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Behavior of cortical bone grafts under different types of fixation.

The fate of cortical bone grafts, rigidly fixed by means of a special metal splint, was studied in rabbits. Cortical grafts of fresh autologous bone were compared with dry-stored allogenous transplants frozen to -20 degrees C. To study the incorporation and remodeling, fluorochrome labeling and microradiography were used. The main parameters of remodeling were the quantities of old lamellar bone, newly formed lamellar bone, and vascular spaces present in the transplant areas. The data obtained are compared with the results of a previous similar study of grafts, which were fixed by Mersilene sutures. Fresh autologous as well as allogenous (dry-stored, frozen to -20 degrees C) cortical bone grafts, used as free transplants, show the same type of incorporation and ultimately the same amount of remodeling. Rigid fixation is a valuable contribution in promoting primary function.

Animals↗

Vascular changes in replanted and autotransplanted apicoectomized mature teeth of dogs.

The purpose of the present investigation was to study whether international fracturing of the root tip at the time of replantation or autotransplantation of mature teeth might improve the revascularization capacity of the pulp. The vascular system in apicoectomized and nonapicoectomized replanted and autotransplanted mature dog teeth was studied 10, 30 and 120 days postoperatively by means of a barium sulfate injection medium combined with microradiography. Pulpal revascularization occurred during a 120-day period in eight out of nine apicoectomized teeth. The pulp of the nonapicoectomized teeth did not revascularize.

Angiography↗

Impaired tensile strength after shock-wave application in an animal model of tendon calcification.

Extracorporeal shock-wave application facilitates dissolution of rotator cuff calcifications. Therefore, disappearance or disintegration of tendon calcifications by shock waves might be appropriate for any kind of tendon calcification. Here, shock waves with various energy flux densities were applied to the mineralized medial gastrocnemius tendon of turkeys as an animal model. After application of shock waves in vivo, with energy flux density of 0.6 mJ/mm(2), histologic examination and microradiography did not show dissolution or disintegration of tendon calcifications. After shock-wave application in vitro, even for energy flux density of 1.2 mJ/mm(2) neither dissolution nor disintegration of tendon calcifications were observed. Biomechanical testing revealed significant impairment of tensile strength following shock-wave application in vitro, with energy flux density of 1.2 mJ/mm(2), but not with 0.6 mJ/mm(2). These results are important for considerations of clinical extracorporeal shock-wave application on tendon calcifications, as well as on tendon ossifications.

Animals↗

Digital venous anatomy.

The veins of the digits and their valves were studied in nine cadaveric hands by sequential angiography, microdissection, routine histology, cross-sectional microradiography, and corrosion casting. The findings revealed a pattern of dorsal venous arches situated over each digit. Connections between those arches were at the level of the metacarpal heads, the point at which most of the palmar venous blood joins the dorsal system by intercapitular veins. A system of valves, which were arranged so as to direct flow from distal to proximal, from palmar to dorsal, and from radial to ulnar in the hand, was present in all veins as far distal as the pulp.

Fingers↗

Recombinant human bone morphogenetic protein-4 (rhBMP-4) enhanced posterior spinal fusion without decortication.

In posterior spinal fusion, insufficient decortication may decrease the number of bone marrow derived ostoprogenitor stem cells and affect the success of bony fusion. The finding of bone formation through interaction between rhBMP-4 and non-marrow derived mesenchymal cells constituted the basis of the current study. The aim is to investigate the possibility of molecular enhancement of posterior spinal fusion by site-specific application of rhBMP-4 with or without surgical decortication. Eighteen adult rabbits underwent single level bilateral posterior intertransverse process spinal fusion at L5-L6. one side with decortication, and the other side without decortication. Two animals underwent sham operation without bone grafts, the other 16 animals were randomly allocated into three groups, using hydroxyapatite-tricalcium phosphate (HA-TCP) ceramic blocks augmented with 0, 125 and 5 micromg [corrected] of rhBMP-4 respectively. Spinal fusion morphology was evaluated with sequential X-ray, microradiography and histomorphology. At week 7, complete bony fusion was achieved in none of the groups without rhBMP-4 irrespective of whether the bony contact surface was decorticated or not. In the groups with low dose rhBMP-4, complete fusion occurred in two of six un-decorticated sites (33%) and in three of six (50%) decorticated sites. 100% complete fusion was found in the high dose rhBMP-4 group independent of surgical decortication. The dorsal cortices of the un-decorticated transverse processes were replaced by newly formed trabecular bone through biological remodeling. This study suggested that rhBMP-4 can induce non-marrow derived mesenchymal cells to differentiate into osteogenic cells and thus enhance the high success rate of pesterior spinal fusion in both the decorticated and un-decorticated model.

