Small Animal Bones Research Articles

To the Question of the Purpose of Some Types of Bone Products from Medieval Sites of the Gis-Urals and Trans-Urals: to the Formulation of the Problem

The article is devoted to the analysis of one category of bone products discovered during archaeological research at settlement complexes of the XII–XIV centuries of the Tobol and Irtysh Rivers Basin (Western Siberia, Trans-Urals). The artifacts are small bone cases with a hole for hanging, inside of them there was one bone tool with a sharpened working edge, analogies of which are found on medieval sites of the Permian Gis-Urals, attributed by researchers as needles / punctures located in needle cases. However, the morphological and structural characteristics of these artifacts, in our opinion, do not correspond to the functions attributed to them. The purpose of the study is to introduce into scientific circulation and attribution of the actual purpose of paired bone products found on medieval sites of the Tobolo–Irtysh region. Based on the purpose of the study, the following tasks were put forward: morphological description of products; search for analogies to objects on archaeological sites of adjacent territories; acquaintance with the history of the study of similar artifacts. To solve the tasks set in the work, traditional research methods are used for historical science: comparative-historical, comparative-typological, formal-stylistic, the method of analogies. The morphological analysis of the finds both from the Tobol-Irtysh region and from the sites of the Gis-Urals, coupled with the analysis of a complex of items for the care of appearance originating from the territory of Eurasia, made it possible to revise the point of view widespread in scientific circles on the functional purpose of the so-called bone "needle cases" from tubular bones of small animals and "needles / punctures" that were inside. In the course of the study, the items found in the cultural layer of the settlement sites of the Tobol-Irtysh region were classified as items for caring for the appearance - toothpicks placed in a case.

Protective Effect of Rho-Associated Protein Kinase (ROCK) Activated by Bone Marrow Mesenchymal Stem Cells on Bone Strength and Osteoblasts in Rats with Osteoporosis

This study assesses BMSCs’ effect on bone strength and osteoblast activity in rats with osteoporosis. 40 SD female rats were assigned into normal (A) group, model (B) group, puerarin (C) group, and BMSCs (D) group followed by analysis of bone strength by small animal bone strength analyzer and the expression of ROCK1 protein in tibia by immunohistochemistry. The bone strength of group B was lower than group A (P < 0.05), and higher in groups C and D than group B (P < 0.05) with further higher in group D than group C (P < 0.05). Group A showed obvious mesh cancellous bone trabecular bone and continuity, compact structure, and group B cells appeared loose and irregular distribution, parts cancellous bone trabeculae decrease and part of the fracture, and arranged in sparse irregular distribution. The decrease of trabecular bone in group C and D is not more obvious than group B and the morphology in group D got better improved than in group C. Group B showed lower proliferation of osteoblasts and ALP activity than group A (P < 0.05) and the proliferation and ALP activity in groups C and D was increased significantly compared with group B (P < 0.05). ROCK1 expression was significantly lower in group B than group A (P < 0.05) higher in groups C and D than group B (P < 0.05). Bone marrow mesenchymal stem cells (BMSCs) can effectively increase the bone strength and activity of osteoblasts in rats with osteoporosis, and promote the activation of ROCK signal.

Pulsed ultrasound for bone regeneration - outcomes and hurdles in the clinical application: a systematic review.

Impaired bone-fracture healing is associated with long-term musculoskeletal disability, pain and psychological distress. Low-intensity pulsed ultrasound (LIPUS) is a non-invasive and side-effect-free treatment option for fresh, delayed- and non-union bone fractures, which has been used in patients since the early 1990s. Several clinical studies, however, have questioned the usefulness of the LIPUS treatment for the regeneration of long bones, including those with a compromised healing. This systematic review addresses the hurdles that the clinical application of LIPUS encounters. Low patient compliance might disguise the effects of the LIPUS therapy, as observed in several studies. Furthermore, large discrepancies in results, showing profound LIPUS effects in regeneration of small-animal bones in comparison to the clinical studies, could be caused by the suboptimal parameters of the clinical set-up. This raises the question of whether the so-called "acoustic dose" requires a thorough characterisation to reveal the mechanisms of the therapy. The adequate definition of the acoustic dose is especially important in the elderly population and patients with underlying medical conditions, where distinct biological signatures lead to a delayed regeneration. Non-industry-funded, randomised, double-blind, placebo-controlled clinical trials of the LIPUS application alone and as an adjuvant treatment for bones with complicated healing, where consistent control of patient compliance is ensured, are required.

