Patient-specific bone modelling and remodelling simulation of hypoparathyroidism based on human iliac crest biopsies

We previously developed a load-adaptive bone modelling and remodelling simulation model that can predict changes in the bone micro-architecture as a result of changes in mechanical loading or cell activity. In combination with a novel algorithm to estimate loading conditions, this offers the possibility for patient-specific predictions of bone modelling and remodelling. Based on such models, the underlying mechanisms of bone diseases and/or the effects of certain drugs and their influence on the bone micro-architecture can be investigated. In the present study we test the ability of this approach to predict changes in bone micro-architecture during hypoparathyroidism (HypoPT), as an illustrative example. We hypothesize that, apart from reducing bone turnover, HypoPT must also lead to increased osteocyte mechanosensitivity in order to explain the changes in bone mass seen in patients. Healthy human iliac crest biopsies were used as the starting point for the simulations that mimic HypoPT conditions and the resultant micro-architectures were compared to age-matched clinical HypoPT biopsies. Simulation results were in good agreement with the clinical data when osteocyte mechanosensitivity was increased by 40%. In conclusion, the results confirm our hypothesis, and also demonstrate that patient-specific bone modelling and remodelling simulations are feasible.     

Effectiveness of fresh frozen and cryopreserved homologue iliac crest grafts used in sinus lifting: a comparative study

In the last decade, several investigators have reported that autologous and homologous fresh frozen bones (FFB) are effective materials to restore alveolar ridges previous to insert dental implants. Recently we have used cryopreserved homologue grafts (CFFB). Here we reported a retrospective comparative study between implants inserted in FFB and CFFB evaluate their clinical outcome. Patients were treated with a split mouth scheme for bone grafting with FFB and CFFB and spiral family implants (SPI) were inserted in the same surgical time. Several variables (patient, grafts, anatomic site, implant, prosthetic restoration) were investigated. Implant’ failure and peri-implant bone resorption were considered as predictor of clinical outcome. A total of 84 SFIs were inserted in 12 patients. Implants were inserted to replace 8 incisors, 4 cuspids, 31 premolars and 41 molars. The mean follow-up was 14 months. Three out of 84 implants was lost (i.e. survival rate SVR = 96.4%) and no differences were detected among the studied variables. Similar result was obtained by analyzing the crestal bone resorption around implant’ neck (i.e. success rate). FFB and CFFB have high and comparable survival and success rate. Implants inserted with one step surgical procedure in native (i.e. not grafted) bone, FFB and CFFB have similar clinical outcome.     

Small free vascularized iliac crest bone grafts in reconstruction of the scaphoid bone: a retrospective study in 60 cases.

Carpal instability may result in progressive degenerative arthritis of the wrist. The surgical goal of the reconstruction of scaphoid nonunion is to achieve bone union and to restore the scaphoid. Many procedures are described to treat scaphoid nonunion for different indications. This retrospective study reports on the anatomical fundamentals, the operative procedure, and the results of 60 patients (21 with recalcitrant scaphoid nonunion that lasted longer than 4 years, 26 with an avascular pole fragment, and 13 with scaphoid nonunion after previous surgery) who were treated by a small free vascularized iliac crest bone graft. All 60 patients have routinely been followed up clinically and with magnetic resonance imaging. Union was achieved in 91.7 percent by improvement of stability and the compromised vascularity of the scaphoid. The bone flap loss rate and persisting nonunion was 8.3 percent, leading to progressive arthritis and carpal collapse. Complaints concerning discomforts caused by the scar were heard from 40.1 percent of the patients, and 31.7 percent complained of discomforts caused by the bony deformity. Bone deformations on the donor site were detected radiologically in 63.3 percent of the patients. In 31.7 percent, an impairment of the lateral femoral cutaneous nerve was noted. Reconstruction of the scaphoid by means of implantation of a vascularized iliac bone graft proved efficient to treat avascular recalcitrant scaphoid nonunion and pseudarthrosis with avascular proximal pole fragments.     

Revascularization of cranial versus iliac crest bone grafts in the rat

Full-thickness cranial (membranous) and split-thickness iliac crest (endochondral) onlay bone grafts were placed subperiosteally without fixation onto the snout (membranous) and tibia (endochondral) in 30 rats. The animals had been divided into three equal groups in which the bone grafts had been demineralized, autoclaved, or used fresh. Recipient sites were harvested at 7 and 14 days at the snout and 14 days at the tibia, and revascularization was studied utilizing silicone rubber injection and a gridcounting technique. Endochondral grafts were found to have quantitatively greater revascularization than membranous grafts in all three groups at both sites (p less than 0.005). There was generally no statistically significant difference in revascularization between fresh and demineralized grafts, but vessel ingrowth was significantly decreased in autoclaved implants as compared with fresh grafts. Differences in graft architecture are theorized to account for the disparity in revascularization in endochondral and membranous grafts. Angiogenic and chemotactic factors are thought to play a role in the similarities and differences in revascularization among fresh, demineralized, and autoclaved bone.     