Animals↗

Autologous osteoblasts enhance osseointegration of porous titanium implants.

The goal of this study was to assess the osseointegration of porous titanium implants by means of coating with autologous osteoblasts. Titanium implants (8 x 5 x 4 mm) having drill channels with diameters of 400, 500, and 600 microm were coated with autologous osteoblasts obtained from spongiosa chips. The implants were inserted into the distal femora of 17 adult Chinchilla Bastard rabbits (group I). Uncoated implants were inserted as controls in the contralateral femur (group II). The animals were sacrificed after 5, 11, and 42 days. Intravital fluorochrome labeling and microradiography were used for the assessment of bone ingrowth into the titanium channels. In both groups, no bone tissue was formed in the channels up to day 5. On day 11, group I exhibited significantly more (p<0.05) bone tissue (19.8+/-14.0% vs. 5.8+/-9.1%) with greater bone-implant contact (13.3+/-15.1% vs. 5.7+/-5.3%, p<0.05) at the channel mouths than group II. Bone tissue was formed mainly between day 15 and 30 in group I, in group II between day 25 and 40. Six weeks after implantation, bone tissue filled on an average 68.8+/-15.1% of the mouths of the drill channels in implants in group I, the filling for group II was 49.8+/-18.1% (p<0.05). The average bone-implant contact at the channel mouths after six weeks was 56.5+/-13.5% in group I, 40.2+/-21.9% in group II (p<0.05). 600-microm channels showed at this time point the best osseous integration (p<0.05). Coating with autologous osteoblasts accelerates and enhances the osseointegration of titanium implants and could be a successful biotechnology for future clinical applications.

Animals↗

Osteopontin is associated with bioprosthetic heart valve calcification in humans.

Calcification of non-osseous tissues such as heart valves or vessels is a major concern in clinical practice. The exact mechanism is still unknown. Numerous studies have shown that mineral deposits of crystalline hydroxyapatite within these tissues were associated with increased non-collagenous protein content. More recently osteopontin was found to be associated with calcification in living tissues such as vessels and native human aortic valves. The aim of this study was to determine whether or not non-collagenous proteins can also be found in non-living tissues such as glutaraldehyde-pretreated porcine valves after implantation in humans. Thirty-eight glutaraldehyde pretreated porcine bioprostheses were studied: 16 not implanted and 22 after 11 years of implantation in the aortic and mitral valve position in humans. In areas of calcification vizualized by Von Kossa staining and microradiography, immunostaining using polyclonal antibodies against calcium-binding proteins showed osteopontin positive staining and no staining for osteocalcin, bone sialoprotein or osteonectin. In uncalcified areas and in non-implanted values, staining for osteopontin or other calcium-binding proteins was negative. Western blot analysis of macroscopically calcified and uncalcified areas showed that several proteins were adsorbed in implanted values and confirmed the presence of osteopontin in the calcified areas, while no immunolabelling was found in non-calcified areas, in uncalcified valves and in non-implanted valves. Thus the presence of osteopontin in the calcified areas of bioprosthetic heart valves implanted in human indicates that this protein is associated with bioprosthetic valvular calcification. Since these values are made of non-living connective tissue, and no cell immunostained for osteopontin was found around the calcified area, this suggests that a non-cellular mediated mechanism involving protein adsorption may play a role in bioprosthetic valvular calcification.

Bioprosthesis↗

The effect of hydroxyapatite coating on ingrowth of bone into cavities in an implant.

A canine implant study was performed to assess whether hydroxyapatite (HA) could induce bone to close gaps at the bone-prosthesis junction. Titanium alloy (TiAlV) plates with varying grooves cut into them (1, 2, and 3 mm deep) and identical on both sides were sprayed on one side with HA and left as bare metal on the other. These were then implanted into the distal femora of dogs that were killed 4 and 8 weeks afterwards. Specimens were analyzed using optical microscopy and microradiography. Results show that the presence of HA induces bone to grow almost completely down the deepest groove by 8 weeks. In the 1- and 2-mm grooves at 4 weeks the depth of penetration of bone into the grooves was greater on the HA coated side (but the actual volume of bone in the grooves was greater on the TiAlV side because the bone present was more dense). We conclude that the presence of HA induces bone to close relatively large gaps. Although bone does not grow all the way into uncoated defects there can be more bone here in the short term. This feature of HA may be of great value in hip implant fixation.

