Microradiographic aspect on iliac bone tissue in postmenopausal women with and without vertebral crush fractures.

Bone mass, microstructure and microconnectivity of cancellous bone tissue, microporosity, and microdensity of cortical bone have been evaluated with an image analysing computer on microradiographs in 34 iliac crest bone biopsies from two individual, paired age-matched groups of postmenopausal women. Group 1 comprised 17 cases without vertebral crush fractures (mean age 66 years, range 45-80 years) and time since menopause 20.5 +/- 11.5 years. The second group were 17 cases with vertebral crush fractures (mean age 67 years, range 48-86 years) and time since menopause 16.5 +/- 7.6 years. No differences were observed either in bone mass or in Parfitt's indices of the microarchitecture of their cancellous bone tissue. However, both groups 1 and 2 were different by the number of the structure elements per 30 mm2 of tissue volume and the trabecular bone pattern factor (TBPF) being higher in group 1 than in group 2 (p < 0.005)--showing a better connectivity of bone network in group 2 with crush vertebral fractures than in group 1. The number of terminus to terminus struts of the trabecular bone pattern is increased in group 1 (p < 0.001). On 7-microns stained sections the active cancellous eroded perimeter (Cn.Oc.Pm/B.Pm) is greater in group 1 than in group 2 (p < 0.02). Therefore, in these two populations with a similar degree of osteopenia, a decrease of trabecular connectivity does not provide an explanation of bone fragility and the occurrence of vertebral crushed fractures.

Osteomesopyknosis: report of a new case with bone histology.

A new case of osteomesopyknosis, a rare autosomal dominant axial osteosclerosis is reported, with 4 affected members of the same family. Biochemical investigations, bone mineral content (BMC) measurement, 99mTc HMDP bone scan and microscopy of iliac crest bone and femoral head have been performed on 1 subject. A marked increase of BMC was found, without abnormality of biochemical data. Microscopy of bone showed an increase of trabecular thickness, and a low rate of bone turnover. No abnormality of mineralization was found on microradiographs.

Volume effect on biological properties of a calcium phosphate hydraulic cement: experimental study in sheep.

Injectable calcium phosphate hydraulic cements (CPHC) are a new family of bone substitutes within the class of bone reconstruction biomaterials. In this work, CPHC were tested in two consistencies (preset blocks or liquid paste) in an experimental model of cancellous bone defect in sheep. The defects were eight times larger than those investigated previously in rabbits. Three delays (12, 24, and 52 weeks) were used. Before death, a double label of oxytetracycline and alizarine was made intravenously. The distribution of implants was randomized, histomorphometric evaluation was performed and compared with micrographic observation, and optical microscopy of stained sections was performed either under visible, ultraviolet, or polarized light. The results were compared with spontaneous healing of empty defects and with a control group of normal cancellous bone from sheeps of the same age. No significant difference has been observed between premolded and injected implants. In the sheep model, the degradation and new bone formation rates are three times slower, compared with those observed previously in rabbits. New bone formation increased from 5.9% (12 weeks) up to 11.0% (24 weeks) in the empty defect group. In the cement groups, 28.3% new bone was obtained at 12 weeks, which seemed then to level off (27.8% new bone at 24 weeks). Cement residues appear as radio-opaque cylinders on microradiographs. In all cases, a radiolucent layer was observed at the cement/bone interface at 24 weeks. Stained sections showed the formation of a fibroconnective capsule around the residual cement, which presumably slows down new bone formation. Nevertheless, quantitative bone remodeling was accelerated in the cement group; mineral apposition as well as adjusted apposition rates were higher, and the formation period as well as the mineralization of osteoid tissue were faster compared with empty cavities and controls. These results point to higher osteoblast activity and better exchange with surrounding tissues in the defects filled with cement.

Association of porous hydroxyapatite and bone marrow cells for bone regeneration.

