Haversian Remodeling Research Articles

Wing phalanges of a ?thalassodromine pterosaur from the Aptian–Albian Antlers Formation of Texas, USA

Fragmentary pterosaurian wing phalanges from the Aptian-Albian Antlers Formation of north-central Texas, USA are described. Wing phalanges 1 and 2 have suboval shafts with thickened bone at the leading and trailing edges, whereas the third wing phalanx has a flattened airfoil-like cross-section with greatly thickened bone at the leading and trailing edges and a small subrectangular medullary cavity. The cortical bone exhibits an unusual coarse surface texture, and bone thin sections reveal extensive Haversian remodeling and endosteal lamellar bone. The morphology of the wing phalanges differs from that of dsungaripterids, pteranodontoids, and azhdarchids, so the specimen is not a North American record of the Dsungaripteridae as some have stated; whereas based on comparisons to a roughly contemporaneous indeterminate thalassodromine pterosaur specimen from the Romualdo Formation of Brazil, the specimen is tentatively interpreted as an indeterminate thalassodromine with an estimated wingspan of ∼3.5 m.

Prolonged cyclical loading induces Haversian remodeling in mandibles of growing rabbits.

Bone adaptation to mechanical loading happens predominantly via modeling and remodeling, but the latter is poorly understood. Haversian remodeling (cortical bone replacement resulting in secondary osteons) is thought to occur in regions of low strain as part of bone maintenance or high strain in response to microdamage. However, analyses of remodeling in primates have revealed an unappreciated association with the number of daily load cycles. We tested this relationship by raising 30 male domestic rabbits (Oryctolagus cuniculus) on disparate diets from weaning to adulthood (48 weeks), facilitating a naturalistic perspective on mandibular bone adaptation. A control group consumed only rabbit pellets and an 'overuse' group ate hay in addition to pellets. To process hay, which is tougher and stiffer, rabbits increase chewing investment and duration without increasing bite force (i.e. corpus mean peak strain is similar for the two foods). Corpus remodeling in overuse rabbits was ∼1.5 times that of controls, measured as osteon population density and percentage Haversian bone. In the same subjects, there was a significant increase in overuse corpus bone formation (ratio of cortical area to cranial length), consistent with previous reports on the same dietary manipulation and bone formation in rabbits. This is the first evidence that both modeling and remodeling are simultaneously driven by the number of load cycles, independent of strain magnitude. This novel finding provides unique data on the feeding apparatus, challenges traditional thought on Haversian remodeling, and highlights the need for experimental studies of skeletal adaptation that examine mechanical factors beyond strain magnitude.

Bone microstructure of bony-toothed birds (Odontopterygiformes) from the Eocene of Ikove, Ukraine: preliminary paleobiological implications

ABSTRACT Histological structure of long bones from bony-toothed birds (Odontopterygiformes) from Early Lutetian locality Ikove (Ukraine) is examined for the first time. Six limb bones from Pelagornithidae cf. Dasornis sp. and Lutetodontopteryx tethyensis are described. The bones, although usually being fragmented, show good preservation at the microscopic level. Their microstructure (e.g. orientation and diameter of primary osteons) significantly varies, but the basic pattern and, therefore, the way of growth matches that of recent Neognathae (as well as a number of extinct taxa). In young birds, the periosteum quickly deposited well-vascularised primary bone, which at the same time was resorbed from inside, then the animal reached adult size and fast bone growth gave way to much slower deposition of circumferential layers of lamellar or parallel-fibred tissue on the outer and inner bone surfaces. The bones also underwent Haversian remodelling of various extent (from nearly absent to nearly full), generally stronger in larger bones and in the inner compacta. Bone microstructure is consistent with high metabolic rates, expected in Odontopterygiformes, and implies that their growth rates were comparable to those of recent Neognathae.

Absence of secondary osteons in femora of aged rats: Implications of lifespan on Haversian remodeling in mammals.