Animals↗

Quantitative measurements of remineralization of incipient caries.

White demineralized areas after the removal of orthodontic appliances remain a problem for clinicians and patients. The main objective of this study is to assess clinical photography as a method for the evaluation of caries and white spots and the in vivo study of remineralization of carious lesions. The secondary objective was to compare the effect of different levels of both experimental and commercial fluoride solutions on the remineralization of enamel carious lesions. White spot lesions were created with an acid solution on extracted permanent human teeth, and the lesions were remineralized in remineralizing solutions with or without fluoride ions. The changes in the enamel surface during the demineralization and the remineralization processes were recorded with a 35 mm clinical camera. Photographs were taken of the experimentally created white spots, the samples were sectioned for microradiography testing, and the actual mineral contents of the white spot lesions were calculated from the microradiographs. The enamel lesions were further analyzed by powder x-ray diffraction to confirm whether fluoride was incorporated into the lesions during the remineralization period. From this experiment it can be concluded: (1) clinical photography as currently practiced is not an adequate method of monitoring the remineralization of white spots with large lesion depths and (2) the experimental solution that contains 50 ppm fluoride had a higher efficacy for remineralization than the control solution or the commercial mouth rinse, which contained 225 ppm fluoride.

Crystallography, X-Ray↗

Alveolar ridge augmentation with collagen tubes containing bone and hydroxylapatite.

The use of collagen tube containers with implants of an autogenous bone and particulate hydroxylapatite composite is reported. The tubes were used to shape the mixture and to facilitate its implantation in cases of severe mandibular resorption. Following a minimum of 60 months after implant placement and a minimum of 48 months of denture wear, the implant and overall prosthodontic success were rated favorably by the patient and the prostodontist. Individual implants were evaluated with polarized and fluorescent light microscopy and with microradiography. While the clinical results were good, the microscopic evaluation showed the amount of bone present within the implant to be variable.

Aged↗

A comparative experimental investigation in monkeys between three different implant materials.

To study the tissue integration of 2 new potential biomaterials (whisker silicon carbide, reinforced aluminum oxide, and zirconium oxide strengthened aluminium oxide) a comparative investigation between these ceramic materials and unalloyed titanium was performed on monkeys. Cylindrical implants (4 x 6 mm) were inserted in the femur of 10 adult monkeys. After observation periods ranging from 1 to 8 months the animals were killed and clinical and histological evaluations (microradiography and light-microscopy) were carried out. After 6-8 months, the implants were covered with bone and firmly attached, indicating an uneventful healing. Histologically, a close connection between bone and implant was observed. At some areas the implants were totally osseointegrated and at some areas there were soft tissue layers between implant and bone. No differences in bone repair with regard to the different materials could be observed.

Aluminum↗

Vascular changes after experimentally-induced inflammation in the mandible.

In this study, the vascular supply of an experimentally-induced inflammatory reaction in 5 monkey mandibles is examined. A barium sulfate injection method, combined with contact microradiography was used. In all monkeys, marked vascular changes were seen within the areas of inflammation. Vertical nutrient branches were interrupted shortly after leaving the inferior alveolar artery, and a conglomerate of thin irregular vessels were seen interspersed through newly formed bone trabeculae. The vascular supply to the neighbouring teeth seemed to be unaltered; neither was there any evidence of vascular thrombosis or vasculitis.

Animals↗

Microvascular reanastomozed allogenous iliac crest transplants for the reconstruction of bony defects of the mandible in miniature pigs.

The effects of immunosuppression with cyclosporin A and prednisolone regimens for allogenous iliac crest bone grafts used for mandibular reconstruction were investigated in 40 miniature pigs, for periods of 2, 4 and 16 weeks. Autogenous and allogenous bone grafts without immunosuppression served as controls. Specimens were evaluated by routine histology, direct magnified radiography and fluorescence microradiography. Four out of five autogenous transplants showed a preserved vascular architecture and bony union. None of the allogenous transplants without immunosuppression survived. Primary bone healing of the allografts was noted after short-term immunosuppression. However, occlusion of the nutrient vessels was noted ten days postoperatively. The allografts were not rejected after cessation of the immunosuppressive therapy within an observation period of 4-12 weeks. Revascularization of all areas of the allografts and creeping substitution of the transplanted bone were noted after seven weeks. Infection of the allografts, with failure of bony union, was noted in nine animals, but primary healing of allografts with short-term immunosuppression was demonstrated.

