Optimization of corticosteroid induced osteoporosis in ovariectomized sheep. A bone histomorphometric study.

During osteoporosis induction in sheep, side effects of the steroids were observed in previous studies. The aim of this study was to improve the induction regimen consisting of ovariectomy, calcium/vitamin D- restricted diet and methylprednisolone (-MP)- medication with respect to the bone metabolism and to reduce the adverse side effects. Thirty-six ewes (age 6.5 +/- 0.6 years) were divided into four MP-administration groups (n = 9) with a total dose of 1800 mg MP: group 1: 20 mg/day, group 2: 60 mg/every third day, group 3: 3 x 500 mg and 1 x 300 mg at intervals of three weeks, group 4: weekly administration, starting at 70 mg and weekly reduction by 10 mg. After double-labelling with Calcein Green and Xylenol Orange, bone biopsy specimens were taken from the iliac crest (IC) at the beginning and four weeks after the last MP injection, and additionally from the vertebral body (VB) at the end of the experiment. Bone samples were processed into stained and fluorescent sections, static and dynamic measurements were performed. There were no significant differences for static parameters between the groups initially. The bone perimeter and the bone area values were significantly higher in the VB than in the IC (Pm: 26%, p < 0.0001, Ar: 11%, p < 0.0166). A significant decrease (20%) of the bone area was observed after corticosteroid-induced osteoporosis (p < 0.0004). For the dynamic parameters, no significant difference between the groups was found. Presence of Calcein Green and Xylenol Orange labels were noted in 50% of the biopsies in the IC, 100% in the VB. Group 3 showed the lowest prevalence of adverse side effects. The bone metabolism changes were observed in all four groups, and the VB bone metabolism was higher when compared to the IC. In conclusion, when using equal amounts of steroids adverse side effects can be reduced by decreasing the number of administrations without reducing the effect regarding corticosteroid-induced osteoporosis. This information is useful to reduce the discomfort of the animals in this sheep model of corticosteroid-induced osteoporosis.

Intra-articular calcium phosphate cement: Its fate and impact on joint tissues in a rabbit model.

Clinical application of injectable ceramic cement in comminuted fractures revealed penetration of the viscous paste into the joint space. Not much is known on the fate of this cement and its influence on articular tissues. The purpose of this experimental study was to assess these unknown alterations of joint tissues after intra-articular injection of cement in a rabbit knee. Observation periods were from 1 week up to 24 months, with three rabbits per group. Norian SRS cement was injected into one knee joint, the contralateral side receiving the same volume of Ringers' solution. Light microscopic evaluation of histologic sections was performed, investigating the appearance of the cement, inflammatory reactions, and degenerative changes of the articular surface. No signs of pronounced acute or chronic inflammation were visible. The injected cement was mainly found as a single particle, anterior to the cruciate ligaments. It became surrounded by synovial tissues within 4 weeks and showed signs of superficial resorption. In some specimens, bone formation was seen around the cement. Degeneration of the articular surface showed no differences between experimental and control side, and no changes over time became apparent. No major degenerative changes were induced by the injected cement. The prolonged presence of cement still seems to make it advisable to remove radiologically visible amounts from the joint space.

Mineralization and mechanical properties of the canine mandible distraction wound following acute molding.

To investigate the influence of an acute single step callus manipulation immediately after distraction on mechanical properties and mineralization of the regenerate, custom made distraction devices were bilaterally placed in the mandibular angle of 15 beagle dogs, allowing to simultaneously compress and stretch the regenerate after completed linear distraction. The animals were divided in three groups (n=5): Group 1 and 2 underwent manipulation of the regenerate, group 3 remained in a linear position. After 42 (group1) and 90 (group 2 and 3) days of consolidation the animals were sacrificed. The mechanical properties were assessed in an Instron testframe and bone density quantified by quantitative computed tomography and three- dimensionally assessed (Scion Image processing and analysis software). After 6 weeks of consolidation 25% of the specimens reached a stiffness which was >/=90% of the mean values of the unoperated reference hemi-mandibles. After a 13 week consolidation period, 62.5% were as stiff as the referenced specimens. Manipulated regenerates, allowed to heal under stable conditions for 13 weeks, had the same mechanical properties as specimens that underwent pure linear distraction. A temporary but not significant delay of osseous healing had to be postulated for the stretched zone after 6 weeks, indicating this area to be more critical than the compressed area.