The preparation of hybrid material with osteoinductive capacity may be achieved by association of cultured autologous bone cells with a porous ceramic vehicle. We optimized culture conditions for rabbit marrow stromal stem cells (MSCs), notably by selection from batches of fetal calf serum. Rabbit MSCs formed colony-forming unit-ribroblastic (CFU-Fs) in vitro. Their alkaline phosphatase (ALP) activity was doubled in the presence of dexamethasone. Autologous rabbit serum allowed the formation of ALP-positive CFU-Fs, but results were highly variable depending on the rabbit. We tested the osteogenic potential of autologous cultured (with or without dexamethasone addition in the culture medium) and noncultured rabbit MSCs associated with a porous hydroxyapatite ceramic after a dorsal intramuscular implantation. Nucleated cells (10(7) or 10(8)/mL) were used for the preparation of autologous hybrid material. A significantly higher number of implants containing bone was obtained with a suspension of 10(7) cells/mL cultured in the presence of 10(-8) M dexamethasone. Some positive implants were also obtained with a suspension of 10(8) noncultured cells/mL. We demonstrated the feasibility of preparing rabbit autologous hybrid materials following a process for controlling culture conditions, cell characterization and cell/material association.

Raman microspectrometry studies of brushite cement: in vivo evolution in a sheep model.

Calcium phosphate hydraulic cements are promising synthetic bone grafting materials. Brushite-based cements were implanted for 6 and 12 months in the distal condyle of sheep femur, and their in vivo evolution was investigated by Raman microspectrometry. This new technique can probe small volumes in the cubic micrometer range. Its resolution allows a very fine analysis of crystalline changes in calcium phosphate mixtures at the microscopic level. First, Raman spectra of pure brushite, monetite, and beta-tricalcium phosphate (beta-TCP) were recorded, in order to set a data base for the basic components of brushite cements. These spectra show significant differences in the vibration mode v1 for the phosphate ion (988 and 878 cm(-1) for brushite, 988 and 900 cm(-1) for monetite, 968 and 948 cm(-1) for beta-TCP). These differences are strong enough as to allow the qualitative and quantitative analysis of these crystalline phases in the cement. Implanted sheep femur samples were harvested after 24 and 52 weeks post-op, and prepared for Raman analysis in the form of 1-mm-thick sections. Implants at 24 weeks show a core of residual cement isolated from the surrounding bone by fibroconnective tissue. No trace of brushite was detected by micro-Raman analysis in this area, but instead, a mixture of beta-TCP and Type-B carbonated apatite, the latter being very close in composition and structure to the mineral fraction of normal bone in the vicinity of the implant. Implants recovered after 52 weeks show a decrease of the bone/residual cement perimeter, whereas new trabeculations are formed in the implanted zone; the small amounts of residual cement still present are substantially transformed into Type-B carbonated apatite containing small amounts of proteins. In the same area, some beta-TCP particles are also detected showing that, contrary to brushite, the excess beta-TCP originally present in the cement is not completely metabolized. In the implanted zone already converted into trabecular bone, Raman microspectrometry shows the characteristic spectrum of normal bone.

Fate of bioresorbable poly(lactic acid) microbeads implanted in artificial bone defects for cortical bone augmentation in dog mandible.

The fate was examined of poly(lactic acid) microbeads implanted in large artificial defects created in cortical bone of dog mandibles. Two poly(lactic acid) polymers--poly(L-lactic acid) (PLA 100) and poly(DL-lactic acid) (PLA 50)--were used to make microbeads by solvent evaporation with poly(vinyl alcohol) as surfactant. Histological observation of non-decalcified mandibular bone showed that no real bone regeneration existed in the experimental bone defects 18 months after PLA 100 microbeads implantation. The same observation was made 6 months after implantation of PLA 50 microbeads. PLA 100 and PLA 50 microbeads appeared unable to induce regeneration of cortical bone defects of dog mandible, in contrast to previous observations in man for PLA 50 large implants. The failure is tentatively assigned to the presence of poly(vinyl alcohol) at the surface of microbeads.