Bone is a dynamic tissue capable of adapting to its loading environment, allowing the skeleton to remain structurally sound throughout life. One way adaptation occurs in mammals is via Haversian remodeling: the site-specific, coupled resorption and formation of cortical bone that results in secondary osteons. Remodeling occurs at a baseline rate in most mammals, but it also occurs in relation to strain by repairing deleterious microdamage. Yet, not all animals with bony skeletons remodel. Among mammals, there is inconsistent or absent evidence for Haversian remodeling among monotremes, insectivores, chiropterans, cingulates, and rodents. Three possible explanations for this disparity are discussed: the capacity for Haversian remodeling, body size as a constraint, and age and lifespan as constraints. It is generally accepted, although not thoroughly documented, that rats (a common model used in bone research) do not typically exhibit Haversian remodeling. The present aim is to more specifically test the hypothesis that rats of advanced age do remodel intracortically because of the longer lifespan over which baseline remodeling could occur. Most published histological descriptions of rat bone only include young (3-6 months) rats. Excluding aged rats possibly overlooks a transition from modeling (i.e., bone growth) to Haversian remodeling as the primary mode of bone adaptation. Here, midshaft and distal femora (typical sites for remodeling in other mammals) of 24-month-old rats were examined for presence of secondary osteons. None were found, suggesting that Haversian remodeling does not occur in rats under normal physiological conditions at any age. A likely explanation is that modeling of cortical bone continues throughout most of the short rat lifespan, negating the stimulus for Haversian remodeling. Thorough sampling of key rodent taxa of varying body sizes and lifespans is key to elucidating the reasons why (i.e., body size, age/lifespan, phylogenetic factors) Haversian remodeling might not occur in all mammals.

Pachyosteosclerosis, rhamphotheca and enhanced sensory capabilities of the premaxillae of Hyperodapedon (Archosauromorpha, Rhynchosauria): implications for foraging at the sediment–water interface

AbstractThe external morphology and microanatomy of 32 partial and complete Hyperodapedon premaxillae was examined to assess their functional attributes. This revealed morphological correlates for innervation of Hyperodapedon premaxillae in the form of posteriorly opening enlarged neurovascular foramina associated with several grooves, and a prominent neurovascular sulcus. Scanning electron microscopy shows numerous small, circular foramina in clusters along the lateroventral surface towards the anterior tip and along the ventral edge, often in a preferred orientation. These are found associated with high rugosity along the elongated anterolateral depression, and were related to nutrient supply and/or part of the neurovascular system. Selected premaxillae show extremely high bone compactness indices (especially at the anterior end) suggesting specialized osteosclerotic conditions, and dense and compact bone microstructure with almost no clear transition between the outer compact cortex and inner core. With ontogeny, the premaxillae became lateromedially thickened by deposition of lamellar zonal bone, and highly vascularized and dense from intense Haversian remodelling, suggesting pachyosteosclerosis of the premaxillae. Other characteristic features include profuse open vascular channels or a frayed margin at the anteroventral tip, and dense bundles of long and wavy extrinsic fibres. These features, along with high bone compactness, decrease posteriorly towards the naris. It is proposed that the Hyperodapedon premaxillae were covered by keratinized epithelium or rhamphotheca at the anterior end, and had heightened sensory capabilities that aided foraging for mussels and other invertebrates in soft sediments under shallow water. Such enhanced sensory capability is reported for the first time in an early‐diverging archosauromorph.

Haversian Remodeling in Juvenile White‐Tailed Deer Takes Place Prominently in the Humerus Proximal Diaphysis