The antimicrobial activity and biocompatibility of a controlled gentamicin-releasing single-layer sol-gel coating on hydroxyapatite-coated titanium.

AimsThe aim of this study was to develop a single-layer hybrid organic-inorganic sol-gel coating that is capable of a controlled antibiotic release for cementless hydroxyapatite (HA)-coated titanium orthopaedic prostheses.MethodsCoatings containing gentamicin at a concentration of 1.25% weight/volume (wt/vol), similar to that found in commercially available antibiotic-loaded bone cement, were prepared and tested in the laboratory for: kinetics of antibiotic release; activity against planktonic and biofilm bacterial cultures; biocompatibility with cultured mammalian cells; and physical bonding to the material (n = 3 in all tests). The sol-gel coatings and controls were then tested in vivo in a small animal healing model (four materials tested; n = 6 per material), and applied to the surface of commercially pure HA-coated titanium rods.ResultsThe coating released gentamicin at > 10 × minimum inhibitory concentration (MIC) for sensitive staphylococcal strains within one hour thereby potentially giving effective prophylaxis for arthroplasty surgery, and showed > 99% elution of the antibiotic within the coating after 48 hours. There was total eradication of both planktonic bacteria and established bacterial biofilms of a panel of clinically relevant staphylococci. Mesenchymal stem cells adhered to the coated surfaces and differentiated towards osteoblasts, depositing calcium and expressing the bone marker protein, osteopontin. In the in vivo small animal bone healing model, the antibiotic sol-gel coated titanium (Ti)/HA rod led to osseointegration equivalent to that of the conventional HA-coated surface.ConclusionIn this study we report a new sol-gel technology that can release gentamicin from a bioceramic-coated cementless arthroplasty material. In vitro, local gentamicin levels are in excess of what can be achieved by antibiotic-loaded bone cement. In vivo, bone healing in an animal model is not impaired. This, thus, represents a biomaterial modification that may have the potential to protect at-risk patients from implant-related deep infection.Cite this article: Bone Joint J 2021;103-B(3):522–529.

The Role of Vitamin D in Small Animal Bone Metabolism.

Dogs and cats have differences in vitamin D metabolism compared to other mammalian species, as they are unable to perform vitamin D cutaneous synthesis through sun exposure. Therefore, they are dependent on the dietary intake of this nutrient. The classic functions of vitamin D are to stimulate intestinal calcium and phosphate absorption, renal calcium and phosphate reabsorption and regulate bone mineral metabolism. Thus, it is an important nutrient for calcium and phosphorus homeostasis. This review highlights the evidence of the direct and indirect actions of vitamin D on bone mineral metabolism, the consequences of nutritional imbalances of this nutrient in small animals, as well as differences in vitamin D metabolism between different size dogs.

Global and site-specific analysis of bone in a rat model of spinal cord injury-induced osteoporosis

Micro-Computed Tomography bone analysis is the gold standard method for assessing trabecular and cortical bone microarchitecture in small animal bones. This technique reports morphometric parameters as averages over selected volumes of interest (VOIs). This study proposes the introduction of an additional global 2D morphometric step into the analysis process, that provides a survey of the underlying morphometric variation present throughout both trabecular and cortical bone. The visualisation of these morphometric distributions provides a systematic approach to VOI selection that provides rationale and adds confidence to subsequent 3D morphometric analysis. To test the applicability and value of this methodological addition it was applied to the distal femur of a rat model of spinal cord injury (SCI)-induced osteoporosis. The 2D morphometric variation of both trabecular and cortical bone was quantified as a function of bone length. SCI-induced osteoporosis was localised in i) trabecular bone, where metaphyseal bone was more severely affected than epiphyseal bone, and there was a significant reduction in Distal Femoral Trabecular Extent, a new parameter defined here that quantifies how far trabecular bone penetrates in to the marrow cavity, ii) cortical bone, where diaphyseal bone underwent significant lowering of both cortical area and thickness, while distal-metaphyseal bone did not. Theses site-specific changes were validated, further elucidated and compared with follow-up conventional 3D analysis. The techniques applied here are equally applicable to other long bones (tibia, humerus, radius, ulna), other types of imaging modality and other types of experimental design including the effects of rehabilitation, aging, loading, gene knockout and pharmacological intervention.