Influence of polymeric additives on the biological properties of brushite cements: an experimental study in rabbit.

The resorbability and ability of calcium phosphate hydraulic cements to promote new bone formation was investigated in vivo. The effects of two hydrosoluble polymeric additives (hyaluronic acid, and xanthan gum,) on the biological response of two brushite cement formulations (BHC-A vs BHC-B) was investigated. The brushite cements differed in P/Ca (0.71 vs 0.98) and S/Ca (0.10 vs 0.005) atomic ratios and by the presence of calcium sulfate hemihydrate in BHC-A. Polymer-free cements were used as controls. Cement specimens were injected in cylindrical bone defects manually drilled in the distal condyle of rabbit femora. The implants were harvested at 12 and 24 weeks after implantation and subjected to quantitative histomorphometry. The study showed a significantly lower resorption rate for cement BHC-A, which induces the formation of well-mineralized bone in close apposition to the residual material. In contrast, cement BHC-B showed a significant increase of bone formation period and the formation of a thick layer of unmineralized osteoid tissue at the bone/residual cement interface. The presence of xanthan gum made the biological response even worse, particularly in the case of cement BHC-B. The presence of hyaluronic acid has little effect, except for a slight decrease in initial resorption rate, in the case of cement BHC-A.

Experimental model of posterolateral spinal arthrodesis in sheep. Part 1. Experimental procedures and results with autologous bone graft.

OBJECTIVES: The authors evaluated the reliability in obtaining a posterolateral spinal arthrodesis (PSA) with autologous bone graft. SUMMARY OF BACKGROUND DATA: Posterolateral spinal arthrodesis using autogenous cancellous bone graft is the most simple and efficient technique to get a spinal graft. No extensive biomechanical study of PSA is available. Thus, an experimental model of PSA is needed. METHODS: Eleven sheep underwent lumbar autologous bone grafts and Cotrel-Dubousset instrumentations, and four sheep were used as controls. Sacrifice and biomechanical evaluation of the lumbar spines were performed after 1 year. RESULTS: All grafts appeared continuous. A large decrease of flexibility (in rotation and in translation) was found in grafted spines in every direction. Failure in extension occurred at a mean value of 35.26 +/- 3.71 Nm. CONCLUSION: A constant and homogenous PSA appears to be obtained in sheep under conditions close to the human surgery.

Experimental model of posterolateral spinal arthrodesis in sheep. Part 2. Application of the model: evaluation of vertebral fusion obtained with coral (Porites) or with a biphasic ceramic (Triosite).

OBJECTIVES: The authors evaluated two bone substitutes in a posterolateral spinal arthrodesis (PSA) model in sheep: coral porites (99% calcium carbonate, Biocoral, Inoteb, France) and a biphasic ceramic (BCP) (65% hydroxyapatite and 35% B tricalcium phosphate, Triosite, Zimmer International). SUMMARY OF BACKGROUND DATA: Bone substitutes would be of great interest for PSA. Previous trials began with two kinds of biomaterials: natural coralline calcium carbonate, and phosphate calcium ceramic. METHODS: A lumbar PSA was performed in 11 sheep (coral group) and in 9 sheep (BCP group). Sacrifice and biomechanical tests were performed after 1 year. RESULTS: A large decrease of flexibility in all directions was obtained with both coral PSA and BCP PSA similarly to autologous graft. No nonfusion case was observed. CONCLUSION: In conditions close to the human surgery, a PSA can be obtained using either coral porites or BCP as bone substitutes.

Migration of polyethylene debris along well-fixed cemented implants.