During the course of Haversian remodeling, existing cortical bone tissue is being resorbed by osteoclasts, forming cutting cones, which then are filled by osteoblasts, depositing osteoid in concentric lamellae and forming secondary osteons (Haversian systems). This continues process, is induced, at least in part, by accumulation of microcracks in the tissue. In large mammals, such as deer, this process is responsible for the change of fast-deposited juvenile primary plexiform bone, into mature secondary Haversian bone. The objective of this study was to determine if Haversian remodeling in juvenile white-tailed deer is more prominent in forelimb vs. hindlimb long bones (humerus and femur respectively), and if Haversian remodeling initiates earlier in the proximal vs. mid diaphysis of these bones. The first hypothesis is that since the forelimb supports more of the deer weight, it is subjected to larger stresses and strains and it will accumulate more microcracks, and thus the humerus will reveal earlier and more prominent Haversian remodeling compared to the femur. The second hypothesis is that due to expected larger bending moments in the mid-diaphysis region of both the humerus and femur, the mid-diaphysis region will accumulate more microcracks, and thus will demonstrate earlier and more prominent Haversian remodeling compared to the proximal diaphysis. To this end, light and electron microscopy was used to inspect multiple sections from humeri and femora harvested from seven juvenile white-tailed deer. Deer juvenile state was determined by the existence of an active growth plate. In addition, multiple bone samples from these regions were crushed, grounded and ashed to quantify their water and mineral content, as higher water content and lower mineral content are both indicative of newer and not fully mineralized bone tissue, which is the initial outcome of Haversian remodeling. The microscopy bone slides revealed that the humerus, and especially the proximal diaphysis region, demonstrated earlier and more prominent Haversian remodeling compared to the femur. These relatively qualitative results were further supported by significantly higher water content and lower mineral content values in the proximal and mid-diaphysis humerus compared to the femur, and in the proximal diaphysis compared to the mid-diaphysis in both bones (see Figure 1). These results support the prediction of more prominent and early Haversian remodeling in the forelimb compared to the hindlimb of white-tailed deer. Yet, contrary to the predictions of the second hypothesis, both the humerus and femur proximal diaphyses, and not mid-diaphyses, demonstrated earlier and more prominent Haversian remodeling. This is potentially due to the increase in bone turnover activity around the active proximal growth plate. In conclusion, this study reveals that Haversian remodeling starts earlier and is more prominent in the humerus compared to the femur and in the proximal diaphysis compared to the mid-diaphysis of juvenile white-tailed deer. These data can potentially help in approximating the age of white-tailed deer long bones, when more precise data is missing.

Bone remodeling and cyclical loading in maxillae of New Zealand white rabbits (Oryctolagus cuniculus).

Mammalian feeding behaviors are altered when mechanically challenging (e.g., tough, stiff) foods require large bite forces or prolonged mastication. Bony responses to high bite forces are well-documented for the mammalian skull, but osteogenesis due to cyclical loading, caused by repetitive chewing, is more poorly understood. Previous studies demonstrate that cyclical loading results in greater bone formation in the rabbit masticatory apparatus and in substantial Haversian remodeling in primate postcrania. Here we assess the relationship between cyclical loading and remodeling in the rabbit maxilla. Twenty male New Zealand white rabbits (Oryctolagus cuniculus) were raised on either an overuse or control diet (10 per group) for 48 weeks, beginning at weaning onset. The control group was raised on a diet of rabbit pellets (E=29 MPa, R=1031 J/m2 ), whereas the overuse group ate rabbit pellets and hay, which has high stiffness (E=3336 MPa) and toughness (R=2760 J/m2 ) properties. Hay requires greater chewing investment (475 chews/g) and longer chewing durations (568 s/g) than pellets (161 chews/g and 173 s/g), therefore causing cyclical loading of the jaws. Remodeling was measured as osteon population density (OPD), percent Haversian bone (%HAV), and osteon cross-sectional area (On.Ar). The only significant difference found was greater On.Ar in the alveolar region of the maxilla (p < 0.001) in the overuse group. The hypothesis that cyclical loading engenders Haversian remodeling in the developing maxilla is not supported. The continuation of modeling throughout the experimental duration may negate the need for remodeling as newly laid bone tends to be more compliant and resistant to crack propagation.

Humerus midshaft histology in a modern and fossil wombat

The common wombat (Vombatus ursinus) is equipped with a set of physiological and morphological adaptations suited to a fossorial lifestyle. These allow wombats to engage in efficient scratch-digging and maintaining a low basal metabolic rate while living underground. While bone microstructure has been described for several subterranean animals, wombat bone histology has received very little attention to date. Here, we present preliminary insights into bone histology in modern adult V. ursinus (Mt Fairy, New South Wales) and Pleistocene fossil Vombatus sp. (Bakers Swamp, New South Wales) midshaft humeri. The modern sample was well preserved, allowing us to identify varying bone tissue types (woven, parallel-fibred, lamellar). The sample showed vascularity composed of primary and secondary osteons, and simple longitudinal and radial vessels. We also observed evidence for Haversian remodelling (i.e. localised replacement of pre-existing bone) and coarse compact cancellous bone within the inner cortex of the diaphysis. The fossil histology was poorly preserved, but likely showed bone matrix organisation similar to the modern specimen. We use these preliminary data to discuss hypotheses for wombat forelimb biomechanical and physiological microscopic adaptation to a burrow environment. We encourage future intraskeletal examination of microstructure in wombat populations to better inform their ecological adaptations and behaviour in palaeontological contexts.

