Newly Formed Bone Induced by Recombinant Human Bone Morphogenetic Protein-2: A Histological Observation.

PURPOSE: To observe, histologically, bone induced by recombinant human bone morphogenetic protein-2 (rhBMP-2) in onlay grafted and sinus lifted alveolaris. MATERIAL AND METHODS: Eighteen patients were treated with rhBMP-2 at concentration 1.5 mg/mL with an absorbable collagen sponge (ACS). The treated bone was harvested with small trephine bur at 5 or 7 months after surgery for the micro Computer Scanning (CT) and light microscopic observation. RESULTS: Micro CT showed clearly 3-dimensional trabecular bone structure. New bone formation and bone marrow structure were observed in the observed area. Osteoblastic cells existed along the new bone, and osteopontin was localized in the bone matrix weakly. In the connective tissue around the new bone, many CD34-positive blood vessel cells were present. Some tartrate-resistant acid phosphatase (TRAP)-positive osteoclastic cells were observed around bone at this stage. CONCLUSION: The application of rhBMP-2 with ACS induced a new bone accompanied by blood vessels in atrophied alveolaris. This suggests that rhBMP-2 is capable of osteoinductivity in human jaw.

[Histomorphometric evaluation of ridge preservation after molar tooth extraction].

OBJECTIVE: To evaluate bone formation in human extraction sockets with absorbed surrounding walls augmented with Bio-Oss and Bio-Gide after a 6-month healing period by histologic and histomorphometric analyses. METHODS: Six fresh molar tooth extraction sockets in 6 patients who required periodontally compromised moral tooth extraction were included in this study. The six fresh extraction sockets were grafted with Bio-Oss particle covered with Bio-Gide. The 2.8 mm×6.0 mm cylindric bone specimens were taken from the graft sites with aid of stent 6 months after the surgery. Histologic and histomorphometric analyses were performed. RESULTS: The histological results showed Bio-Oss particles were easily distinguished from the newly formed bone, small amounts of new bone were formed among the Bio-Oss particles, large amounts of connective tissue were found. Intimate contact between the newly formed bone and the small part of Bio-Oss particles was present. All the biopsy cylinders measurement demonstrated a high inter-individual variability in the percentage of the bone, connective tissues and Bio-Oss particles. The new bone occupied 11.54% (0-28.40%) of the total area; the connective tissues were 53.42% (34.08%-74.59%) and the Bio-Oss particles were 35.04% (13.92%-50.87%). The percentage of the particles, which were in contact with bone tissues, amounted to 20.13% (0-48.50%). CONCLUSION: Sites grafted with Bio-Oss particles covered with Bio-Gide were comprised of connective tissues and small amounts of newly formed bone surrounding the graft particles.

A Single Chance to Contact Multiple Targets: Distinct Osteocyte Morphotypes Shed Light on the Cellular Mechanism Ensuring the Robust Formation of Osteocytic Networks.

The formation of the complex osteocytic network relies on the emission of long cellular processes involved in communication, mechanical strain sensing, and bone turnover control. Newly deposited osteocytic processes rapidly become trapped within the calcifying matrix, and, therefore, they must adopt their definitive conformation and contact their targets in a single morphogenetic event. However, the cellular mechanisms ensuring the robustness of this unique mode of morphogenesis remain unknown. To address this issue, we examined the developing calvaria of the amphibian Xenopus tropicalis by confocal, two-photon, and super-resolution imaging, and described flattened osteocytes lying within a woven bone structured in lamellae of randomly oriented collagen fibers. While most cells emit peripheral and perpendicular processes, we report two osteocytes morphotypes, located at different depth within the bone matrix and exhibiting distinct number and orientation of perpendicular cell processes. We show that this pattern is conserved with the chick Gallus gallus and suggest that the cellular microenvironment, and more particularly cell-cell contact, plays a fundamental role in the induction and stabilization of osteocytic processes. We propose that this intrinsic property might have been evolutionarily selected for its ability to robustly generate self-organizing osteocytic networks harbored by the wide variety of bone shapes and architectures found in extant and extinct vertebrates.