Interpretation of the Wyoming Basin’s Holocene faunal record: deboning, transport, NISP/MNI estimates, and food yields

The paucity of artiodactyl bone at many Wyoming Basin archaeological sites is often explained by suggesting that bison and other large mammals were unavailable. Ignored is the ethnographic, historic, and modern record of deboning large animals at the kill site, and the transport of meat only to the residential camps. The resulting under-representation of artiodactyl bone skews the data base in favor of small animal bone, primarily from lagomorphs, which were likely brought into the residential camps whole. This problem is compounded when NISP data is used to infer subsistence, because it assigns each bone specimen, regardless of species, the same subsistence value. Consequently, available food yield based on average live weight is suggested to be a better estimate of the human diet, and consequent subsistence focus, than are interpretations based on the exclusive use of NISP/MNI.

A Morphometric and Cellular Analysis Method for the Murine Mandibular Condyle.

The temporomandibular joint (TMJ) has the capacity to adapt to external stimuli, and loading changes can affect the position of condyles, as well as the structural and cellular components of the mandibular condylar cartilage (MCC). This manuscript describes methods for analyzing these changes and a method for altering the loading of the TMJ in mice (i.e., compressive static TMJ loading). The structural evaluation illustrated here is a simple morphometric approach that uses the Digimizer software and is performed in radiographs of small bones. In addition, the analysis of cellular changes leading to alterations in collagen expression, bone remodeling, cell division, and proteoglycan distribution in the MCC is described. The quantification of these changes in histological sections - by counting the positive fluorescent pixels using image software and measuring the distance mapping and stained area with Digimizer - is also demonstrated. The methods shown here are not limited to the murine TMJ, but could be used on additional bones of small experimental animals and in other regions of endochondral ossification.

The fracture toughness of small animal cortical bone measured using arc-shaped tension specimens: Effects of bisphosphonate and deproteinization treatments

Small animal models, and especially transgenic models, have become widespread in the study of bone mechanobiology and metabolic bone disease, but test methods for measuring fracture toughness on multiple replicates or at multiple locations within a single small animal bone are lacking. Therefore, the objective of this study was to develop a method to measure cortical bone fracture toughness in multiple specimens and locations along the diaphysis of small animal bones. Arc-shaped tension specimens were prepared from the mid-diaphysis of rabbit ulnae and loaded to failure to measure the radial fracture toughness in multiple replicates per bone. The test specimen dimensions, crack length, and maximum load met requirements for measuring the plane strain fracture toughness. Experimental groups included a control group, bisphosphonate treatment group, and an ex vivo deproteinization treatment following bisphosphonate treatment (5 rabbits/group and 15 specimens/group). The fracture toughness of ulnar cortical bone from rabbits treated with zoledronic acid for six months exhibited no difference compared with the control group. Partially deproteinized specimens exhibited significantly lower fracture toughness compared with both the control and bisphosphonate treatment groups. The deproteinization treatment increased tissue mineral density (TMD) and resulted in a negative linear correlation between the measured fracture toughness and TMD. Fracture toughness measurements were repeatable with a coefficient of variation of 12–16% within experimental groups. Retrospective power analysis of the control and deproteinization treatment groups indicated a minimum detectable difference of 0.1MPa·m1/2. Therefore, the overall results of this study suggest that arc-shaped tension specimens offer an advantageous new method for measuring the fracture toughness in small animal bones.

Accuracy and repeatability of long-bone replicas of small animals fabricated by use of low-end and high-end commercial three-dimensional printers.