Bone Biomineral Properties Vary across Human Osteonal Bone.

The biomineralization of bone remains a puzzle. During Haversian remodeling in the dense human cortical bone, osteoclasts excavate a tunnel that is then filled in by osteoblasts with layers of bone of varying fibril orientations, resulting in a lamellar motif. Such bone represents an excellent possibility to increase our understanding of bone as a material as well as bone biomineralization by studying spatio/temporal variations in the biomineral across an osteon. To this end, fluorescence computed tomography and diffraction scattering computed tomography with sub-micrometer resolution is applied to obtain position resolved fluorescence spectra and diffraction patterns in a 3D volume. The microstructural properties of the apatite biomineral are not homogeneous but depend critically on the time point at which it was laid down. This indicates that the nature of bone biomineral is highly dependent on the microenvironment during bone formation and remodeling.

Histovariability in human clavicular cortical bone microstructure and its mechanical implications.

The human clavicle (i.e. collarbone) is an unusual long bone due to its signature S-shaped curve and variability in macrostructure observed between individuals. Because of the complex nature of how the upper limb moves, as well as due to its complex musculoskeletal arrangement, the biomechanics, in particular the mechanical loadings, of the clavicle are not fully understood. Given that bone remodeling can be influenced by bone stress, the histologic organization of Haversian bone offers a hypothesis of responses to force distributions experienced across a bone. Furthermore, circularly polarized light microscopy can be used to determine the orientation of collagen fibers, providing additional information on how bone matrix might organize to adapt to direction of external loads. We examined Haversian density and collagen fiber orientation, along with cross-sectional geometry, to test whether the clavicle midshaft shows unique adaptation to atypical load-bearing when compared with the sternal (medial) and acromial (lateral) shaft regions. Because fractures are most common at the midshaft, we predicted that the cortical bone structure would show both disparities in Haversian remodeling and nonrandomly oriented collagen fibers in the midshaft compared with the sternal and acromial regions. Human clavicles (n=16) were sampled via thin-sections at the sternal, middle, and acromial ends of the shaft, and paired sample t-tests were employed to evaluate within-individual differences in microstructural or geometric properties. We found that Haversian remodeling is slightly but significantly reduced in the middle of the bone. Analysis of collagen fiber orientation indicated nonrandom fiber orientations that are overbuilt for tensile loads or torsion but are poorly optimized for compressive loads throughout the clavicle. Geometric properties of percent bone area, polar second moment of area, and shape (Imax /Imin ) confirmed the conclusions drawn by existing research on clavicle macrostructure. Our results highlight that mediolateral shape changes might be accompanied by slight changes in Haversian density, but bone matrix organization is predominantly adapted to resisting tensile strains or torsion throughout and may be a major factor in the risk of fracture when experiencing atypical compression.

Haversian remodeling corresponds to load frequency but not strain magnitude in the macaque (Macaca fascicularis) skeleton.

One way bone adapts to its mechanical environment is by Haversian remodeling, a repair process in which existing bone is resorbed and replaced by new bone. Haversian remodeling forms interconnected, cylindrical structures called secondary osteons. The amount of remodeling that occurs is related to the nature of mechanical loading and accrual of microdamage, but it is uncertain whether habitual loads of high magnitude versus high frequency result in more remodeling. The answer to this question is important if remodeling is to be a tool for inferring loading environments, and thus behavior, in past populations. Here, secondary osteon population density (OPD), osteon cross-sectional area (On.Ar), and percent Haversian bone (%HAV) were compared among mid-diaphysis femora, tibia, fibulae, and mid-level ribs of five adult crab-eating macaques (Macaca fascicularis). Ribs experience relatively low strains but have a high daily loading frequency (~33 times per minute). Limb bones are loaded for fewer cycles per day, but the femur and tibia have high load magnitudes due to gravitational forces. Strain magnitudes in the fibula are a fraction of those in the femur and tibia. Analyses of variance demonstrated significant differences in OPD (P = 0.010) and On.Ar (P < 0.001) among the bones. Pairwise t-tests revealed greater OPD but lower On.Ar in the rib than all other bones. The high rib OPD suggests that Haversian remodeling is more responsive to load frequency than strain magnitude. The fact that osteons are smaller in ribs than any other bone may be an effect of remodeling in comparatively narrow cortices.

