The organization of the inorganic crystals and organic matrix in bone and dental enamel.
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Bone and Bones↗
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[Bone transplantation in diaphyseal defects of the long bones].
The treatment of large diaphyseal defects has remained in the domain of autologous bone transplantation, but has been substantially complemented by the Ilazarow technique. Bone formation following transplantation or segmental transport is controlled by three interacting mechanisms involving osteoregeneration, osteovascularization and osteostabilization. All three components are equally important in the healing process of large cortical defects. Osteoregeneration is inseparably linked to an intact bone matrix. Therefore, entirely inorganic bone substitutes are not suitable for difficult conditions such as is the case diaphyseal defects.
Adolescent↗
Osteoporosis and periodontal disease: is there a relationship?
Osteoporosis, by definition, is a generalized progressive reduction in both bone mineral and bone matrix which results in bone of normal composition but decreased mass. Functionally, osteoporotic bone is characterized by greater fragility and an increased propensity to fracture. It ranks as the most common metabolic bone disease and the most common skeletal disorder in the world. As such, it constitutes a major public health problem. Due to the extent of the disease, many have questioned its relevance to the maxilla and mandible and its possible relationship to periodontitis. The purpose of this paper is to review both osteoporosis and periodontitis and to present the research completed to date which has investigated the possible interrelationships between the two diseases.
Age Factors↗
[Normal bone remodelling--what can go wrong in osteoporosis?].
The skeleton is a metabolically active organ in constant change and renewal. Specialised cells are responsible for resorbing and synthetizing bone matrix. The process of bone remodelling, by removing old and damaged bone tissue and replacing it with new, is necessary for conserving the integrity of bone. In recent years, research has given new insight into the fine relationship between different cell types in bone tissue. This has enhanced our understanding of the pathophysiology behind osteoporosis and other metabolical bone diseases. This article gives a summary of our present knowledge of the normal cell biology of bone and what contributes to its regulation. How this helps the understanding of normal and pathological bone loss is illustrated. It is important to understand bone remodelling in order to interpret measurements of bone mineral density and biochemical markers accurately.
Aged↗
Collagen and non-collagenous proteins in different mineralization stages of human femur.
Mineralized tissues exhibit varying degrees of mineralization in different areas within the same bone. Using the technique of density gradient fractionation, bone powder from the diaphysis of human femur has been separated in different fractions corresponding to the degree of mineralization. Isolated bone fractions were analysed for their content in collagen and non-collagenous proteins. The results showed marked differences between compact and spongy bones, this latter containing higher proportions of little mineralized bone particles than the former (p less than 0.01). The ethylenediaminetetra-acetic acid (EDTA) extractability and the bone matrix size decreased relative to the decrease in specific gravity of bone particles. Among the matrix components of different fractions, sialoprotein consistently increased with the increase in specific gravity while proteoglycan decreased in reverse manner to the increase in collagen. However, in the most mineralized fraction (specific gravity: 2.33 g/cm3), the proteoglycan amount increased while collagen decreased. In conclusion, this study of bone maturation in human femur confirms the suitability of the technique of density gradient fractionation in the studies of bone matrix-mineral interactions. Apart from the fraction with the highest specific gravity, the analytical results obtained in fractions are similar to those observed in age-related bone changes, suggesting that the increase in mineralization degree of bone particles may be related to their age.
Aging↗
Autogenous cultured bone graft--bone reconstruction using tissue engineering approach.