Comparison of phenotypic markers and neural differentiation potential of human bone marrow stromal cells from the cranial bone and iliac crest

Cellular therapies represent a new frontier in the treatment of neurological diseases. Accumulating evidence from preclinical studies of animal models suggests that mesenchymal stromal cells (MSCs), also known as mesenchymal stem cells, are an effective therapy for neurological diseases. In this study, we established human MSC lines from both cranial bone marrow (cBMMSCs) and iliac crest bone marrow (iBMMSCs) from the same donors and found that cBMMSCs show higher expression of neural crest-associated genes than iBMMSCs. Moreover, as observed in both mRNA and protein assays, neurogenic-induced cells from cBMMSCs expressed significantly higher levels of neural markers, such as NESTIN, SLUG, SOX9, and TWIST, than those from iBMMSCs. Thus, cBMMSCs showed a greater tendency than iBMMSCs to differentiate into neuron-like cells.     

An experimental and theoretical approach of elasticity and viscoelasticity of compact and spongy bone with periodic homogenization

An experimental study on samples from a fresh human femur highlighted the viscoelastic behaviour of bone. The memory aspect of the behaviour is weak compared to the elastic aspect. A theoretical and numerical approach based on the finite element method illustrates the existing bond between the structure and the micro behaviour on the one hand, and the macro behaviour on the other. The orthotropic elastic behaviour observed by various authors is shown numerically. The memory functions of compact and spongy bone are given. The numerical results illustrate the behaviour observed in the experiments.     

Sinusoids of the bone marrow stroma in the human iliac bone

Results of histological and ultramicroscopic investigations of sinusoid vessels are presented. Interrupted structure of the basal membrane and continuous endothelial lining are revealed. Endotheliocytes of two types are described. At places of blood cells migration functional activity of endothelium is increased and the basal membrane is absent. A suggestion is made that the endothelial cells directly influence hemopoiesis.     

Endosteal cells of the bone marrow stroma in human iliac bone

The data on histological and electron microscopical investigation of the endosteal cells of the human iliac bone are presented. Three types of stromal elements in the endosteum, differing in their ultrastructural organization have been revealed. In the endosteal areas young hemopoietic cells are present, they are closely connected with the stroma. A suggestion is made on an important role of the endosteum in processes of proliferation and differentiation of hemopoietic predecessors.     

Blood supply of the compact and spongy myocardium of fish, amphibia and reptiles]

Coronal arteries were injected with lead carbonate suspension and with Indian ink and cleared preparations 150--300 mkm thick were made in 195 hearts of fish, amphibians and reptiles and studied roentgenographically. It was stated that in Chondrichthyes (shark, skate) and in Chondrostei (beluga, stellate sturgeon, sturgeon), as well as in alligator both compact and spongy myocardium of the cardiac ventricle possess blood vessels. In teleostei, amphibians and reptiles (except alligator) spongy myocardium is avascular and receives its nutrition from the ventricle. In view of the data on the presence of blood vessels in the spongy myocardium in some vertebrates, it is impossible to accept the theory suggested by Grant and Regnier according to which vessels in the heart walls appear only in connection with compactization of the myocardium. Vascularization of the spongy myocardium is closely connected with oxygen saturation of the blood flowing through the heart. When this saturation is not satisfactory, the spongy myocardium has blood vessels. In alligator, vascularization of the spongy myocardium is connected with the fact that the heart has four chambers and there are arterial and venous blood streams.     

Targeted regeneration of bone in the osteoporotic human femur

We have recently developed image processing techniques for measuring the cortical thicknesses of skeletal structures in vivo, with resolution surpassing that of the underlying computed tomography system. The resulting thickness maps can be analysed across cohorts by statistical parametric mapping. Applying these methods to the proximal femurs of osteoporotic women, we discover targeted and apparently synergistic effects of pharmaceutical osteoporosis therapy and habitual mechanical load in enhancing bone thickness.     

Adipocytes and reticular cells of the bone marrow stroma in the human iliac bone

The results of the histological and electron microscopic investigation of adipose and reticular cells and their interconnections with blood cells are presented in the material of trephine biopsies of the iliac bone. A possibility for development of adipocytes from the adventitial reticular cells is demonstrated. Close contacts are revealed between pre-adipocytes and young hemopoietic cells. Two types of the reticular cells are characterized, they differ in their position, structural organization and interconnection with the young hemopoietic elements. The peculiarities revealed in the morphofunctional state of the microenvironmental structures demonstrate functional variegation of the stromal elements, and also attest an essential importance of intercellular contacts of the hemopoietic predecessors and the stromal cells in maintaining the hemopoietic function of the bone marrow.     