Specialized connective tissue: bone, the structural framework of the upper extremity.

Bone is a connective tissue containing cells, fibers, and ground substance. There are many functions in the body in which the bone participates, such as storing minerals, providing internal support, protecting vital organs, enabling movement, and providing attachment sites for muscles and tendons. Bone is unique because its collagen framework absorbs energy, whereas the mineral encased within the matrix allows bone to resist deformation. This article provides an overview of the structure and function of bone tissue from a macroscopic to microscopic level and discusses the physiological processes contributing to upper extremity bone health. It concludes by discussing common conditions influencing upper extremity bone health.

The structure of cortical bone as revealed by the application of methods for the calcified matrix study.

Calcination and decalcification are basic procedures useful to a morphological approach of a biological, composite material like cortical bone. The study was carried out on a whole human femur conserved in liquid (from an educational collection). Cortical fracturing and SEM observation of vascular canals surface collagen texture was used to study bone deproteination at scalar temperatures (400-1,200°C) and acid bone decalcification at crescent time intervals. Heating burned and vaporized the organic matrix with shrinkage of the bone specimens as documented by the weight loss and transverse surface morphometry. SEM showed a pattern of aligned spherulites at 400°C which maintained the collagen fibrils layout (like a mineral cast), followed by a spherulites fusion progression with the temperature increments. At 1200°C a crystalline-like structure of tightly-packed trapezohendron units. XRD analysis supported the SEM morphology displaying the complete Debey rings of hydroxyapatite and spotted Debey rings of withlockite. Surface Ca and P elution was documented after 12 hr of exposition to the acid solution by dissolution of spherulites and the whole canal surface decalcified in depth after 15 days by SEM-EDAX analysis. The periodic pattern of collagen fibrils was still evident up to 15 days of decalcification together with fine granular deposits of a not-collagenic proteic material, while after 30 days no period was observed in the decalcified fibrils. Collagen mineral cast at 400°C calcination. Complete crystalline transformation at 1200°C. Up to 15 days of decalcification fibrils period maintained.

Assessment of infrazygomatic bone depth for mini-screw insertion.

OBJECTIVE: To investigate the bone depth at the infrazygomatic crest with regard to orthodontic mini-screw insertion. MATERIAL AND METHODS: Twenty-nine adult human dry skulls were imaged using CBCT technology, slice data were generated and multiple measurements were undertaken at three sites associated with the infrazygomatic crest and five different measurement levels. The data were analyzed using intraclass correlation and repeated measures ANOVA. RESULTS: The greatest bone depth was available at, on average, 11.48+/-1.92 mm apical from the cemento-enamel junction of the maxillary first molar and decreased rapidly further apically. Maximum bone depth (7.05+/-3.7 mm) was present at the lowest measurement level. However, here, insufficient clearance to the molar roots was present. Both the measurement site and the level at which the measurements were conducted had a significant impact on bone depth. CONCLUSIONS: When inserting orthodontic mini-screws (6 mm or longer) into the infrazygomatic crest while staying clear of the molar roots perforation of the maxillary sinus or the nasal cavity can be expected, but bone depth varies considerably between individuals.

Characteristic features of trabecular bone in edentulous mandibles.