Maniatopoulos et al. reported the formation of calcified bone-like tissue when rat bone marrow cells were cultured in the presence of dexamethasone and beta-glycerophosphate. We have succeeded to construct the in vitro cultured bone on the porous framework of hydroxyapatite ceramics (HA). After 2 weeks of the culture, the construct showed the existence of mineralized collagen fibers on the surface of HA determined by Scanning Electron Microscopy. The construct also demonstrated high alkaline phosphatase (ALP) activity together with noticeable level of osteocalcin. The results indicate that the construct consisted of thin layer of bone matrix covered hydroxyapatite surface and abundant bone forming active osteoblasts. After the in vivo implantation of the construct, volume of the matrix increased and obvious bone tissue was detected by ordinal microscopy even one-week after implantation and its high osteogenic activity was maintained for a long term (one year). The in vivo bone is biologically active tissue evidenced by Northern blot analysis of the implanted construct which showed ALP and osteocalcin mRNA expression comparable to those of normal cancellous bone. These results demonstrate that the in vitro fabricated cultured bone/HA construct can possess new bone forming capability in in vivo situations. We have also succeeded to fabricate the construct using aged human marrow cells and the construct showed thick lamellar bone formation after the in vivo implantation. Based on these findings, we propose alternative approach for bone reconstruction surgery using the autogenous cultured bone/HA construct. Importantly, we can fabricate the implantable autogenous bone tissue derived from patient's marrow cells and the cells can be obtained by needle aspiration without damaging the patient's normal tissue.
Animals↗
A histological evaluation of the involvement of Bio-Oss in osteoblastic differentiation and matrix synthesis.
This study was designed to investigate the responses of bone cells to a deproteinized bovine bone material, Bio-Oss (Geistlich-Pharma, Wolhunsen, Switzerland), which was grafted in artificial bone defects of rat femurs. Standardized bone defects in the cortical bone of the right femurs were grafted with Bio-Oss particles. Narrow penetrations were prepared on the bottom of the cavity, enabling osteogenic cells to migrate from the bone marrow. A defect in the left femur without Bio-Oss was used as a control. The treated femurs were histochemically examined at 1, 3, 5, 7, and 14 days after the operation. At day 1, no osteogenic migration into the cavities occurred in either the control or experimental groups. At day 3, alkaline phosphatase (ALPase) immunohistochemistry showed a migration of the positive cells at the bottom of the cavities of the experimental groups, but not in the control ones. At day 5, new bone formation was recognized at the bottom of the cavity of both groups. In the experimental group, ALPase-positive cells were localized on Bio-Oss and/or on the thin bone matrix that covered this material. The superficial layer of Bio-Oss underlying the newly formed bone exhibited osteocalcin immunoreactivity. Transmission electron microscopy revealed osteoblasts depositing bone matrices--including collagen fibers--on the surface of Bio-Oss. At days 7 and 14, woven bone occupied the previous cavities of both control and experimental groups, accompanied by osteoclasts. Thus, Bio-Oss appears to serve as a scaffold for osteogenic cells as well as to promote osteoblastic differentiation and matrix synthesis.
Alkaline Phosphatase↗
Non-enzymatic glycation of bone collagen modifies osteoclastic activity and differentiation.
Type I collagen, the major organic component of bone matrix, undergoes a series of post-translational modifications that occur with aging, such as the non-enzymatic glycation. This spontaneous reaction leads to the formation of advanced glycation end products (AGEs), which accumulate in bone tissue and affect its structural and mechanical properties. We have investigated the role of matrix AGEs on bone resorption mediated by mature osteoclasts and the effects of exogenous AGEs on osteoclastogenesis. Using in vitro resorption assays performed on control- and AGE-modified bone and ivory slices, we showed that the resorption process was markedly inhibited when mature osteoclasts were seeded on slices containing matrix pentosidine, a well characterized AGE. More specifically, the total area resorbed per slice, and the area degraded per resorption lacuna created by osteoclasts, were significantly decreased in AGE-containing slices. This inhibition of bone resorption was confirmed by a marked reduction of the release of type I collagen fragments generated by the collagenolytic enzymes secreted by osteoclasts in the culture medium of AGE-modified mineralized matrices. This effect is likely to result from decreased solubility of collagen molecules in the presence of AGEs, as documented by the reduction of pepsin-mediated digestion of AGE-containing collagen. We found that AGE-modified BSA totally inhibited osteoclastogenesis in vitro, most likely by impairing the commitment of osteoclast progenitors into pre-osteoclastic cells. Although the mechanisms remain unknown, AGEs might interfere with osteoclastic differentiation and activity through their interaction with specific cell-surface receptors, because we showed that both osteoclast progenitors and mature osteoclasts expressed different AGEs receptors, including receptor for AGEs (RAGEs). These results suggest that AGEs decreased osteoclast-induced bone resorption, by altering not only the structural integrity of bone matrix proteins but also the osteoclastic differentiation process. We suggest that AGEs may play a role in the alterations of bone remodeling associated with aging and diabetes.