OBJECTIVE To assess the repeatability and accuracy of polymer replicas of small, medium, and large long bones of small animals fabricated by use of 2 low-end and 2 high-end 3-D printers. SAMPLE Polymer replicas of a cat femur, dog radius, and dog tibia were fabricated in triplicate by use of each of four 3-D printing methods. PROCEDURES 3-D renderings of the 3 bones reconstructed from CT images were prepared, and length, width of the proximal aspect, and width of the distal aspect of each CT image were measured in triplicate. Polymer replicas were fabricated by use of a high-end system that relied on jetting of curable liquid photopolymer, a high-end system that relied on polymer extrusion, a triple-nozzle polymer extrusion low-end system, and a dual-nozzle polymer extrusion low-end system. Polymer replicas were scanned by use of a laser-based coordinate measurement machine. Length, width of the proximal aspect, and width of the distal aspect of the scans of replicas were measured and compared with measurements for the 3-D renderings. RESULTS 129 measurements were collected for 34 replicas (fabrication of 1 large long-bone replica was unsuccessful on each of the 2 low-end printers). Replicas were highly repeatable for all 3-D printers. The 3-D printers overestimated dimensions of large replicas by approximately 1%. CONCLUSIONS AND CLINICAL RELEVANCE Low-end and high-end 3-D printers fabricated CT-derived replicas of bones of small animals with high repeatability. Replicas were slightly larger than the original bones.

Discrete tomography in an in vivo small animal bone study.

This study aimed at assessing the feasibility of a discrete algebraic reconstruction technique (DART) to be used in in vivo small animal bone studies. The advantage of discrete tomography is the possibility to reduce the amount of X-ray projection images, which makes scans faster and implies also a significant reduction of radiation dose, without compromising the reconstruction results. Bone studies are ideal for being performed with discrete tomography, due to the relatively small number of attenuation coefficients contained in the image [namely three: background (air), soft tissue and bone]. In this paper, a validation is made by comparing trabecular bone morphometric parameters calculated from images obtained by using DART and the commonly used standard filtered back-projection (FBP). Female rats were divided into an ovariectomized (OVX) and a sham-operated group. In vivo micro-CT scanning of the tibia was done at baseline and at 2, 4, 8 and 12weeks after surgery. The cross-section images were reconstructed using first the full set of projection images and afterwards reducing them in number to a quarter and one-sixth (248, 62, 42 projection images, respectively). For both reconstruction methods, similar changes in morphometric parameters were observed over time: bone loss for OVX and bone growth for sham-operated rats, although for DART the actual values were systematically higher (bone volume fraction) or lower (structure model index) compared to FBP, depending on the morphometric parameter. The DART algorithm was, however, more robust when using fewer projection images, where the standard FBP reconstruction was more prone to noise, showing a significantly bigger deviation from the morphometric parameters obtained using all projection images. This study supports the use of DART as a potential alternative method to FBP in X-ray micro-CT animal studies, in particular, when the number of projections has to be drastically minimized, which directly reduces scanning time and dose.

Microtomography with sandwich detectors for small-animal bone imaging

An x-ray radiographic system consisting of two detectors in tandem, or a sandwich detector, can produce dual-energy image from a single-shot exposure. Subtraction of two images obtained from the two detectors can produce a sharper image through an unsharp masking effect if the two images are formed at different spatial resolutions. This is indeed possible by incorporating different thicknesses of x-ray conversion layers in the detectors. In this study, we have developed a microtomography system with a sandwich detector in pursuit of high-resolution bone-enhanced small-animal imaging. The results show that the bone-enhanced images reconstructed from the dual-energy projection data provide higher visibility of bone details than the conventionally reconstructed images. The microtomography with the single-shot dual-energy sandwich detector will be useful for the high-resolution bone-enhanced small-animal imaging.