BACKGROUND: The density and architecture of the alveolar trabecular bone are crucial to the stability of an endosseous implant. A significantly higher implant failure rate can be expected when implants are placed in alveolar bone with reduced density and stability. Therefore, the present study aimed to describe the trabecular bone structure of edentulous mandibles. METHOD: Two hundred and seventy-eight bone sections, including the lateral incisor, first premolar, and first molar regions, were obtained from 128 edentulous lower jaws (68 females and 60 males; mean age: 77.58 years). Ground sections were prepared for each region using the 'sawing and grinding' technique. The following standard structural histomorphometric parameters were determined using a semiautomatic image analysis: trabecular bone volume, trabecular thickness, trabecular number, trabecular separation, and the trabecular bone pattern factor, which describes the connectedness of cancellous bone structures. Also, the maximum height of the jaw section was determined, to detect any possible correlations between vertical height and histomorphometric parameters. RESULTS: All the histomorphometric parameters examined showed an unexpectedly huge range of variation. The mean trabecular bone volume ranged between 20.9% and 36.9%. The mean trabecular thickness showed values between 165.9 and 224.7 microm. The mean trabecular number ranged between 1.22 and 1.77/ mm, and the mean trabecular separation ranged between 436.7 and 720.0 microm. The mean trabecular bone pattern factor showed values between -0.05 and -3.01/ mm. The maximum height of the jaw sections showed values between 16.05 and 23.42 mm. The trabecular bone volume, thickness, number and connectivity were significantly lower in the molar region than in the incisal and premolar regions. Significant sex-specific differences were found in all the regions, female mandibles showing a smaller amount and lower connectivity of cancellous bone than male mandibles. No correlation could be found between the maximum height of the jaw and the histomorphometric parameters of the cancellous bone. CONCLUSION: A possible explanation for the difference in the density between the incisal and the molar region may be that molars are generally lost at an earlier age than anterior and premolar teeth. As a result, atrophy-related resorptive and remodeling processes commence earlier and progress further in this region than in the anterior and premolar regions. Sex-specific differences are probably due to an increased postmenopausal bone loss of the females.

Bone augmentation ability of autogenous bone graft particles with different sizes: a histological and micro-computed tomography study.

OBJECTIVES: The purpose of this study was to investigate the augmentation process and ability of autogenous bone graft particles of two different sizes in a vertical augmentation chamber. MATERIAL AND METHODS: The cranial bones of 24 rabbits were used. Two polytetrafluoroethylene chambers were filled with harvested bone from tibia with small bone (SB; 150-400 microm) and large bone (LB; 1.0-2.0 mm) of the same weight. Animals were sacrificed after 1, 2, 4 and 8 weeks. The samples were analyzed by micro-computed tomography (micro-CT) for quantitative analysis, and embedded in polyester resin as non-decalcified specimens for histological analysis. Total bone volume (TBV), bone height (BH) and distribution of bone structure were calculated by micro-CT. RESULTS: Micro-CT evaluation and histology revealed a significant difference between the investigated specimens. TBV and BH of SB decreased to about 50% of the initial situation, and there was a statistically significant difference between 1 and 8 weeks. In contrast, TBV and BH of LB were almost retained at all experimental time points. Significant differences in TBV and BH were also observed between LB and SB at 8 weeks. Bone volume of SB decreased predominantly in the upper half of the chamber at 4 and 8 weeks. In the histological observations, SB showed favorable new bone formation and rapid bone resorption in a time-dependent manner during the entire experimental period. However, LB exhibited favorable morphological stability and continued new bone formation. CONCLUSION: SB follows a smooth osteogenic process, whereas it is not effective in volume augmentation. LB is superior to SB in augmentation ability.

Determination of bone's mechanical matrix properties by nanoindentation.

Osteoporosis is a devastating disease that is characterized not only by a reduction in bone quantity but also by deterioration in bone quality. The quality of bone tissue is greatly influenced by its mechanical properties and, therefore, investigations into the etiology and enhanced detection of osteoporosis, or the efficacy of interventions, may require the assessment of bone's mechanical properties at the level of the tissue. Nanoindentation is a relatively new technique that is capable of evaluating bone's quasi-static and dynamic mechanical properties on extremely small volumes of tissue. These data can be used directly to describe the pre-yield properties of the matrix, but can also be combined with imaging techniques and mechanical models to extrapolate the mechanical properties from the level of the tissue to that of the organ.

Design of complex bone internal structure using topology optimization with perimeter control.