Implants, consisting of smooth Inox cylinders, were cemented into the lower femur and upper tibia of nine sheep to study the distal migration of polyethylene particles. Some implants had a titanium-bead porous coat at the proximal end. These were of three types: In the first type, the porous coat was covered with hydroxyapatite to obtain a bony seal; the second type was prepared for a polymethylmethacrylate seal; in the third type, the porous zone was surrounded by a 2-mm-thick space to allow the formation of a fibrous seal. Small polyethylene particles were injected into the knees once a week during the third and fourth months after implantation. The animals were euthanized 2 months later. Major longitudinal sections of the implants and the surrounding bone were examined under a polarized light microscope. Birefringent particles were counted at the cement-bone and cement-implant interfaces. Osteolysis was not observed. None of the seals significantly decreased the migration of particles around the cemented part of the implants. Particles were observed in cement fissures and vacuoles. They migrated at both interfaces and in the bone itself. They were visible in marrow spaces between bone trabeculae.

Tissue engineering and skeletal diseases.

Tissue engineering, a cross between the science of the living organism and that of engineering, aims to replace, maintain or improve human tissue functions, by means of tissue substitutes containing living elements. Thus, it is about production of artificial tissue, using (alone or in combination) cells, matrix or bioactive factors. Their association gives rise to a hybrid biomaterial combining biological components (cells, growth factors or adhesion proteins) and materials (polymers, ceramics). The applications are wide-ranging, from the skin, to the liver, or to the cornea as well as to the locomotor system. Bone tissue engineering has advanced the most in this field, partly because of the progress made by research into bone substitutes, although cartilage and tendons are also concerned. This technology requires cell culture (committed cells or more often bone marrow stem cells), biomaterials (porous materials with controlled architecture and cements), growth factors (such as 'Bone Morphogenetic Proteins'), the proteins implicated in cell adhesion (such as fibronectin or the aminoacid sequences specifically recognised by integrin subunits) or gene therapy (notably using transfected stem cells). Tissue engineering and regenerative stimulation of tissue are now booming on experimental and industrial levels and clinical applications are increasingly numerous. Considering the potential of these technologies, they should continue to develop widely.

[Structural MRI study of the Achilles tendon. Correlation with microanatomy and histology]. / Etude structurale du tendon d'achille en IRM. Corrélation avec la microanatomie et l'histologie.

On T1 and T2 weighted MR Imaging, normal Achilles tendon is generally described as a homogeneous low signal structure. However, punctuate and/or short linear high signal foci are often seen, especially on the anterior part of axial sections. These images are not artifacts. The aim of this study was to correlate MR images of 20 normal Achilles tendons with micro-anatomic and histologic studies of 2 cadaver tendons. Intratendinous vessels were found in connective tissue septa called mesotenon. Foci of high signals result from this mesotenon. On T1 weighted images, a normal Achilles tendon can present high signal images without any disease process.

[Microradiography in the study of trabecular parameters]. / La microradiographie dans l'étude des paramètres trabéculaires.

The intermethod variation in measurement of trabecular bone volume (VTO) and the indirect estimation of its microstructure according to Parfitt MTPT (microns), MTPD (/mm) and MTPS (microns) were evaluated in seven undecalcified bone biopsies by analyzing the microradiograph of a 100-microns-thick section with an automatic method (IBAS II Zeiss, Munich) in addition to reference methods (manual and semiautomatic) described in the literature and performed on 7-microns-thick stained sections. Three consecutive 7-microns-stained sections and one 100-mu-thick microradiographed section were taken in each specimen. The 100 microns-thick section was also superficially stained. Trabecular bone volume was measured with both a manual integrating eyepiece and an automatic method. The automatic method on the microradiograph underestimated the VTO by 24.42%. There was a correlation (r = 0.75; p < 0.02) between the manual and computerized methods. Mean trabecular plate thickness (microns), mean trabecular plate density (/mm) and mean trabecular separation (microns) were measured with both semiautomatic and automatic methods. The automatic method on the microradiograph underestimated the MTPT (microns) by 18.98% and the MTPD (/mm) by 14.14% and overestimated the MTPS (microns) by 23.17%. For the MTPT (microns) there was a correlation (r = 0.88; p < 0.02), between both methods, and the correlation was good for MTPD (/mm) (r = 0.97; p < 0.001), and MTPS (r = 0.86; p < 0.002).