Rabbit as model for osteoporosis research.

Osteoporosis is a major public health problem affecting more than 200 million people worldwide. The use of different animal models, for the study of its pathophysiology and treatments, is important being actually the ovariectomized rat the most widely used; although this model has several problems due its small size, lack of true closure of epiphyseal plate and bone differences with humans. This review is aimed at summarizing the most common methods published for osteoporosis induction in rabbits as model for human disease with their advantages and disadvantages. The paper shows the advantages of the use of this specie compared with the rat. All the techniques seemed to achieve the osteoporotic condition, but the one which obtained the most consistent bone mineral reduction in less time was the combination of surgery and corticoid treatment. The conclusion of the review was that rabbits are promising as a model of osteoporosis research because of their size, haversian remodelling and closure of epiphyseal plate, which solve some of the problems of the rat model. There are different techniques in the literature used to achieve the osteoporotic condition with diverse results, but there is a lack of consensus as to the best one.

Role of porosity and matrix behavior on compressive fracture of Haversian bone using random spring network model.

Haversian remodeling is known to result in improved resistance to compressive fracture in healthy cortical bone. Here, we examine the individual roles of the mean porosity, structure of the network of pores and remodeled bone matrix properties in the fracture behavior of Haversian bone. The detailed structure of porosity network is obtained both pre- and post-testing of dry cubical bone samples using micro-Computed Tomography. Based on the periodicity in the features of porosity along tangential direction, we develop a two dimensional porosity-based random spring network model for Haversian bone. The model is shown to capture well the macroscopic response and reproduce the avalanche statistics similar to recently reported experiments on porcine bone. The predictions suggest that at the millimeter scale, the remodeled bone matrix of Haversian bone is less stiff but tougher than that of plexiform/primary bone.

Development of an in vivo bone fatigue damage model using axial compression of the rabbit forelimb

Many nontraumatic fractures seen clinically in patients with metabolic bone disorders or on antiresorptive treatment show an increased incidence of microdamage accumulation and impaired intracortical remodeling. However, the lack of basal remodeling and Haversian bone in rodents limits their translatability in studying bone damage repair mechanisms. The work presented here demonstrates the development of the forelimb loading model in rabbits, the smallest mammal with intracortical Haversian remodeling. The forelimbs of post-mortem female New Zealand white rabbits were loaded in axial end compression to determine their basic monotonic and fatigue properties. Following time zero characterization, stress fractures were created in vivo and animals were allowed to recover for a period of two to five weeks. The rabbit forelimb when loaded in axial compression demonstrates a consistent mid-diaphyseal fracture location characterized by a local mixed compression-bending loading environment. Forelimb apparent stiffness, when fatigue loaded, demonstrates a progressive increase until macrocrack formation, at which time apparent stiffness rapidly declines until failure. Stress fractures in the rabbit ulna display robust periosteal expansion and woven bone formation two weeks following fracture. Subsequent healing at five weeks post-fracture is marked by woven bone densification, resorption and intracortical remodeling along the stress fracture line. The rabbit forelimb fatigue model is a promising new platform by which bone׳s response to damage may be studied.

Structure and growth pattern of the bizarre hemispheric prominence on the rostrum of the fossil beaked whale Globicetus hiberus (Mammalia, Cetacea, Ziphiidae).