Large facial bone loss usually requires patient-specific bone implants to restore the structural integrity and functionality that also affects the appearance of each patient. Titanium alloys (e.g., Ti-6Al-4V) are typically used in the interfacial porous coatings between the implant and the surrounding bone to promote stability. There exists a property mismatch between the two that in general leads to complications such as stress-shielding. This biomechanical discrepancy is a hurdle in the design of bone replacements. To alleviate the mismatch, the internal structure of the bone replacements should match that of the bone. Topology optimization has proven to be a good technique for designing bone replacements. However, the complex internal structure of the bone is difficult to mimic using conventional topology optimization methods without additional restrictions. In this work, the complex bone internal structure is recovered using a perimeter control based topology optimization approach. By restricting the solution space by means of the perimeter, the intricate design complexity of bones can be achieved. Three different bone regions with well-known physiological loadings are selected to illustrate the method. Additionally, we found that the target perimeter value and the pattern of the initial distribution play a vital role in obtaining the natural curvatures in the bone internal structures as well as avoiding excessive island patterns.

Moderate hyperhomocysteinemia induced by short-term dietary methionine overload alters bone microarchitecture and collagen features during growth.

AIMS: In general, hyperhomocysteinemia is increasingly appreciated as a risk factor for various diseases, including osteoporosis. However, its effects in non-adults remain largely unknown. Our aim was to determine whether dietary-caused increased homocysteine levels have deleterious effects on bone structure during growth. MAIN METHODS: We developed a model of moderate hyperhomocysteinemia caused by short-term methionine nutritional overload in growing rats. 30-days-old male Wistar albino rats were randomly assigned to either experimental group subject to a 30-days hypermethionine diet or control group. High-resolution 3D assessment of bone geometry and microarchitecture, as well as fluorescence spectroscopic analysis of bone matrix were performed. KEY FINDINGS: Short-term moderate hyperhomocysteinemia (~30µmol/L) achieved in the study notably affected bone and cartilage characteristics. Parameters of the cortical bone geometry in the experimental group indicated peculiar reorganization of the bone cross-section. Trabecular bone microarchitecture was especially sensitive to hyperhomocysteinemia showing clearly negative bone balance in the experimental group (almost 30% reduced bone volume, mainly due to ~25% decrease in trabecular number as well as markedly reduced trabecular connections). Fluorescent spectroscopy of bone matrix revealed multiple alterations to collagen spectra due to homocysteine accumulation in bone, indicative of broken collagenous cross-links. SIGNIFICANCE: Given that appropriate accrual of bone mass during growth has important effects on the risk of osteoporosis in adulthood, understanding the skeletal effects of dietary-induced hyperhomocysteinemia in non-adults is essential for interpreting its importance as a modifiable risk factor for osteoporosis and improving programs to preserve/re-establish bone health.

Trabecular architecture can remain intact for both disuse and overload enhanced resorption characteristics.

The paradigm that bone metabolic processes are controlled by osteocyte signals have been the subject of investigation in many recent studies. One hypothesis is that osteoblast formation is enhanced by these signals, and that osteoclast resorption is enhanced by the lack of them. Reduced, or absent, osteocyte signaling can be an effect of reduced mechanical loading (disuse) or of defects in the canalicular network, due to microcracks. This would mean that bone is resorbed precisely there where it is mostly needed. In our study, we addressed this apparent contradiction. The purpose was to investigate how alternative strain-based local stimuli for osteoclasts to resorb bone would affect remodeling and adaptation of the trabecular architecture. For this purpose, a computer-simulation model was used, which couples morphological and mechanical effects of local bone metabolism to changes in trabecular architecture and density at large. Six resorption characteristics were studied in the model: (I) resorption occurs spatially random, (II) resorption is enhanced or (III) strongly enhanced where there is disuse, (IV) resorption is enhanced or (V) strongly enhanced where there are high strains, i.e. overload, and (VI) resorption is enhanced where there is disuse and where there are high strains. Results showed that the rates of structural adaptation to alternative loading were higher for disuse-controlled resorption than for overload-controlled resorption. Architecture and mass remained stable for all cases except (V) in which the structure deteriorated as in osteoporotic bone. We conclude that, given the potential of osteoblasts to form bone in highly strained areas, based on signals from osteocytes, osteoclast resorption can normally be compensated for.