The scene of a spectacular feast (part II): Animal remains from Dolní Věstonice II, the Czech Republic

Studies of the archaeological and paleontological materials from Dolní Věstonice II provide insight into the lives of humans nearly 30,000 years ago. Bones of small animals (birds, hares, foxes) and medium animals (wolf, reindeer, wolverine) dominate at the site, but there are also bones of large mammals (bears, cave lion, horse and mammoth), showing that the prey spectrum of the hunters was wide. The large total number of animal remains supports the suggestion that these were accumulated during a relatively long human occupation of the site, perhaps lasting months or even years.In Moravia early Gravettian hunting strategy was less specialized than in later periods (e.g., at Milovice I). Large (mammoth, horse), medium (reindeer), and small animals (birds and hares) were important components of the diet and also used as raw materials. The carnivores – wolf, wolverine, foxes – were certainly important prey, not only for their hides but also for use in tool production and the creation of ornaments. Pavlovians also hunted even the larger carnivores (bears and lion).

Who eats whom? Taphonomic analysis of the avian record from the Middle Paleolithic site of Teixoneres Cave (Moià, Barcelona, Spain)

Small animal bones, such as those of birds, are commonly found at many archaeological sites framed in the early Late Pleistocene. Teixoneres Cave, on the Iberian Peninsula, is one of these, and includes evidence of Neanderthal activities involving large game and, sporadically, smaller prey such as rabbits. Here, we present data from the avian assemblage recovered from this site, which is mainly comprised of specimens from the Corvidae and Phasianidae families. In order to determine which predators (hominins, mammalian carnivores and/or raptors) contributed to this avian accumulation, the general occupational dynamics within the site must first be understood. To this end, the bird remains obtained from the four main subunits excavated to date (IIa, IIb, IIIa, IIIb) have been analyzed from a taphonomic perspective. Our results show that the birds at the site mainly originated from non-hominin input episodes. While the activity of nocturnal raptors was found throughout the sequence, the activity of mammalian carnivores seems to be more intense in specific archaeological units. We compared the data yielded by our study with other data from the site, reinforcing the general position that hominins made use of the cave during short-term occupations, which alternated with predator use.

Synchrotron radiation X-ray microtomography and histomorphometry for evaluation of chemotherapy effects in trabecular bone structure

Three-dimensional microtomography has the potential to examine complete bones of small laboratory animals with very high resolution in a non-invasive way. One of the side effects caused by some chemotherapy drugs is the induction of amenorrhea, temporary or not, in premenopausal women, with a consequent decrease in estrogen production, which can lead to bone changes. In the present work, the femur heads of rats treated with chemotherapy drugs were evaluated by 3D histomorphometry using synchrotron radiation microcomputed tomography. Control animals were also evaluated for comparison. The 3D tomographic images were obtained at the SYRMEP (SYnchrotron Radiation for MEdical Physics) beamline at the Elettra Synchrotron Laboratory in Trieste, Italy. Results showed significant differences in morphometric parameters measured from the 3D images of femur heads of rats in both analyzed groups.

제주도 종달리 1819번지 패총에서 출토된 뼈 유물의 분류

Animal bones excavated with earthenware from the shell mound at the Jeju Jongdali 1819 archeological site, where three consecutive chronological layers covering the Neolithic (B.C. 15C-B.C. 10C), early Tamra, and late Tamra periods have been identified, were morphologically classified. The majority of the bones from all three periods were broken or split. The major fauna of the mammalian bones in all periods were Cervus spp., Sus scrofa, and Bos taurus. In the early and late Tamra periods, bones of small animals including Mustela sibirica coreana, Meles meles, Rodentia, and Aves were also found in small number. The excavated bones were from all parts of the animal bodies, including head, trunk, forelimb, and hindlimb. Collectively, these findings suggest that the major fauna from the Neolithic to late Tamra periods consisted of Cervus spp., Sus scrofa, and Bos taurus and that the fauna was dissected and carried to the shell mound site after hunting. Information from the bone remains in the shell mound are useful data for study of the wildlife and domestic animals living during the prehistoric period of Jeju Island.