The rostrum of most ziphiids (beaked whales) displays bizarre swollen regions, accompanied with extreme hypermineralisation and an alteration of the collagenous mesh of the bone. The functional significance of this specialization remains obscure. With the voluminous and dense hemispheric excrescence protruding from the premaxillae, the recently described fossil ziphiid Globicetus hiberus is the most spectacular case. This study describes the histological structure and interprets the growth pattern of this unique feature. Histologically, the prominence in Globicetus is made up of an atypical fibro-lamellar complex displaying an irregular laminar organization and extreme compactness (osteosclerosis). Its development is suggested to have resulted from a protraction of periosteal accretion over the premaxillae, long after the end of somatic growth. Complex shifts in the geometry of this tissue are likely to have occurred during its accretion and no indication of Haversian remodeling could be found. X-ray diffraction and Raman spectroscopy indicate that the bone matrix in the premaxillary prominence of Globicetus closely resembles that of the rostrum of the extant beaked whale Mesoplodon densirostris: apatite crystals are of common size and strongly oriented, but the collagenous meshwork within bone matrix seems to be extremely sparse. These morphological and structural data are discussed in the light of functional interpretations proposed for the highly unusual and diverse ziphiid rostrum. J. Morphol. 277:1292-1308, 2016. © 2016 Wiley Periodicals, Inc.

Immobilization and long-term recovery results in large changes in bone structure and strength but no corresponding alterations of osteocyte lacunar properties

The ability of osteocytes to demineralize the perilacunar matrix, osteocytic osteolysis, and thereby participate directly in bone metabolism, is an aspect of osteocyte biology that has received increasing attention during the last couple of years. The aim of the present work was to investigate whether osteocyte lacunar properties change during immobilization and subsequent recovery. A rat cortical bone model with negligible Haversian remodeling effects was used, with temporary immobilization of one hindlimb induced by botulinum toxin. Several complementary techniques covering multiple length scales enabled correlation of osteocyte lacunar properties to changes observed on the organ and tissue level of femoral bone. Bone structural parameters measured by μCT and mechanical properties were compared to sub-micrometer resolution SR μCT data mapping an unprecedented number (1.85 million) of osteocyte lacunae.Immobilization induced a significant reduction in aBMD, bone volume, tissue volume, and load to fracture, as well as the muscle mass of rectus femoris. During the subsequent recovery period, the bone structural and mechanical properties were only partly regained in spite of a long-term (28weeks) study period. No significant changes in osteocyte lacunar volume, density, oblateness, stretch, or orientation were detected upon immobilization or subsequent recovery. In conclusion, the bone architecture and not osteocyte lacunar properties or bone material characteristics dominate the immobilization response as well as the subsequent recovery.

Development of an in vivo rabbit ulnar loading model

Ulnar and tibial cyclic compression in rats and mice have become the preferred animal models for investigating the effects of mechanical loading on bone modeling/remodeling. Unlike rodents, rabbits provide a larger bone volume and normally exhibit intracortical Haversian remodeling, which may be advantageous for investigating mechanobiology and pharmaceutical interventions in cortical bone. Therefore, the objective of this study was to develop and validate an in vivo rabbit ulnar loading model. Ulnar tissue strains during loading of intact forelimbs were characterized and calibrated to applied loads using strain gauge measurements and specimen-specific finite element models. Periosteal bone formation in response to varying strain levels was measured by dynamic histomorphometry at the location of maximum strain in the ulnar diaphysis. Ulnae loaded at 3000 microstrain did not exhibit periosteal bone formation greater than the contralateral controls. Ulnae loaded at 3500, 4000, and 4500 microstrain exhibited a dose-dependent increase in periosteal mineralizing surface (MS/BS) compared with contralateral controls during the second week of loading. Ulnae loaded at 4500 microstrain exhibited the most robust response with significantly increased MS/BS at multiple time points extending at least 2weeks after loading was ceased. Ulnae loaded at 5250 microstrain exhibited significant woven bone formation. Rabbits required greater strain levels to produce lamellar and woven bone on periosteal surfaces compared with rats and mice, perhaps due to lower basal levels of MS/BS. In summary, bone adaptation during rabbit ulnar loading was tightly controlled and may provide a translatable model for human bone biology in preclinical investigations of metabolic bone disease and pharmacological treatments.

Mammalian cortical bone in tension is non-Haversian.