Adult↗
Repair of intra-membranous bone fractures and defects in rats. Immunolocalization of bone and cartilage proteins and proteoglycans.
Osseous healing of experimental fractures and defects in membranous bone was studied in an animal model and the appearance and localization of selected bone and cartilage proteins and proteoglycans determined by polyclonal antibodies. The bone lesions were made in the parietal bone of young rats and subsequently studied on days 3, 5, 8, 15, 30, and 100 after surgery. The bone matrix proteins investigated (62 kDa, bone sialoprotein I and II, and osteopontin) appeared early, adjacent to the periosteal surfaces (pericranium and dura mater) and the marginal bone. The staining reactions were maximal at days 8 or 15 after trauma. Similar patterns were discerned for some cartilage macromolecules studied (58 kDa, 59 kDa, and chondrocalcin), although others showed no labelling whatsoever (148 kDa, and 400 kDa proteins). The proteoglycans PG-S1, PG-S2, and PG-LA were not identified. No callus or cartilage formation were associated with the bone healing process, and the differences between the regenerative pattern of the fractures and defects were limited. The findings emphasize the importance of rigid fixation in craniomaxillofacial surgery.
Animals↗
Dentin matrix protein 1 is expressed in human lung cancer.
UNLABELLED: We have previously shown that breast and prostate cancers express bone matrix proteins. DMP1 expression was evaluated in 59 human lung cancer samples at the protein and mRNA levels. It was detectable in 80% of the cases, suggesting a potential role for DMP1 in tumor progression and bone metastasis. INTRODUCTION: Previously, we and others have shown that bone extracellular matrix proteins such as bone sialoprotein (BSP) and osteopontin (OPN) are expressed in various types of cancer that are characterized by a high affinity for bone including breast, prostate, and lung adenocarcinoma. Based on biochemical and genetic features, BSP, OPN, dentin matrix protein 1 (DMP1), and dentin sialophosphoprotein (DSPP) have been recently classified in a unique family named SIBLING (small integrin-binding ligand, N-linked glycoprotein). Therefore, we investigated whether DMP1 could also be detected in osteotropic cancers. MATERIALS AND METHODS: We first used a cancer array for evaluating the relative abundance of DMP1 transcript in a broad spectrum of human cancer tissues. This screening showed that DMP1 was strongly detectable in lung tumors compared with normal corresponding tissue. In a second step, we used an immunophosphatase technique and a specific polyclonal antibody directed against DMP1 to examine the expression of DMP1 in 59 human non-small cell lung cancer samples, including 29 squamous carcinoma, 20 adenocarcinoma, and 10 bronchioloalveolar carcinoma. Student's t-test was used to determine the statistical significance of immunostaining scores between the lung cancer histological groups studied and between cancer and normal lung tissues. RESULTS: Our results show that DMP1 is detectable in 90% of the adenocarcinoma and squamous carcinoma analyzed while 8 of 10 bronchioloalveolar specimens were negative. DMP1 immunostaining intensity and extent scores were significantly higher in adenocarcinoma (p = 0.0004) and squamous carcinoma (p < 0.0001) samples compared with adjacent normal lung tissue. In situ hybridization experiments confirmed that DMP1 mRNA is localized in lung cancer cells. CONCLUSION: In this study, we show that a third SIBLING protein is ectopically expressed in lung cancer. The role of DMP1 in lung cancer is largely unknown. Further studies are required to determine the implication of this protein, next to its sisters SIBLING proteins, in tumor progression and bone metastasis development.