Histology of the porcine mandibular distraction wound.

The purpose of this study was to document the progression and pattern of endosteal bone formation in a porcine mandibular distraction wound. Bone formation was assessed in a 0-day latency model (n=24 pigs) using distraction rates of 1, 2, or 4 mm/day to create a 12-mm gap. Macro-radiographs and sagittal histologic sections, from the center of the mandible, were evaluated by computer morphometrics (% bone fill) and by a semi-quantitative bone formation score. Mean percent area of new bone was 12.4% (0-25.9%), 7.5% (0-21.3%) and 3.8% (0-10.5%) in mandibles distracted at 1, 2 or 4 mm/day respectively. At all time points, percent area of new bone was highest in mandibles distracted at 1 mm/day. Bone was deposited from the margins of the osteotomy toward the center of the wound and occurred first around the inferior alveolar canal and tooth bud regions. New bone formed by intramembranous ossification alone. The results of this experiment document the contribution of endosteal bone formation in this model of distraction osteogenesis.

Effect of bone size, not density, on the stiffness of the proximal part of normal and osteoporotic human femora.

Proximal femur fractures in the elderly lead to a high rate of hospitalization. In studying the operative treatment of such fractures, it is first necessary to understand the relationship between the morphologic properties of this part of the femur (related to both geometry and density) and its mechanical properties. Numerous investigations showed that femoral strength correlates with bone mass; however, the dimension of the bones was not taken into account. We measured the relationship between the stiffness of the proximal femur under physiologic load and its geometry and density. Conventional x-rays and quantitative computed tomography (QCT) were carried out on pairs of human cadaver femora. Eight pairs were instrumented with strain gauges. Bones were subjected to an eccentric load that simulates moderate weight bearing (single-leg stance), and the strain parallel to the bone axis was plotted as a function of the load applied. An apparent bone stiffness was calculated as the ratio between the strain gradient within the section and the load applied. Strong correlation was found between x-ray densitometry and QCT. The bone stiffness also correlates strongly with the geometry (area) and slightly with bone mass; however, an unexpectedly low correlation was found between stiffness and density. We chose stiffness as a mechanical parameter (not strength) because it describes the "normal" bone behavior under load. Our results clearly demonstrate that the cross-sectional size of the bones must be taken into account when establishing the relationship between the mechanical characteristics of the bone and its morphology. In accordance with previous predictions, our results support that bone loss due to osteoporosis is compensated for by an increase in bone diameter.

Long-term in vivo degradation and bone reaction to various polylactides. 1. One-year results.

Injection-moulded pins from poly(L-lactide), poly(L/DL-lactide) (95/5%) were implanted in the cortex of the tibiae of sheep. The bone-implant interface was evaluated to observe whether there is any bone resorption caused by the implants. The molecular weight and crystallinity changes upon implantation were also measured. There was no net bone loss around the implants or sterile cyst formation in any of the animals implanted with polylactides up to 1 year. The new bone formed around the poly(L-lactide) and poly(L/D-lactide) pins was separated from the implants with a thin layer of connective tissue. For the implants from poly(L/DL-lactide), there was direct apposition of bone on the polymeric material. At 1 year of implantation, the implants were not completely resorbed, although the molecular weight of polylactides was reduced from 40,000-50,000 to 500-300. The crystallinity at 1 year was about 45% for poly(L/DL-lactide) and poly(L/DL-lactide) and 65% for poly(L-lactide), respectively, indicating the presence in the degraded material of thermodynamically stable crystals.

