors demonstrated hundreds of transcortical vessels (TCVs) commencing from the bone marrow and traversing the whole cortical thickness. They realized that TCVs were the same as bleeding vessels of periosteal bed observed in the human tibia and femoral epiphysis during surgery. TCVs expressed arterial or venous markers and were proposed to be the backbone of bone vascularity as 80% of arterial and 59% of venous blood distributed through them. This new evidence challenged the existence of the “cortical capillaries” stated in previous literature. We conducted a review of the existing literature to compare this model with those in earlier research. The bone vascularity model was explained by many researchers who did their work in animal models like pig, dog, rabbit, and mouse. The TCVs were identified in these animal model studies as cortical capillaries or vessels of cortical canals. Studies are scarce, showing the presence of TCVs in humans. The role of TCVs in human cortical vascularity remains ambiguous until the substantial evidence is collected in future studies.     

骨形态计量学观察睾酮对雄性去势大鼠皮质骨的影响

目的通过骨形态计量方法观察雄激素替代疗法对去睾丸大鼠皮质骨代谢的影响。方法30只4月龄SD雄性大鼠,随机分成基础对照组(A组、实验开始时处死),年龄对照组(B组)、去睾丸组(C组)和去睾丸加睾丸酮组(D组),B组和C组生理盐水5ml·kg-1·d-1,D组甲基睾丸酮片1.8mg·kg-1·d-1,灌胃90d。实验结束,处死全部大鼠,取胫骨中段进行不脱钙骨制片,用计算机全自动图象分析系统进行骨组织形态计量学分析。结果去睾丸后皮质骨静态参数如截面总面积、髓腔面积等无明显变化,动态参数骨外膜骨形成降低(P<0.05),内膜骨形成和吸收均有增加趋势。睾酮则使去睾丸大鼠皮质骨的静态参数有增加趋势,促使骨外膜形成增加,减少内膜骨吸收(P<0.05),对内膜骨形成影响不大。结论睾酮补充治疗短期内能对抗去睾丸引起的大鼠皮质骨内外膜的代谢变化,维持正常的皮质骨结构。     

Prenatal cortical hyperostosis with COL1A1 gene mutation

Infantile cortical hyperostosis (Caffey disease) is benign and self-limiting when it presents near or after birth but it is usually lethal when it presents earlier. We present the clinical, ultrasonic, radiographic, and pathologic findings in an instructive case of early onset prenatal cortical hyperostosis. The pregnancy of a 21-year-old woman was medically terminated at 30 weeks of gestation after a diagnosis of severe osteogenesis imperfecta. Prenatal ultrasounds showed short long bones. Postmortem radiographs showed hyperostosis in long bones, ribs and mandible. The affected skeleton showed marked bony sclerosis and ballooning of the diaphyses of the long bones with periosteal sclerosis. A complete autopsy showed characteristic histologic findings of infantile cortical hyperostosis in affected bones. A missense mutation (3040C --> T) in exon 41 the gene encoding the alpha 1 chain of type I collagen was found in fetus pulmonary tissue. Neither the severe form nor the mild form of prenatal cortical hyperostosis were thought to be related to collagen I mutations. Our study indicates that a heterozygous 3040C --> T mutation can also be found in lethal prenatal cortical hyperostosis.     

Embryonic craniofacial bone volume and bone mineral density in Fgfr2+/P253R and nonmutant mice

Background : Quantifying multiple phenotypic aspects of individual craniofacial bones across early osteogenesis illustrates differences in typical bone growth and maturation and provides a basis for understanding the localized and overall influence of mutations associated with disease. We quantify the typical pattern of bone growth and maturation during early craniofacial osteogenesis and determine how this pattern is modified in Fgfr2^+/P253R Apert syndrome mice. Results: Early differences in typical relative bone density increase are noted between intramembranous and endochondral bones, with endochondral bones normally maturing more quickly during the prenatal period. Several craniofacial bones, including the facial bones of Fgfr2^+/P253R mice, display lower volumes during the earliest days of osteogenesis and lower relative densities until the perinatal period relative to unaffected littermates. Conclusions: Estimates of bone volume and linear measures describing morphology do not necessarily covary, highlighting the value of quantifying multiple facets of gross osteological phenotypes when exploring the influence of a disease causing mutation. Differences in mechanisms of osteogenesis likely underlie differences in intramembranous and endochondral relative density increase. The influence of the FGFR2 P253R mutation on bone volume changes across the prenatal period and again after birth, while its influence on relative bone density is more stable. Developmental Dynamics 243:541–551, 2014. © 2013 Wiley Periodicals, Inc.