Adenocarcinoma↗
The effect of protein-energy malnutrition on the development of bones in newborn rats.
Collagen synthesis, 45Ca uptake, and the half-life of 45Ca in mandibles and long bones of rat pups suckled on dams fed 25 or 6% protein diets were determined. Collagen synthesis in both mandibles and long bones was impaired in the malnourished group at an early age. Protein-energy malnutrition appears to affect different developmental processes in these two bones. The principal interference with collagen synthesis in the mandible occurs when bone proline is converted into hydroxyproline. In contrast, proline uptake from the blood for deposit in bone matrix is the step of collagen synthesis mainly affected in long bone. In addition, the critical growth period of mandible was different from that of long bone. The calcium complex fraction (calcium bound to protein plus that in inorganic salts) was the main compartment affected in the malnourished group. However, the efficiency of calcification per milligram of matrix in the calcium complex fraction of the mandible and long bone was approximately the same in the control and malnourished groups, thus suggesting that accumulation of calcium occurs in parallel with the formation of bone matrix regardless of the nutritional conditions. The half-life of 45Ca in the various calcium fractions of both types of bone was 72 hours in both the control and malnourished groups except the calcium complex portion of the long bone of the control group, which was about 100 hours.
Aging↗
Transitions in collagen types during endochondral ossification in human growth cartilage.
Immunohistochemical staining for Types I and II collagen in human growth cartilage showed that cartilage matrix consists of Type II collagen, whereas bone matrix contains Type I collagen. Endochondral ossification began with the deposition of Type I collagen by cells derived from bone marrow on the surface of the eroded cartilage. However, territorial matrix of the last hypertrophic chondrocytes contained both Types I and II collagen, thereby indicating that the degenerating chondrocytes initiate the synthesis of Type I collagen. This matrix, consisting of Types I and II collagen, is then replaced by newly formed osteoid.
Bone Development↗
Lung, heart, and kidney express high levels of mRNA for the vitamin K-dependent matrix Gla protein. Implications for the possible functions of matrix Gla protein and for the tissue distribution of the gamma-carboxylase.
We have used cDNA probes for two small vitamin K-dependent bone matrix proteins, bone Gla protein (BGP) and matrix Gla protein (MGP), to evaluate the possibility that either of these proteins might be synthesized by the various soft tissues previously shown to have gamma-carboxylase activity. BGP mRNA was found in bone but not in any of the soft tissues tested, a result which reinforces the view that plasma BGP is a specific marker for bone metabolism. In contrast, MGP mRNA was found in all rat tissues examined. Lung and heart have 10-fold higher levels of MGP mRNA than bone, and kidney has a 5-fold higher level. Despite the high levels of MGP mRNA in heart and kidney, these tissues contain 40-500-fold lower concentrations of MGP protein than bone. Immunofluorescence was used to identify cells that contain MGP in kidney, lung, heart, and spleen. In each tissue, MGP was found in discrete tissue-specific cell types. In most of the soft tissues tested, MGP is the first well characterized substrate for the vitamin K-dependent carboxylase found to be synthesized. The exceptionally broad tissue distribution for MGP synthesis demonstrates that the function of MGP is not specific to connective tissues, and the low levels of MGP antigen in soft tissues with high MGP mRNA levels indicate that MGP is unlikely to act solely by virtue of its accumulation in an extracellular matrix.
Animals↗
Immunolocalization of matrix metalloproteinase-13 on bone surface under osteoclasts in rat tibia.