Biomechanics of the craniocervical region: the alar and transverse ligaments.

In the treatment of spine fractures and fracture-dislocations, stability of the spine is one of the major objectives. In the craniocervical joint, the alar and transverse ligaments provide much of the stability of the healthy spine. Because the anatomy appears well described, the contribution of each of these structures so far has received little attention. The alar ligament restrains rotation of the upper cervical spine, whereas the transverse ligament restricts flexion as well as anterior displacement of the atlas. A lesion in one or both structures can produce damage to the neural structures and/or cause pain. To investigate the possible role of each of these ligaments, a mechanical and histologic study of the upper cervical spine was made. The bone-ligament-bone complex of the alar and transverse ligaments was subjected to uniaxial mechanical testing in seven specimens. The alar ligaments had an in vitro strength of 200 N, and the transverse ligaments had an in vitro strength of 350 N. Histologic analysis revealed a mainly collagenous nature of these ligaments. Clinical evidence (broken odontoid processes) suggests that the transverse ligament is strong enough to withstand physiologic loads. The alar ligament, on the other hand, due to its lower strength and its axial direction of loading, might be prone to injury and therefore require stabilization of the appropriate vertebra more often than normally is assumed.

Juxtaarticular bone loss in experimental inflammatory arthritis.

The osteopenia associated with experimental inflammatory arthritis was studied by a histomorphometric method that exphasized net changes in bone composition. Juxtaarticular trabecular bone volume and turnover were studied in the carrageenan injection model of inflammatory arthritis of the mature rabbit knee. Trabecular bone volume was studied by histomorphometry of the femoral condyles and confirmed by photodensitometry on standard macroradiographs. Osteogenesis was studied by imaging of calcein fluorochrome-labeled newly formed bone in undecalcified histological sections. A significant net loss of cancellous bone (approximately 20%) occurred over 49 days in both the medial and the lateral femoral condyle in the arthritis group compared with normal controls. Total osteogenesis was increased fourfold and it was calculated that an even greater increase in total bone resorption was responsible for the negative bone balance. There is evidence that the periarticular bone loss of human rheumatoid arthritis is also associated with increased bone turnover. Quantitative studies of the kinetics of bone remodeling in inflammatory arthritis will provide the basis for therapeutic attempts to prevent or reverse arthritis-induced bone loss. Fracture risk in inflammatory arthritis may be increased not only by osteopenia, but additionally by the presence of a large proportion of newly formed (and presumably less mineralized) bone.

Healing of cancellous bone osteotomy in rabbits--Part I: Regulation of bone volume and the regional acceleratory phenomenon in normal bone.

We performed a simple, undisplaced, incomplete osteotomy of the distal medial condyle of mature New Zealand White rabbits and permitted healing without fixation for 4 weeks. During the recovery period, the rabbits received the bone label calcein in their drinking water. From ground stained and unstained sections, we determined histomorphometric parameters in five zones: osteotomy gap, juxtaosteotomy, lateral condyle, metaphysis, and diaphysis. At 4 weeks, newly formed cancellous bone had obliterated the osteotomy gap. Osteotomy also stimulated new bone formation in cancellous bone of the medial condyle around the osteotomy. At that site, we found a fivefold increase in new bone without a change in bone volume (BV). Stimulation of bone formation was not observed in cancellous bone of the lateral condyle that had not been operated on or in cortical bone of the metaphysis or diaphysis. The maintenance of BV, despite increased new bone formation in the osteotomized medial femoral condyle, was probably due to a local acceleration of bone remodeling.

Healing of cancellous bone osteotomy in rabbits--Part II: Local reversal of arthritis-induced osteopenia after osteotomy.