Cortical bone, found in the central part of long bones like femur, is known to adapt to local mechanical stresses. This adaptation has been linked exclusively with Haversian remodelling involving bone resorption and formation of secondary osteons. Compared to primary/plexiform bone, the Haversian bone has lower stiffness, fatigue strength and fracture toughness, raising the question why nature prefers an adaptation that is detrimental to bone's primary function of bearing mechanical stresses. Here, we show that in the goat femur, Haversian remodelling occurs only at locations of high compressive stresses. At locations corresponding to high tensile stresses, we observe a microstructure that is non-Haversian. Compared with primary/plexiform bone, this microstructure's mineralisation is significantly higher with a distinctly different spatial pattern. Thus, the Haversian structure is an adaptation only to high compressive stresses rendering its inferior tensile properties irrelevant as the regions with high tensile stresses have a non-Haversian, apparently primary microstructure.

Calcified Cartilage Islands in Rat Cortical Bone

Rats display little to no haversian remodeling of cortical bone. This fact, combined with the endochondral formation of cortical bone, means that rat femoral cortical bone contains highly mineralized cartilage islands in a central band of mid-femoral cross sections. We demonstrate that these islands have a significantly higher degree of mineralization than the surrounding bone, using quantitative backscattered electron imaging. The cartilaginous nature of the islands was verified by immunostaining for collagen type II. Toluidine blue staining of longitudinal sections and three-dimensional synchrotron radiation X-ray tomographic microscopy confirmed that the islands are elongated along the femoral long axis. Nanoindentation revealed significantly higher values of both reduced modulus and hardness in the islands compared to the surrounding bone, reflecting a higher degree of mineralization. The calcified cartilage islands were distributed in a central zone of the bone, from the growth plates through the mid-femoral bone. The presence of these cartilage islands and their possible effect on mechanical properties could be an additional reason why haversian remodeling is observed in higher-order species.

Evolutionary implications of the divergent long bone histologies of Nothosaurus and Pistosaurus (Sauropterygia, Triassic)

BackgroundEosauropterygians consist of two major clades, the Nothosauroidea of the Tethysian Middle Triassic (e.g., Nothosaurus) and the Pistosauroidea. The Pistosauroidea include rare Triassic forms (Pistosauridae) and the Plesiosauria of the Jurassic and Cretaceous. Long bones of Nothosaurus and Pistosaurus from the Muschelkalk (Middle Triassic) of Germany and France and a femur of the Lower Jurassic Plesiosaurus dolichodeirus were studied histologically and microanatomically to understand the evolution of locomotory adaptations, patterns of growth and life history in these two lineages.ResultsWe found that the cortex of adult Nothosaurus long bones consists of lamellar zonal bone. Large Upper Muschelkalk humeri of large-bodied Nothosaurus mirabilis and N. giganteus differ from the small Lower Muschelkalk (Nothosaurus marchicus/N. winterswijkensis) humeri by a striking microanatomical specialization for aquatic tetrapods: the medullary cavity is much enlarged and the cortex is reduced to a few millimeters in thickness. Unexpectedly, the humeri of Pistosaurus consist of continuously deposited, radially vascularized fibrolamellar bone tissue like in the Plesiosaurus sample. Plesiosaurus shows intense Haversian remodeling, which has never been described in Triassic sauropterygians.ConclusionsThe generally lamellar zonal bone tissue of nothosaur long bones indicates a low growth rate and suggests a low basal metabolic rate. The large triangular cross section of large-bodied Nothosaurus from the Upper Muschelkalk with their large medullary region evolved to withstand high bending loads. Nothosaurus humerus morphology and microanatomy indicates the evolution of paraxial front limb propulsion in the Middle Triassic, well before its convergent evolution in the Plesiosauria in the latest Triassic. Fibrolamellar bone tissue, as found in Pistosaurus and Plesiosaurus, suggests a high growth rate and basal metabolic rate. The presence of fibrolamellar bone tissue in Pistosaurus suggests that these features had already evolved in the Pistosauroidea by the Middle Triassic, well before the plesiosaurs radiated. Together with a relatively large body size, a high basal metabolic rate probably was the key to the invasion of the Pistosauroidea of the pelagic habitat in the Middle Triassic and the success of the Plesiosauria in the Jurassic and Cretaceous.