Anastomosis, Surgical↗

Tissue reaction around miniplates used for the fixation of vascularized iliac crest bone grafts.

MATERIAL/METHOD: In six miniature pigs revascularized autologous iliac crest bone grafts were fixed with two 2.0 mm titanium miniplates to a defined defect in the angle of the mandible. Tissue reaction around the implants was examined histologically and histomorphometrically at 2, 4, and 8 weeks after surgery. RESULTS: According to the evaluation of fluorescence microradiography, the cortical and medullary circulation and healing processes of bone in the vicinity of the implanted material were not disturbed after 2 weeks. At 4 weeks after surgery osseous healing occurred at the mandibula/graft interface. The histomorphometric results for the calculated percentage of direct bone/screw contact at 2 weeks were 24% with the grafted bone and 53% with the mandible. Intensive bone formation was observed in all parts of the graft by 4 weeks. The results of direct bone/screw. contact at 4 weeks were 56% and 64% and at 8 weeks 75% and 77% for the grafted and the mandibular bone, respectively. CONCLUSION: With 2.0 mm titanium miniplates used in the animal model, stable osteosynthesis and osseous healing was evident after 8 weeks of surgery. In addition, intensive remodeling of the transplanted bone without signs of creeping substitution was noted, and the bone/screw contact of the miniplate system increased with time.

Animals↗

Characterization of a developing lumbar arthrodesis in a sheep model with quantitative instability.

BACKGROUND CONTEXT: Mechanical forces have been considered responsible for stress shielding an arthrodesis, but the biology of a developing lumbar fusion has not been well characterized. PURPOSE: A large animal model was used to test the hypothesis that mechanical forces modify the biological processes involved in a developing bony fusion. STUDY DESIGN: Lumbar fusion was performed in an ovine model using custom instrumentation that permitted a controlled degree of anterior-posterior translation after surgery. Fusion sites were evaluated by radiography, microradiography, histology and histomorphometry at time points that corresponded with predicted early and later stages of bone healing. METHODS: Fourteen skeletally mature ewes underwent lumbar spinal fusion under general anesthesia. In the control (stable) group, the spine was rigidly fixed with a cage anteriorly and pedicle screws posteriorly. In the experimental (unstable) group, the spine was destabilized by an annulectomy (with no anterior implant) and custom pedicle screws that allowed 2 mm of anterior-posterior translation. Animals were euthanized 6 and 12 weeks after surgery. RESULTS: Radiographs confirmed that the fusion mass had not fully consolidated at either time point. Microradiographs revealed a trend toward increased bone formation at 6 weeks in the stable case as compared with the unstable, but by 12 weeks, this trend had reversed (p=.03). Intramembranous bone formation was the primary mechanism of healing near the transverse process in animals with both stable and unstable fixation. In the area between the two transverse processes, new bone formation occurred primarily through endochondral ossification. At 12 weeks, the stable case had significantly more cartilage formed (p=.023) but less newly formed bone (p=.07) as compared with the quantitatively unstable. CONCLUSIONS: This clinically realistic animal model allowed characterization of the biology of the developing arthrodesis before fusion. Under stable or unstable conditions, endochondral ossification was the predominant mechanism of new bone formation within the intertransverse process region. This finding, which contrasts with previous reports from small animal models of spine fusion, may reflect a difference in biology that results from the increased size of the intertransverse space in sheep as compared with small animals. Interestingly, mechanical instability increased the formation of new bone within this region, but not at the transverse process. Endochondral ossification therefore appears to respond to mechanical factors in the fusion site. The ovine model shows promise as an alternative to the rabbit model and may provide a more stringent test for potential new surgical and nonsurgical strategies for spine fusion.

Animals↗

Alendronate increases bone strength by increasing the mean degree of mineralization of bone tissue in osteoporotic women.