Matrix metalloproteinase (MMP)-13 (an interstitial collagenase also called collagenase 3) is involved in degradation of extracellular matrix in various tissues. Using immunohistochemistry and Western blotting, we investigated localization of MMP-13 in rat tibia, to clarify the role of MMP-13 in bone resorption. MMP-13 reactivity was mainly seen on bone surfaces under osteoclasts, and in some osteocytes and their lacunae near osteoclasts. However, immunoreactivity was not seen in chondrocytes or osteoclasts. MMP-13 was also localized on cement lines in the epiphysis. In the growth plate erosion zone, perivascular cells showed MMP-13 reactivity. Immunoelectron microscopy revealed that MMP-13 was localized on the bone surfaces, under the ruffled borders and some clear zones of osteoclasts. Gold-labeled MMP-13 was closely associated with collagen fibrils. Gold labeling was also detected in Golgi apparatus of osteocytes adjacent to osteoclasts and bone lining cells. Western blotting showed that MMP-13 was mainly associated with mineralized bone matrix. These findings suggest that MMP-13 synthesized and secreted by osteoblast-lineage cells is localized under the ruffled borders of osteoclasts. MMP-13 may play an important role in degradation of type I collagen in bone matrix, acting in concert with cathepsin K and MMP-9 produced by osteoclasts. MMP-13 in perivascular cells may be involved in removal of cartilage matrix proteins such as type II collagen and aggrecan.
Animals↗
Effects of enamel matrix derivative on mineralized tissue formation during bone wound healing in rat parietal bone defects.
Enamel matrix derivative (EMD: Emdogain) has been reported to stimulate the biosynthesis and regeneration of trabecular bone. To address whether the biological action of EMD is dependent on the local environment of osseous tissue, circular perforations were made in parietal bones and immediately filled with either EMD or its carrier, propylene glycol alginate (PGA), as control. On post-operative days 4-60, the dissected bones were examined by various histological techniques. New bone matrix, which was immunoreactive for bone sialoprotein (BSP), was formed from the periosteum at the peripheral area of perforations. Different from the findings reported in injured long bones, mineralized tissue was produced in the regenerating connective tissue within bone defects. This mineralized tissue was hardly immunostained for BSP, contained few collagen fibres, and lacked osteocytic lacunae and layers of osteoblasts and osteoid. Energy-dispersive X-ray analysis showed that Ca and P weight % and Ca/P molar ratio of this mineralized tissue were similar to or slightly higher than those in the pre-existing parietal bones. In addition, most multinucleated cells located in mineralized tissue lacked a ruffled border structure and showed weak immunoreaction for the lysosomal cysteine proteinase, cathepsin K, whereas those located in the bone matrix exhibited ruffled borders and strong cathepsin K expression. However, multinucleated cells located in both tissues were strongly stained for tartrate-resistant acid phosphatase. The volume fraction of such mineralized tissue appeared to be higher in EMD-applied bones than in PGA-applied controls. The mineralized tissue-forming stromal cells within bone defects appeared to show greater accumulation in EMD-applied bones than in PGA-applied controls. Our results suggest that the bioactive effects of EMD on bone wound healing and mineralized tissue formation depend, at least in part, on the local osseous environment where EMD has been applied.
Animals↗
Effects of enamel matrix derivative to titanium implantation in rat femurs.
The effects of enamel matrix derivative (EMD; Emdogain) on new trabecular bone induction after pure bioinert titanium (Ti) implantation in the rat femur were examined by means of routine light and transmission electron microscopy, immunohistochemistry, and backscattered electron image analysis. Newly designed mini-Ti implants (3.5 mm in length and 1.6 mm in diameter) were placed in the corticotrabecular area of the femur with either EMD or its carrier, propylene glycol alginate, as control. On post-implantation days 4, 7, 14, and 30, the dissected femur was examined in the transverse direction through Ti implants. In both control and EMD-applied femurs, trabecular bone formation was recognized over the implant surfaces and within medullary cavities even at 4 days post-implantation. These newly formed bone trabeculae around the Ti implants were immunoreactive for bone sialoproteins as a bone matrix marker, and osteoclastic bone resorption became evident in these bone trabeculae after 7 days post-implantation. Although trabecular bone area around the implants was markedly decreased at 30 days post-implantation compared with those at 14 days, the trabecular bone areas in EMD-applied femurs were significantly greater than those in propylene glycol alginate-applied femurs at both 14 and 30 days post-implantation. Our results suggest that EMD is an effective biological matrix for enhancing new trabecular bone induction and resulting attachment of orthopedic prostheses to the recipient bone.