A Surrogate Measure of Cortical Bone Matrix Density by Long T2 -Suppressed MRI.

Magnetic resonance has the potential to image and quantify two pools of water within bone: free water within the Haversian pore system (transverse relaxation time, T2 > 1 ms), and water hydrogen-bonded to matrix collagen (T2 ∼ 300 to 400 µs). Although total bone water concentration quantified by MRI has been shown to scale with porosity, greater insight into bone matrix density and porosity may be gained by relaxation-based separation of bound and pore water fractions. The objective of this study was to evaluate a recently developed surrogate measurement for matrix density, single adiabatic inversion recovery (SIR) zero echo-time (ZTE) MRI, in human bone. Specimens of tibial cortical bone from 15 donors (aged 27 to 97 years; 8 female and 7 male) were examined at 9.4T field strength using two methods: (1) (1)H ZTE MRI, to capture total (1)H signal, and (2) (1)H SIR-ZTE MRI, to selectively image matrix-associated (1)H signal. Total water, bone matrix, and bone mineral densities were also quantified gravimetrically, and porosity was measured by micro-CT. ZTE apparent total water (1)H concentration was 32.7 ± 3.2 M (range 28.5 to 40.3 M), and was correlated positively with porosity (R(2) = 0.80) and negatively with matrix and mineral densities (R(2) = 0.90 and 0.82, respectively). SIR-ZTE apparent bound water (1)H concentration was 32.9 ± 3.9 M (range 24.4 to 39.8 M), and its correlations were opposite to those of apparent total water: negative with porosity (R(2) = 0.73) and positive with matrix density (R(2) = 0.74) and mineral density (R(2) = 0.72). Porosity was strongly correlated with gravimetric matrix density (R(2) = 0.91, negative) and total water density (R(2) = 0.92, positive). The strong correlations of SIR-ZTE-derived apparent bound water (1)H concentration with ground-truth measurements suggest that this quantitative solid-state MRI method provides a nondestructive surrogate measure of bone matrix density.

Effects of laser vs ultrasound on bone healing after distraction osteogenesis: A histomorphometric analysis.

OBJECTIVE: To assess the effects of low-level laser irradiation vs ultrasound irradiation on bone healing after distraction osteogenesis. MATERIALS AND METHODS: Distraction osteogenesis was performed with rapid maxillary expansion devices (Hyrax-Morelli, Sorocaba, São Paulo Brazil) in 24 rabbits (Oryctolagus cuniculus). After a 2-day latency period, the distraction devices were activated for 10 days at a rate of 1 mm/d. Four groups of six animals were treated as follows: (1) control, (2) laser irradiation on the right side, (3) ultrasound irradiation on the right side, and (4) laser irradiation on the right side and ultrasound on the left side. Histomorphometric analysis was used to assess the bone healing area. Analysis of variance was used to perform the statistical analyses. RESULTS: The influence of low-intensity laser associated with ultrasound irradiation on bone healing was statistically significant. The analyses showed the greatest amount of bone healing in the jaws of animals in group 4, which received treatment with both ultrasound and laser. CONCLUSION: This study concluded that bone healing is accelerated with the application of laser irradiation. The greatest effects were observed with combined ultrasound and laser treatment.

Equivalency of various methods for estimating osteoid seam width.