A Novel Videography Method for Generating Crack-Extension Resistance Curves in Small Bone Samples

Assessment of bone quality is an emerging solution for quantifying the effects of bone pathology or treatment. Perhaps one of the most important parameters characterising bone quality is the toughness behaviour of bone. Particularly, fracture toughness, is becoming a popular means for evaluating bone quality. The method is moving from a single value approach that models bone as a linear-elastic material (using the stress intensity factor, K) towards full crack extension resistance curves (R-curves) using a non-linear model (the strain energy release rate in J-R curves). However, for explanted human bone or small animal bones, there are difficulties in measuring crack-extension resistance curves due to size constraints at the millimetre and sub-millimetre scale. This research proposes a novel “whitening front tracking” method that uses videography to generate full fracture resistance curves in small bone samples where crack propagation cannot typically be observed. Here we present this method on sharp edge notched samples (<1 mm×1 mm×Length) prepared from four human femora tested in three-point bending. Each sample was loaded in a mechanical tester with the crack propagation recorded using videography and analysed using an algorithm to track the whitening (damage) zone. Using the “whitening front tracking” method, full R-curves and J-R curves could be generated for these samples. The curves for this antiplane longitudinal orientation were similar to those found in the literature, being between the published longitudinal and transverse orientations. The proposed technique shows the ability to generate full “crack” extension resistance curves by tracking the whitening front propagation to overcome the small size limitations and the single value approach.

Worked Human Femur from Gohar Tepe, Iran

A simple tool made from a gracile human femoral shaft was retrieved from a small animal bone assemblage found in a Late Bronze Age stratum at Gohar Tepe, Iran. The specimen has been identified as a chisel or gouge for which no analogous examples are known in the Near East. Studies examining similar tools from other regions suggest that such a tool may have been used for wood processing or pottery smoothing. Copyright © 2012 John Wiley & Sons, Ltd.

Mechanistic modeling of a nanoscratch test for determination of in situ toughness of bone

The objective of this study was to develop a nanoscratch technique that can be used to measure the in situ toughness of bone at micro/nanostructural levels. Among the currently possible techniques, the surface scratch test may be conducted on very small regions, thus exhibiting a potential in determining the in situ failure behavior of materials. To adapt such a technique for assessing bone toughness at the micro/nanostructural levels and for limited stocks in small animal bone models (e.g. zebra finish and mice), a simple but reasonably accurate mechanistic model for the nanoscratch test was developed in this study. This model was based on the assumption that the removal energy of the tissue required during the nanoscratch test is the manifestation of the in situ toughness and the shear flow stress during the removal process is a measure of the in situ strength of bone. In addition, the experimental methodologies were developed to determine the elastic recovery force and frictional coefficients between the scratch tip and bone specimens that are required by the model. Finally, the efficacy of the nanoscratch technique was verified by testing bone samples from control (wild type), mild, and severe osteogenesis imperfecta (OI) mice, which have a distinct degree of brittleness. The experimental results indicated that the nanoscratch test could sensitively detect the in situ brittleness and strength of bone from the animal models.

Small animal bone healing models: Standards, tips, and pitfalls results of a consensus meeting

Small animal fracture models have gained increasing interest in fracture healing studies. To achieve standardized and defined study conditions, various variables must be carefully controlled when designing fracture healing experiments in mice or rats. The strain, age and sex of the animals may influence the process of fracture healing. Furthermore, the choice of the fracture fixation technique depends on the questions addressed, whereby intra- and extramedullary implants as well as open and closed surgical approaches may be considered. During the last few years, a variety of different, highly sophisticated implants for fracture fixation in small animals have been developed. Rigid fixation with locking plates or external fixators results in predominantly intramembranous healing in both mice and rats. Locking plates, external fixators, intramedullary screws, the locking nail and the pin-clip device allow different degrees of stability resulting in various amounts of endochondral and intramembranous healing. The use of common pins that do not provide rotational and axial stability during fracture stabilization should be discouraged in the future. Analyses should include at least biomechanical and histological evaluations, even if the focus of the study is directed towards the elucidation of molecular mechanisms of fracture healing using the largely available spectrum of antibodies and gene-targeted animals to study molecular mechanisms of fracture healing. This review discusses distinct requirements for the experimental setups as well as the advantages and pitfalls of the different fixation techniques in rats and mice.