With use of the intravital bone label calcein, we previously identified, by histomorphometry, greatly increased new bone formation and juxtaarticular osteopenia (a high turnover state) in carrageenan-induced experimental inflammatory arthritis of the knee in rabbits. In Part I of this paper, we showed that a large increase in new bone formation after a stable incomplete distal femoral condylar osteotomy of normal bone in the rabbit does not increase bone volume (BV) in normal tissue adjacent to the osteotomy (zone 1). In this study, we performed an osteotomy of bone made osteopenic by experimental inflammatory arthritis. After 4 weeks of healing, we examined new bone formation and BV in five zones around the osteotomy. In zone 0 (the osteotomy gap), new bone formation filled the gap to normal BV, as we found for the normal/osteotomy group. In zone 1 (adjacent to but excluding the osteotomy gap), we found greatly increased new bone formation and a return to normal BV, increased from the osteopenic level induced by experimental inflammatory arthritis. In this model, osteotomy apparently reversed the bone-wasting process of inflammatory arthritis. This effect was not observed in other zones: the lateral condyle that had not been operated on, the metaphysis, or the diaphysis.

Laser Doppler flowmetry for bone blood flow measurement: correlation with microsphere estimates and evaluation of the effect of intracapsular pressure on femoral head blood flow.

Laser Doppler flowmetry (LDF) was used to measure bone blood flow in the rabbit femoral head and femoral condyles. To correlate the LDF output signal blood cell flux to in vivo blood flow, simultaneous measurements using LDF and 85Sr-labeled microspheres were made in an adult rabbit model. There was no correlation between the two methods for blood flow in the femoral condyles and the correlation between the two methods for blood flow in the femoral head does not achieve statistical significance. An LDF signal of 0.4 V was approximately equal to a microsphere measured flow rate of 0.4 ml blood/g bone/min. The strength of the correlation in the latter case may have been affected by (a) large arteriovenous shunts, (b) inadequate mixing of the microspheres with a left ventricular injection, and (c) insufficient numbers of microspheres present in the bone samples with which to satisfy the mathematical requirements of the microsphere method. When LDF was used to evaluate the effect of elevated intracapsular pressure on femoral head blood flow in skeletally mature rabbits, femoral head subchondral bone blood flow declined with increasing intracapsular pressure from a baseline value of 0.343 +/- 0.036 to a value of 0.127 +/- 0.27 at 120 cm of water pressure. The decline in femoral head blood flow was statistically significant at pressures of 40 cm of water or higher (p less than 0.001), and evaluation of sections of the proximal femora made from preterminal disulphine blue injections confirmed these findings. Intracapsular tamponade has an adverse effect on femoral head blood flow beginning well below central venous pressure and should be considered in the pathophysiology of posttraumatic and nontraumatic necrosis of the femoral head. Laser Doppler flowmetry was easy to use and appears to be a reproducible technique for evaluating femoral head blood flow, offering distinct advantages over the microsphere technique for measuring bone blood flow. Further studies of the in vivo calibration of the LDF method for bone blood flow are necessary for the method to have potential for clinical application.

Resorption patterns of calcium-phosphate cements in bone.

Two calcium phosphate cements, one monophasic and the other biphasic, have been used as bone void filler in a sheep model. The cements were injected into a slot defect in the proximal tibia and into a cylindrical defect in the distal femur. In this study, we focused on the resorption pattern of the two cement formulations and the subsequent biologic reaction. Bone remodeling occurred synchronously with the resorption of the implant material in a creeping substitution process. Cracks and pores in the monophasic cement were filled with osseous tissues. The biphasic cement showed faster resorption of the matrix. The more slowly resorbing granules were surrounded by newly grown bone, thus providing an inverse scaffold for cancellous bone regeneration. In highly loaded areas, the long-term support function of the fixation appears to be critical. Because cortical bridging of the defects was seen in only one case, it can be concluded that calcium-phosphate cements are preferentially suitable as cancellous bone substitute materials.

Histology of the alar and transverse ligaments.