We compared the true osteoid seam width (TOSW) as measured by a modification of the orthogonal intercept lengths with various methods of estimating seam widths, including (1) the commonly used length measurements at four equidistant points (O.Wi/4PT), (2) osteoid area divided by the osteoid perimeter (O.Ar/O/Pm) or the bone/osteoid interface (O.Ar/B.Bd), and (3) a novel method for estimating seam width defined as the osteoid area divided by the major axis of the seam (O.Ar/Axis). All methods for approximating osteoid seam width significantly exaggerated the true osteoid seam dimension by an amount that ranged from 16 to 23%. However, the relative accuracy of all methods of estimating osteoid seam width are equivalent as evidenced by the similar mean difference from the TOSW (3.4, 4.1, 5.1, and 3.8) demonstrated by O.Wi/5PT, O.Ar/Axis, O.Ar/O.Pm, O.Ar/B.Bd, respectively. Regression analysis of the various estimates of seam width on TOSW also demonstrated the equivalency of these methods. Moreover, all estimates could be employed to discriminate seams of normal dimensions from abnormally wide seams in bone specimens derived from patients with osteomalacia. Differences between the methods, however, were observed that may have practical importance. In this regard, the direct procedure of determining distance demonstrated less variance than the indirect estimate of width. As a result, the direct measurement required fewer samples (n = 13) to detect a significant difference to normal and could discriminate smaller deviations in seam width (1.7 microns) at a given sample size compared with O.Ar/Axis (n = 28; 2.9 microns), O.Ar/O.Pm (n = 42; 3.4 microns), and O.Ar/B.Bd (n = 42; n = 3.2).(ABSTRACT TRUNCATED AT 250 WORDS)

An automated analysis of intracortical porosity in human femoral bone across age.

The matrix of human cortical bone is arranged around a network of vascular spaces (hereafter referred to as "pores"). Our aim was to investigate age-related differences in human cortical porosity (total pore area divided by cortical bone area), pore size and number, and surface to volume ratios, while adjusting for sex, height, and weight. Ninety-six specimens of entire transverse sections of human femoral diaphysis, from subjects aged 21-92 years, were examined. We used our established automated image acquisition and analysis system which measures pores from entire sections of multiple specimens of bone. Over 400,000 pores were recorded. Results showed a greater porosity in older bone (p < 0.01) but marked variation in porosity for any given age. The cohort median, of the specimen medians, of pore cross-sectional area was 2050 microns 2. Older specimens did not have more pores than younger specimens but had a greater proportion of larger pores (p < 0.05) and greater intraspecimen variation in pore size (p < 0.001). The pore surface to bone matrix volume ratio was a median 2.3 mm2/mm3. This varied more than 4-fold between individuals but did not relate to age. No simple relationships were found between any of the measured parameters and either sex, height, or weight, even after adjustment for age. We conclude that the greater porosity in older specimens is due to greater pore size rather than a larger number of pores. Age, however, explains little of the inter-individual variation in the parameters studied.

Cross-link profile of bone collagen correlates with structural organization of trabeculae.

Little is known regarding the mechanisms that govern the structural organization of cancellous bone. In this study, we compare the nature of the collagen in vertebral cancellous bone with the structural organization of its trabecular network. Cylindrical specimens of cancellous bone from vertebrae were obtained from nine autopsy subjects (ages 46-88). In each subject, eight pairs of corresponding samples were obtained from three levels in the spine and three areas within the vertebral body, leading to a total of 68 pairs of samples. The cylinders from one side were used for morphometry and the classical morphometrical parameters were obtained (BV/TV, bone volume fraction; Tb.Th, trabecular thickness; Tb.N, number; Tb.Sp, trabecular spacing) and strut analysis (TSL, total strut length; Nd, number of nodes; Fe, number of free-ends). The amount of osteoid bone was also quantified. The cylinders from the other side were powdered and used for collagen assessment, including the amount of collagen (% w/w), and its content in immature cross-links; such as hydroxylysinonorleucine (mol/mol of collagen) and dihydroxylysinornorleucine, as well as stable mature cross-links, such as hydroxylysylpyridinoline (HP), lysylpyridinoline (LP), and the pyrrole cross-links. A random regression model was used to explore the correlations. None of the biochemical parameters correlated with the BV/TV except the ratio between immature and mature cross-links (eta(2) = 0.34, p < 0.05). There was no relationship between the amount of osteoid bone and the cross-link profile. However, the concentration of pyrrole and HP cross-links in the bone samples correlated with the structural organization of its trabeculae, but in an opposite direction. Hence, the pyrrole/HP ratio was a good predictor of Tb.Th, Tb.N, Tb.Sp, and TSL (eta(2) > 0.65 and p < 0.01) as well as Fe and star marrow space (eta(2) > 0.45 and p < 0.05). The cylinders from subjects with high pyrrole or low HP in their bone collagen had a relatively thick and simple structure. Those with low pyrrole and high HP had relatively thin trabeculae that were more numerous and spread over a complex network. The relative concentrations of the pyrrole and pyridinoline cross-links appear to reflect the structural organization of the trabeculae.