The mean degree of mineralization of bone (MDMB) was measured by quantitative microradiography on transiliac bone biopsies taken from 53 postmenopausal osteoporotic women who had been treated with alendronate (ALN; 10 mg/day) during 2 (9 patients) or 3 years (16 patients) or with placebo (PLA; 15 and 13 patients, respectively). In the same patients, bone mineral density (BMD) values were obtained by dual-energy X-ray absorptiometry of the lumbar spine and femoral neck at the beginning and end of treatment. Histomorphometric parameters and activation frequency of new remodeling units were also measured on the iliac biopsies. After 2 years of ALN, MDMB in compact bone was 9.3% (p = 0.0035) and in cancellous bone was 7.3% (p = 0.0009) higher, respectively, than PLA. After 3 years of ALN, MDMB in compact bone was 11.6% (p = 0.0002) and in cancellous bone was 11.4% (p = 0.0001) higher, respectively, than PLA. After 2 and 3 years of ALN, and compared with the corresponding PLA, the distribution of the degree of mineralization in compact and cancellous bone showed a clear shift toward the highest mineralization values and a decrease in the number of bone structure units having low values of mineralization. The between-group differences in MDMB were similar to those of BMD at the lumbar spine BMD (+8.7% after 2 years and +9.6% after 3 years, respectively), suggesting that MDMB augmentation probably accounted for the majority of the increase in BMD seen with ALN. The data support the hypothesis that the reduction in activation frequency caused by the antiresorptive effect of ALN is followed by a prolonged secondary mineralization that increases the percentage of bone structure units having reached a maximum degree of secondary mineralization and, through this mechanism, MDMB. That these effects contribute to improved bone strength is demonstrated by the reduction in fracture incidence previously demonstrated in these patients.

Absorptiometry, Photon↗

Relationships between trabecular bone remodeling and bone vascularization: a quantitative study.

Beside its well-known role in bone development, vascularization plays a major role in bone cell migration for bone remodeling and metastatic tumor invasion. However, the various techniques used to identify vessels in bone have never been tested for trabecular bone vessel quantification, whereas bone remodeling quantitative parameters are commonly assessed. In this context, we developed and compared various histological techniques used to visualize blood vessels in rat bone in order to quantify them. First, several products were tested by intracardiac infusion to opacify the bone vascular network. The best results were obtained using either an India ink-1% agarose solution or an India ink-saturated barium sulfate solution followed by X-ray microradiography. Second, to identify the types of vessels, we also performed histoenzymology and immunohistochemistry stainings. Neither alkaline phosphatase (for endothelial cells) nor adenosine triphosphatase (ATPase) stainings (for smooth muscle cells) provided a low enough background to allow for vessel identification and quantification. For immunohistochemistry, various specific vessel constituents were analyzed: laminin, smooth muscle cell alpha-actin, factor VIII, and lectin Griffonia simplifolia. Anti-laminin and anti-smooth muscle cell alpha-actin antibodies gave the best results for quantification. Third, after optimization of these techniques, we performed quantitative bone and vessel histomorphometry on two groups of 12 rats each, for which bone remodeling and vessel number and area parameters were measured. No statistical differences were observed between the two groups, confirming the reproducibility of our measurements. A significant relationship was found between vessel number and histodynamic parameters; that is, bone formation rate correlated positively with India ink-positive vessel area (p < 0.009, r2 = 0.54) and alpha-actin-positive vessel number (p < 0.05, r2 = 0.66). Furthermore, we report reproducible techniques for visualization and quantification of vessels in bone that also allowed for simultaneous conventional bone histomorphometry. This methodology should help researchers to better understand the functional and anatomical relationship between trabecular bone and its vascularization during normal or pathological processes.

Adenosine Triphosphatases↗

Adaptive adjustment of the adolescent porcine mandibular condyle.

The effect of occlusal support during primary dentition on the mandibular condyle remains controversial. We sought to determine whether unilateral loss of occlusal support leads to quantifiable adaptive changes of the condyle. Quantitative analysis of condylar growth and spongy bone volume after unilateral removal of teeth on the left side in adolescent minipigs was examined over a period of 4 months. Serial sagittal sections of the temporomandibular joint were examined using microradiography, fluorescence microscopy, and light microscopy. The condyles on the nonextracted side showed a higher growth rate than those on the extracted side, with a 1.56-fold thicker (p = 0.003) additional vertical bone layer. This factor was greater ventrally than dorsally (p = 0.0311), increasing from dorsomedial (1.33) to ventrolateral (2.38). There was therefore a reciprocal change of the condylar surface curve between the left and right condyles. Increased condylar growth correlated with a lower subchondral spongy bone volume (7.38% difference, p = 0.002). The amount of mineralized bone matrix generated was estimated to be about 1.33-fold higher in the nonextraction side condyles compared with those on the extraction side. Thus, unilateral loss of occlusal support was shown to lead to quantifiable alterations of condylar vertical growth and spongy bone volume in minipigs.

Adaptation, Physiological↗