Seven sets of human transverse and alar ligaments, after tensile testing, and eight corresponding ligaments without testing, were examined histologically with respect to their fiber composition and fiber orientation. Various staining techniques were supplemented by polarized light microscopy. Both the transverse and the alar ligaments consist of collagen fibers, with very few elastic fibers in the peripheral layer. In the central portion of the transverse ligament, the collagen fibers cross each other at an angle of approximately 30 degrees. Close to the dens, the transverse ligaments show on their ventral side a transition into fibrocartilage. Except for the immediate site of failure, no differences became evident between tested specimens and controls. The collagen, as the almost exclusive constituent, together with the fiber orientation determine the mechanical properties of these ligaments. This supports the hypothesis that the ligaments could be irreversibly overstretched or even ruptured when the head is rotated and, in addition, flexed by impact trauma, especially in unexpected rear-end collisions.

A paradigm of delayed union and nonunion in the lumbosacral joint. A study of motion and bone grafting of the lumbosacral spine in sheep.

In a group of nine sheep (Group A), it was noted that when small, interlocking cancellous and cortical cancellous bone grafts are placed posteriorly on the lumbosacral spine, union always occurred in the interlumbar motion segments and almost never occurred at the lumbosacral joint. One of the main differences in these two areas is the amount of motion that occurs at each level with flexion and extension. Because nonunion following bone grafting for arthrodesis of the spine is a serious clinical problem, we have studied the amount of motion seen at the interlumbar and lumbosacral joints in sheep to ascertain how much motion is compatible with union and how much is associated with nonunion. In vivo studies were carried out in eight sheep (Group B), and five normal spine segments were studied in vitro to determine normal motion in this species. To simplify the complex in vivo motion that occurs at the lumbar motion segments, the simple linear displacement and strain of the fusion mass (consisting of fibrous tissue and bone grafts placed on and between the laminas posteriorly) was measured with the spine in flexion and extension. When the displacement and strain at the interlaminar level of the L6-S1 joint was measured, the linear displacement was found to be 5.2 mm and the associated linear strain 36%. The displacement at the L5-L6 interspace was 1.2 mm, and the strain 10%. The stiffness of the L5-L6 joint (which always fused) and the L6-S1 joint (which did not fuse, with one exception) were also studied. In a third group of four animals (Group C), internal fixation of the lumbosacral joint was attempted in addition to bone grafting.(ABSTRACT TRUNCATED AT 250 WORDS)

Bone growth and remodelling after fracture.

We used a rabbit model to investigate the mechanism by which the angulation of fractures is corrected in children. We produced a transverse proximal tibial fracture in one leg of 12 eight-week-old New Zealand white rabbits and measured bone alignment and length and the patterns of bone growth and remodelling. The angle between the joint surfaces changed rapidly to correct the alignment of the limb as a result of asymmetrical growth of epiphyseal plates. In an adult with closed plates, the angle between the joint surfaces cannot therefore improve. The angle at the fracture itself showed slow improvement because of bone drift and the asymmetrical growth of the epiphyseal plates. Remodelling corrected the shape of the bone in the region of the fracture. Periosteal division on the convex side increased the growth of the epiphyseal plate on that side, thus slowing the correction. The effect was relatively small, providing an indication that factors other than the periosteum are important in inducing correction. External torsional deformities developed because of helical growth at the plate. This was probably caused by abnormal posture which induced a torque at the growth plate. Helical growth is the mechanism by which rotational deformities can occur and correct.

Theoretical considerations in rigid fixation of facial bones.

The classic facial fracture patterns have gradually been changing toward more severe comminutions. This necessitates an early and accurate anatomical reduction and a fixation in this position until the fragments have united. Stable fixation favors early mineralization in the fracture gap. The mechanical function of the implants is thus only needed for a short term. Similar considerations are true for the incorporation of grafts. Mechanics of fixation of facial bones is influenced by the structural properties as well.

Further development of titanium miniplate fixation for mandibular fractures. Experience gained and questions raised from a prospective clinical pilot study with 2.0 mm fixation plates.