Age-related and gender-related differences between human vertebral and iliac crest bone--a histomorphometric study on the population of the Mediterranean Coast of Croatia

In this study, osseous tissue was examined in normal adult population that has inhabited areas by the Croatian Adriatic Sea. The most of such studies have shown that women are prone to lose bone connectedness, while men are predisposed to be a stronger constitution and they start with greater bone mass, though. Bone samples from two different anatomic sites were analyzed. The crista iliaca and the lumbar vertebra represent functionally different organs too. We wanted to consider weather the same age- and gender-related changes affect these two organs due to normal aging. Static histomorphometry was used to quantify involution changes in the trabecular bone. Results showed that involution process more severely affects women than men. Age-related structural changes were more prominent in lumbar vertebra than in iliac crest bone. Severe structural changes in lumbar vertebra could subsequently lead to a dysfunctional and deformed vertebral column. Therefore, iliac crest bone biopsies could hardly explain involution process that affects lumbar spine.     

Density of fresh and embalmed human compact and cancellous bone

This investigation was undertaken to determine the densities of fresh human compact and cancellous bone, with its marrow constituents intact, and compact and cancellous bone taken from embalmed cadavers. The density of 107 bone specimens was determined on the basis of the weight and volume of each specimen. Mean density results were: (1) fresh compact bone, 1.85; (2) fresh cancellous bone, 1.08; (3) embalmed compact bone, 1.85; and (4) embalmed cancellous bone, 1.09. Embalming did not appear to have altered the density of either compact or cancellous bone. It was observed also that the densities of several bone specimens taken from the same area of one bone varied greatly.     

Human mesenchymal progenitor cells derived from alveolar bone and human bone marrow stromal cells: a comparative study

The aim of the present study was to evaluate the potential of intraoral harvested alveolar bone as an alternative source of multipotent mesenchymal stromal cells for future applications in oral and maxillofacial tissue engineering. Explant cultures were established from 20 alveolar bone samples harvested from the oblique line immediately before wisdom tooth removal. Morphology and proliferation characteristics of the in vitro expanded cells, referred to as human alveolar bone-derived cells (hABDCs), were studied using phase-contrast microscopy. Immunocytochemical analysis of their surface marker expression was conducted using monoclonal antibodies defining mesenchymal stromal cells. To evaluate their multilineage differentiation potential, hABDCs were induced to differentiate along the osteogenic, adipogenic, and chondrogenic lineage and compared to bone marrow mesenchymal stromal cells (hBMSCs) on mRNA and protein levels applying RT-PCR and cytochemical staining methods. hABDCs showed typical morphological characteristics comparable to those of hBMSCs such as being mononuclear, fibroblast-like, spindle-shaped, and plastic adherent. Immunophenotypically, cells were positive for CD105, CD90, and CD73 while negative for CD45, CD34, CD14, CD79α, and HLA-DR surface molecules, indicating an antigen expression pattern considered typical for multipotent mesenchymal stromal cells. As evidenced by RT-PCR and cytochemistry, hABDCs showed multilineage differentiation and similar chondrogenic and osteogenic differentiation potentials when compared to hBMSCs. Our findings demonstrate that human alveolar bone contains mesenchymal progenitor cells that can be isolated and expanded in vitro and are capable of trilineage differentiation, providing a reservoir of multipotent mesenchymal cells from an easily accessible tissue source.     

Degradation of biomolecules: a comparative study of diagenesis of DNA and proteins in human bone tissue

Diagenesis of macromolecules is a not yet fully understood process that can be important for anthropological and forensic research. Trying to elucidate the diagenesis of DNA and proteins we investigated the process of fragmentation of DNA and razemisation of aspartic acid in human bone material. We created an in vitro-model of accelerated aging by incubating bone samples in hot water. A comparison of diagenesis of molecules in those artificially aged samples with altogether 30 historical bones from different regions and of different ages was carried out. The in vitro-model showed the expected positive correlation between the increase of razemisation of aspartic acid and DNA fragmentation, while there was a much lesser correlation when investigating historical bones. The in vitro-model showed the expected correlation between the increase of razemisation of aspartic acid and DNA fragmentation and to a much lesser extent in historical bones. This study shows that diagenesis is probably influenced by additional forces affecting different macromolecules in different ways.     