[Fatigue rupture of a Harris-Galante shell in contact with an autograft of the femur head. Analysis of bone, mechanical and metallurgical factors]. / Rupture en fatigue d'une cupule de Harris-Galante au contact d'une autogreffe de tête fémorale. Analyse des facteurs osseux, mécaniques, et métallurgiques.

We observed, after 38 months of follow-up, the rupture of the metallic shell of a Harris-Galante cup implanted in a dysplastic acetabulum augmented with a femoral head autograft. Bone ingrowth was extended to 53% of the porous surface, but only in the areas in contact with the true acetabulum. No bone ingrowth was identified in the fiber mesh in contact with the autograft. The graft was necrotic on histologic examination and showed collapse radiographically. These last conditions were responsible for shear stress in the part of the cup that was in contact with the graft-acetabulum junction. These stresses were involved in the fatigue rupture mechanism identified on microscopic examination of rupture surfaces. We observed metallic structure anomalies in the failed cup by comparing with another Harris-Galante cup considered as a reference: larger alpha elements, reduction of the titanium equiaxial structure. These defects could be related to uncontrolled temperature during the sintering process utilized for fiber mesh fixation. These structural anomalies, by reducing the metallic fatigue strength, potentiated the deleterious effect of partial bone ingrowth and graft collapse.

[Femoral lengthening by transverse diaphyseal osteotomy. Experimental study in the dog]. / Allongement fémoral par ostéotomie diaphysaire transversale. Etude expérimentale chez le chien.

Previous work has shown that extensive areas of bone necrosis developed after Z-osteotomies of the femoral shaft. A study was made on 11 dogs to compare the effects of a transverse osteotomy followed by immediate lengthening, stabilisation by plates and bone grafting. The bone vitality was studied at 8 and 16 weeks by histological sections and angiography. Attention was focussed on the area of lengthening and the bone ends. The results showed the superiority of transverse osteotomy over Z-osteotomy. Bone necrosis diminished at the 16 th week. In the area of lengthening, the vascularity of the grafts was satisfactory. In contrast, the tendons of the Z-osteotomy showed constant necrosis.

[Patellar reconstruction during total knee arthroplasty after previous patellectomy]. / Reconstruction patellaire au cours d'une arthroplastie totale sur genou patellectomisé

PURPOSE OF THE STUDY: In order to limit the consequences of prior total patellectomy on knee function after total knee arthroplasty, we propose an original technique using an en bloc patellar graft to reconstruct the quadriceps lever arm. The purpose of this work was to describe the technique and present preliminary results obtained in seven patients. PATIENTS AND METHODS: Seven en bloc patellar grafts were performed in seven patients (6 women and 1 man) during total knee arthroplasty procedures for femorotibial degenerative joint disease after prior patellectectomy (mean 13 years before arthroplasty, range 5-20 years). We implanted three posterior stabilized prostheses, one ultracongruent prosthesis, two prostheses preserving the posterior cruciate ligament, and one bi-cruciate prosthesis. The patella was reconstructed with an autologous bone graft fashioned from the tibial plateau in six cases, and with a homologous graft fashioned from a bone-bank femoral head in one. Transosseous sutures were used to fix the graft to the extensor system. A polyethylene button was cemented on five of the grafts. RESULTS: Active extension was defective in five knees prior to the patellar graft and in only one knee after patellar reconstruction. The IKS knee score improved from 41 points (range 35-45) before surgery to 78 points (range 55-95) after grafting and the IKS function score from 35 (range 5-50) to 72 (range 40-100). Four of the seven grafts were removed (eight weeks to five years after implantation, mean two years) mainly due to failure of fixation leading to progressive migration. Follow-up varied from eight weeks to six years. Mean follow-up for the three grafts still in place was 4.6 years (4-6 years). After removing the patellar graft (4 knees) the IKS knee score decreased to 68 points (20-95) and the IKS function score to 62 points (30-100). Lack of active extension reappeared in three of the knees after removing the patellar graft but was not observed in the three knees with the patellar graft in place (and flexion was at least 110 degrees ). Microradiography of the patellar graft explanted after two years revealed peripheral corticalization with areas of living bone tissue seen on the pathology specimens. Inversely, the autografts explanted at eight weeks and at five years exhibited bone necrosis. CONCLUSION: This preliminary study shows that patellar autograft can improve the performance of total knee arthroplasty on patellectomized knees. The fixation technique must be improved using transosseous transverse sutures in addition to peripheral sutures in order to limit secondary migration of the graft. A patellar autograft fixed into the extensor system can remain viable two years after implantation. This technique can be proposed when total knee arthroplasty is indicated for a patellectomized knee, particularly in patients with lack of active extension.