Miniplate systems are often used instead of more rigid systems for the treatment of mandibular fractures. While the most stable fixation method for all mandibular fractures is the 2.7 mm plate, most fracture sites and types are eminently suitable for miniplate fixation via an intraoral approach. However, the relatively low stability of the miniplate systems compared with rigid plate systems limits the indications for their use in mandibular fracture treatment, especially when immediate postoperative function is desired. A more rigid miniplate which provides increased stability was studied. The results of a preliminary study and a clinical trial of a 2.0 mm titanium miniplate system are presented in this paper. The therapeutic consequences of the lesser stability afforded by small plate systems are discussed. Indications for miniplate fixation without additional immobilization are reviewed.

Fixation effects on membranous and endochondral onlay bone-graft resorption.

Difficulties arise in the prediction of maintenance of graft volume over time when bone grafts are used for facial contour reconstruction. We hypothesize that graft fixation will decrease movement and lead to decreased resorption. Fixed and nonfixed endochondral (rib) and membranous (skull) onlay bone grafts measuring 30 X 10 X 4 mm were grafted to the mandible bilaterally in 10 adult sheep. Fixation was achieved using the lag-screw technique. Volume measurements using caliper technique were made 20 weeks postoperatively. The volume of graft present at 20 weeks was significantly greater for the fixed bone grafts (p less than 0.001): fixed membranous, 85.9 percent; fixed endochondral, 76.2 percent; nonfixed membranous, 55 percent; and nonfixed endochondral, 16.6 percent. The results are explained using biomechanical theories related to the effects of strain. At present, it is suggested by this study that when onlay bone grafts are stabilized, improved results with respect to graft resorption can be expected.

Fixation effects on membranous and endochondral onlay bone graft revascularization and bone deposition.

This paper investigates the relationship between bone resorption, the process of bone revascularization, and graft fixation. Vital staining techniques and microangiography were used to study the extent of graft revascularization of fixed and nonfixed endochondral (rib) and membranous (skull) onlay bone grafts in 20 adult sheep mandibles bilaterally. This assessment was carried out at 2 and 20 weeks postoperatively. Sequential fluorochrome staining was performed to examine the pattern of new bone deposition. Fixation was achieved using the lagscrew technique. At 2 weeks, membranous bone demonstrated a greater area of graft revascularization if fixed than if the graft was not fixed. The opposite result was seen for endochondral grafts, where nonfixed grafts showed a greater area of revascularization than fixed grafts. At 20 weeks, all bone that was present was fully vascularized. The inconsistencies in the results on the relationship between fixation and revascularization for membranous and endochondral grafts in the early stages of healing (2 weeks) suggest that although revascularization is a necessary precondition for bone resorption and deposition, biomechanical and structural factors may be a more satisfactory explanation for the differences observed in the maintenance of bony volume.

Comparison of compression and torque measurements of self-tapping and pretapped screws.

The choice of an internal fixation system for maxillofacial surgery is made difficult because of lack of information with respect to functional load. This study attempted to clarify some of the controversy with respect to maxillofacial use of these implants. Maximal compressive force to torque values were measured in standardized bone thicknesses of 1, 2, 3, and 4 mm. The screws tested were pretapped AO 1.5-, 2.0-, 2.7-, and 3.5-mm rescue screws and self-tapping Luhr, Champy, and AO 1.5- and 2.0-mm screws. Ten measurements were made for each screw type/bone thickness combination using a piezoelectric washer and torque screwdriver. It was apparent that for 1- and 2-mm bone thicknesses the use of self-tapping screws resulted in the highest compression values. In 3- and 4-mm bone thicknesses, pretapped screws offered the highest compression values. As expected, self-tapping screws had the highest torque values on insertion owing to torque loss in cutting the screw threads. The 2.7-mm screw offered no advantage over the 2.0-mm screws in 1- and 2-mm bone thicknesses but resulted in higher compression values in 3- and 4-mm bone thicknesses.