The effect of demineralized bone matrix on the healing of intramembranous bone grafts in rabbit skull defects.

A clinical dilemma exists regarding the type of bone that should be used to replace diseased or traumatized osseous tissue. Oral, plastic, and orthopedic surgeons normally implant viable mineralized endochondral (EC) autografts or demineralized EC allografts. A few clinicians have recognized the disadvantages of using EC bone in craniofacial surgery and advocated the replacement of intramembranous (IM) bone with healthy IM bone. However, controversy and uncertainty surround our understanding of these matrices to induce bone formation. Recent studies have advocated the use of other materials with osteoinductive properties, such as demineralized bone matrix (DBM). The proposed delivery system used in this study included IM bone grafts, DBM, and fixation of the IM bone graft. The purpose of this work was to gain further insights into the mechanism of healing of IM bone, in both the presence and the absence of DBM, and to compare the healing of IM bone grafts with that of DBM alone. Critical-sized (10 x 5 mm), full-thickness bony defects in rabbit parietal bone, devoid of periosteum, were filled with IM bone graft (mandible) alone, demineralized cortical bone matrix (DBM) alone, or combined DBM-IM bone graft, or were left unfilled. Histologic changes were examined 14 days later. The IM bone graft healed through IM ossification with no intermediate cartilage stage. DBM and composite DBM-IM healed through an EC ossification with an intermediate cartilage stage. It is hypothesized that the role of the IM graft is to induce neovascularization into the defect site, and that the undifferentiated mesenchymal cells in the perivascular region of the new blood vessels are induced by the bone morphogenetic protein(s) in the DBM into bone-forming cells.

Over expression of bone morphogenetic protein-3b (BMP-3b) using an adenoviral vector promote the osteoblastic differentiation in C2C12 cells and augment the bone formation induced by bone morphogenetic protein-2 (BMP-2) in rats.

BMP-3b is a novel BMP-3-related protein and its biological functions are unknown. In order to investigate the biological actions of BMP-3b, we constructed a BMP-3b-expressing recombinant adenoviral vector (AxCAKBMP-3b). We show that over expression of BMP-3b stimulated the induction of differentiation and the osteoinduction activity of a human BMP-2-expressing recombinant adenoviral vector (AxCAOBMP-2). C2C12 cells were infected in vitro with AxCAKBMP-3b, AxCAOBMP-2 and a control vector containing no foreign genes (AxCAwt). Cells infected with AxCAOBMP-2 and AxCAKBMP-3b produced more alkaline phosphatase and secreted more osteocalcin into the culture medium than cells infected with AxCAOBMP-2 and AxCAwt. When AxCAOBMP-2, AxCAKBMP-3b, and AxCAwt were injected into the calf muscles of nude rats (F 344/N Jcl-rnu), the osteoinduction seen with AxCAOBMP-2 and AxCAKBMP-3b was greater than that seen with AxCAOBMP-2 and AxCAwt.

Measurements of trabecular bone structure in man.

A quantitative analysis of trabecular bone structure is presented, based on omnidirectional distributions of paths across (a) trabeculation and (b) marrow cavities. The omnidirectional distributions, which take into account structural anisotropy, are generated from measured distributions of paths. Representative examples are given, together with values of two commonly quoted structural parameters, the ratio of endosteal surface to bone volume and percentage bone volume. Data on the biological and age variations in the third lumbar vertebra are also presented and an index of trabecular anisotropy suggested. Finally, the results are compared with those of other workers and estimates of useful skeletal parameters given.