[Development of a model which makes it possible to test injectable bone substitutes and evaluation of a calcium phosphate cement]. / Développement d'un modèle permettant de tester des substituts osseux injectables et mise au point d'un ciment phosphocalcique de comblement.

New percutaneous filling techniques are beginning to be used in bone tumor pathology. The purpose of this study was to develop a reproducible model for testing new injectable bone substitutes. A closed cancellous bone defect was created in the distal femoral condyles of rabbit. Bone defect was filled by a percutaneous injection. Several situations were tested: spontaneous evolution of unfilled cavities, evolution of cavities filled either with a conventional orthopedic cement (PMMA), either with a mixed collagen-hydroxyapatite material, either with a biomaterial derived from the vegetal protein zein, either with a calcium phosphate cement brushite type. The new bone formation was quantified in the defect for each group in which several delays were tested. A bone defect of reproducible size was obtained consistently. A partial bone formation was observed in the unfilled group. The best bone formation rate was obtained in the phosphate cement group. This model showed the interest of injectable biomaterials and more precisely the interest of the phosphate calcium cements as brushite.

[Peri-osteocytic calcium fixation under calcitonin treatment. Microradiographic study using automatic analysis of images in 8 cases of osteogenesis imperfecta]. / Fixation calcique péri-ostéocytaire sous calcitonine. Etude microradiographique à l'analyseur automatique d'images dans huit cas d'ostéogénèse imparfaite

The authors treated eight cases of osteogenesis imperfecta using doses of 1 to 2 units of MRC over periods ranging from 6 weeks to 27 months. The lacunar surface area measured before and after treatment was greatly decreased as a result of the latter, probably as the result of marked calcium fixation around the osteocytes, though this effect seems to be transient. The hypothesis of an inactivation of calcitonin on a immunological basis or of a secondary hyperparathyroid reaction is suggested.

[Microradiographic study of iliac bone biopsies taken after treatment of postmenopausal osteoporosis with sodium fluoride. Histomorphometric correlations]. / Etude microradiographique de biopsies osseuses iliaques prélevées après traitement de l'ostéoporose post-ménopausique par le fluorure de sodium. Corrélations histomorphométriques.

Thirty seven female patients with osteoporosis underwent iliac bone biopsy after 10 to 23 months of continuous or discontinuous treatment with sodium fluoride, calcium, and vitamin D. Microradiographs of the biopsies from 13 patients showed one or--more often--several of the characteristic appearances of bone fluorosis: defects in periosteocytic mineralization, unmineralized strips inside the trabeculae of spongy bone, construction of fibrous bone, hypercalcified periosteal apposition. The occurrence of these microradiographic signs of fluorosis does not seem to be linked either to the age of the patients, to the interval elapsed since ovarian function ceased, or to the degree of bone remodelling at the time that treatment was started. Moreover, none of these patients showed signs of even moderate renal insufficiency. Should the development of this fluorosis, which is most frequently clinically asymptomatic, be considered as a condition or as a factor in therapeutic efficacy? Or are the effects detrimental and therefore to be avoided by a reduction in dosage or a change in therapeutic methods?