Biochemical, histochemical and immunohistochemical study of glycosaminoglycans in human meningiomas

The localization and quantitation of glycosaminoglycans classes (GAGs) were studied in human meningiomas. Meningiomas presented high amounts of these compounds and electrophoretic separation revealed that they were 90% sulphated. The Alcian method and a polyclonal antiserum against chondroitin sulphate were used to localize the different GAGs in tissue sections. Quantitative and qualitative differences and different tissue distributions of GAGs were observed among transitional, syncytial and fibroblastic meningiomas. Syncytial meningiomas presented the lowest amount of GAGs and the immuno- and histochemical studies showed that they were located only in vessels and connectival trabeculae. Transitional meningiomas contained the highest concentration of GAGs; the percentage of the different GAG classes was similar to that observed in the syncytial oncotype indicating a quantitative but not qualitative difference between the two oncotypes. The high amount of GAGs in transitional meningiomas was attribute to the whorls, the structures stained by the histochemical and immunohistochemical techniques. The tumoral parenchyma of these two oncotypes was negative. On the contrary, fibroblastic meningiomas showed a fine meshwork among tumoral cells containing chondroitin sulphate and heparan sulphate. Biochemical data were consistent with the histochemical and immunohistochemical findings revealing a high percentage of chondroitin sulphate and heparan sulphate in fibroblastic meningiomas. This study suggests that the three meningioma types have different abilities to produce extracellular matrix components.     

Mathematical modelling of trabecular bone structure: the evaluation of analytical and quantified surface to volume relationships in the femoral head and iliac crest

The three-dimensional architecture of trabecular bone has structural trends related to physical function as described by Wolff's law. Mathematical modelling provides a means of analysing these structures through the use of simplified representations. A single measure of mineralized bone volume per unit volume of structure (Vv) and the surface area of mineralized bone per unit volume of structure (Sv) does not identify a particular architecture in any detail; the way in which Sv changes in relation to Vv does provide this information as the structure remodels. A series of structures using the elements of plates and rods were created. The rates of change of Sv with respect to Vv for trabecular structures give insight into differences in such models. Structures in the femoral head and iliac crest were analysed by power curve regression. In the principal compressive region, just above the medial cortex, advanced osteoarthritis was associated with a preferential loss of rods from the normal trabecular structure, resulting in a more plate-like architecture. The iliac crest remodelling that takes place in the osteoporotic appears to be the result of a generalised bone loss with some of the thinner elements of the structure being removed completely, resulting in an increase in unit cell dimension. The consequence of changing unit cell size has a major impact on surface availability for osteoblastic and osteoclastic activity. The simple plate model as a basis for the stereological analysis of trabecular structures is therefore limited because of the mixed plate and rod nature of trabecular architecture.     

Surface anatomy of branchial food traps of tadpoles: A comparative study

Branchial food traps are regions of specialized secretory tissue in the tadpole pharynx, where suspended food particles are trapped in mucus. Light and scanning electron microscopy were used to study branchial food traps from larvae of ten anuran families (36 species). Most anuran larvae from “advanced” (suborder Neobatrachia) families (e.g., Hylidae, Ranidae, Bufonidae) have distinct secretory pits at the posterior margins of the branchial food traps and secretory ridges elsewhere on these surfaces. The apices of columnar PAS-positive, secretory cells are exposed on the floors of the secretory pits or in rows at the tops of the secretory ridges (secretory zone). Tadpoles from most “archaic” (suborder Archaeobatrachia) families (Ascaphidae, Discoglossidae and Pelobatidae) either lack secretory pits, or have them poorly defined. They also lack secretory ridges but have columnar, mucus-secreting cells whose apices are exposed in a seemingly random fashion in the branchial food traps. Rhinophrynus (Archaeobatrachia: Rhinophrynidae) has secretory ridges, but the apices of secretory cells are not arranged in rows at the tops of the ridges; instead they erupt singly or in small clusters on the epithelial surface, in a pattern similar to that in Ascaphus, the discoglossids and the pelobatids. It is proposed that the generalized condition for the branchial food trap mucosa is one where the apices of secretory cells are exposed haphazardly on a flat epithelium and the derived condition is one where the surface is organized into ridges. The morphology of the branchial food traps in Rhinophrynus suggests that, phylogenetically, ridges preceded the coalescing of secretory cell apices into distinct rows. Pipidae and Microhylidae have unique patterns in the gross and microanatomy of their branchial food traps specific to their families. Branchial food trap morphology relates to diets of tadpoles as well as to taxonomy. Obligate macrophagous (e.g., carnivorous) tadpoles, irrespective of family, tend to have reduced branchial food traps, regularly lack secretory ridges and, in extreme cases, lack columnar mucus-secreting cells. Obligate microphagous forms (midwater suspension feeding of Xenopus, microhylids and Agalychnis), have straight parallel secretory ridges with narrow secretory zones and shallow troughs between the ridges. Secretory ridges may help to form mucus strands in which food particles are trapped, but they are not essential for planktonic entrapment. The hydrodynamic implications of the various topographic patterns remain unclear.