Animals↗
Multi-center clinical comparison of combination anorganic bovine-derived hydroxyapatite matrix (ABM)/cell binding peptide (P-15) and ABM in human periodontal osseous defects. 6-month results.
BACKGROUND: Intraosseous periodontal defects present a particular treatment problem. New bone replacement grafts offer promise for improved results. METHODS: The role of a synthetic cell-binding peptide (P-15), combined with anorganic [corrected] bovine-derived hydroxyapatite bone matrix (ABM), was compared to ABM alone in human periodontal osseous defects in a controlled, monitored, multi-center trial. Following appropriate initial preparation procedures, flap surgery with defect and root debridement was performed. Two osseous defects per patient were treated randomly with each procedure after surgical preparation. Appropriate periodontal maintenance schedules were followed, and at 6 to 7 months, re-entry flap surgery was performed for documentation and finalization of treatment. RESULTS: T test and Mann-Whitney U analyses of patient mean values from 33 patients revealed that the combination ABM/P-15 grafts demonstrated significantly better mean defect fill of 2.9 +/- 1.2 mm (72.9%) versus a mean defect fill of 2.2 +/- 1.4 mm (50.67%) for defects treated with ABM (P<0.05). Other hard tissue findings showed similar clinically superior results with the use of ABM/P-15. Relative defect fill results showed 81% positive (50% to 100% defect fill) responses with ABM/P-15 and 67% positive responses with ABM. There were 3.5 times as many optimal results (> or = 90% defect fill) with ABM/P-15 and twice as many failures (minimal response) with ABM. Soft tissue findings showed no significant differences between treatments. CONCLUSIONS: These results suggest that the use of the P-15 synthetic cell-binding peptide combined with ABM yields better clinical results than the ABM alone in intrabony periodontal defects.
Adult↗
Correlation of an osteoclast antigen and ruffled border on giant cells formed in response to resorbable substrates.
The osteoclast is the specialized multinucleated cell primarily responsible for the degradation of the organic and inorganic components of bone matrix. The functional and developmental relationship between osteoclasts and foreign body giant cells is unclear. The osteoclast plasma membrane ruffled border juxtaposed to the bone surface is a unique morphologic characteristic of active osteoclasts. In the studies reported here giant cell formation was induced in response to a variety of materials implanted onto the richly vascularized chick chorioallantoic membrane. Light and electron microscopic techniques were used to examine the morphologic characteristics of the giant cells. In addition, immunohistochemical methods were used to demonstrate the appearance of a 150 kD cell surface antigen on chicken osteoclasts recognized by monoclonal antibody 121F. Giant cells that formed in response to mineralized bone particles exhibited ruffled borders and stained positively with the 121F antibody. Many giant cells that formed in response to hydroxyapatite possessed ruffled borders similar to but not as extensive as those observed on giant cells formed on bone. Immunohistochemical localization of the 121F antigen on these cells suggested that the antigen was present, but staining intensity was reduced compared to that of bone-associated giant cells. The formation of mineral matrix complexes by the adsorption to hydroxyapatite of bone extract or osteocalcin enhanced ruffled borders and the presence of the 121F antigen on elicited giant cells. In contrast, giant cells that formed on non-resorbable materials, such as Sepharose beads, mica, and methacrylate, lacked ruffled borders and were negative for the 121F antigen. It appears that expression of the 121F osteoclast antigen correlates with the appearance and extent of ruffled membranes on giant cells. Furthermore, it appears that giant cell ruffled membrane development and the presence of the 121F osteoclast antigen are related to giant cell formation in response to resorbable materials that are subject to extracellular dissolution. Expression of this antigen may be indicative of the developmental and/or functional state of giant cells (osteoclasts) that form on resorbable substrates. In addition, components of the bone matrix, including osteocalcin, in association with bone mineral, lead to elevated levels of this osteoclast antigen.
Allantois↗