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PubMed1982

Evaluation of ossification in jaw defects by radio-nuclide imaging.

The uptake of the bone-seeking radiopharmaceutical 99mTc-MDP in defects of the mandibula was quantitated from scintigraphic images. Activity ratios were calculated by means of ROI-technique. The four selected regions contained two control regions and two with mandibula defects. One defect was filled with a fibrin bonding agent and one remained unfilled. The time course of the ratios showed a significant higher uptake of 99mTc-MDP in the region of the filled defect in comparison with the unfilled defect. The measurement of the residual defect areas from histological sections prepared 18 weeks after introduction of the defects confirmed the much more intensive ossification in the filled defects which could also be derived from the evaluation of the scintigrams. The results indicate that it is possible to follow up osseous repair by quantitative evaluation of scintigraphic images.

Animals↗
PubMed1982

The localization of thiamine pyrophosphatase activity in Meckel's cartilage cells during endochondral ossification.

The cytochemical distribution of thiamine pyrophosphatase (TPPase) activity in Meckel's cartilage cells of the mouse embryo has been studied during the endochondral ossification. All the cartilage cells contain reaction product within the Golgi apparatus. In immature chondrocytes, at the reserve cell zone, TPPase activity is restricted to several inner cisternae of independent Golgi apparatus. In mature cells at the proliferative cell zone, several Golgi complexes form a Golgi network connecting with each other by the TPPase positive tubular stalks. Golgi cisternae, condensing vacuoles and vesicles also contain reaction product. In the hypertrophic chondrocytes located in the calcifying zone, their disorganized Golgi apparatus still retain reaction product. Some chondrocytes, even those located within calcified or opened lacunae, exhibit intact structures and normal cytochemical enzyme distribution. These data indicate the possibility that some chondrocytes may survive and contribute the formation of mandible.

Animals↗
PubMed1995

Characterization of the phosphatidylinositol-specific phospholipase C-released form of rat osseous plate alkaline phosphatase and its possible significance on endochondral ossification.

Alkaline phosphatase activity was released up to 100% from the membrane by incubating the rat osseous plate membrane-bound enzyme with phosphatidylinositol-specific phospholipase C. The molecular weight of the released enzyme was 145,000 on Sephacryl S-300 gel filtration and 66,000 on PAGE-SDS, suggesting a dimeric structure. Solubilization of the membrane-bound enzyme with phospholipase C did not destroy its ability to hydrolyse PNPP, ATP and pyrophosphate. The hydrolysis of ATP and PNPP by phosphatidylinositol-specific phospholipase C-released enzyme exhibited 'Michaelian' kinetics with K0.5 = 70 and 979 microM, respectively. For pyrophosphate, K0.5 was 128 microM and site-site interactions were observed (n = 1.4). Magnesium ions were stimulatory (K0.5 = 1.5 mM) and zinc ions were a powerful noncompetitive inhibitor (Ki = 6.2 microM) of phosphatidylinositol-specific phospholipase C-released enzyme. Phosphatidylinositol-specific phospholipase C-released alkaline phosphatase was relatively stable at 40 degrees C. However, with increasing temperature from 40-60 degrees C, the enzyme was inactivated rapidly following first order kinetics and thermal inactivation constants varied from 5.08 x 10(-4) min-1 to 0.684 min-1. Treatment of phosphatydilinositol-specific phospholipase C-released alkaline phosphatase with Chellex 100 depleted to 5% its original PNPPase activity. Magnesium (K0.5 = 29.5 microM), manganese (K0.5 = 5 microM) and cobalt ions (K0.5 = 10.1 microM) restored the activity of Chelex-treated enzyme, demonstrating its metalloenzyme nature. The stimulation of Chelex-treated enzyme by calcium ions (K0.5 = 653 microM) was less effective (only 26%) and occurred with site-site interactions (n = 0.7). Zinc ions had no stimulatory effects. The possibility that the soluble form of the enzyme, detected during endochondral ossification, would arise by the hydrolysis of the Pl-anchored form of osseous plate alkaline phosphatase is discussed.

Alkaline Phosphatase↗
PubMed1990

Intramembranaceous ossification analyses by a proton microprobe.

The proton induced X-ray emission method in combination with a proton microprobe was applied to study the intramembranaceous ossification. As material sections of mouse embryo skulls from the 17th and 19th day of gestation were used. The morphology of the sample was examined by routine histochemical procedure performed on the sections adjacent to that irradiated by the proton microprobe. The measurements were made in line scan and raster scan mode. The concentrations of P, S, Cl, K, Ca, Fe and Zn were determined at each irradiated point. The average element concentrations were calculated for four parts of each section (bone, cartilage, mesenchymal tissue close to the bone and mesenchymal tissue in other places). The distributions of Ca and P (less markedly than Ca) concentrations almost exclusively correlate with localization of the bone while S, Cl and K concentrations show preference to the cartilage. The amount of inorganic material in flat bones of the 17-day embryo amounts to 14% of the dry mass. The material is characterized by a Ca/P ratio of about 1.6. In the embryo 2 days older the amount of the inorganic phase is practically the same (15%) while the Ca/P ratio approaches 2. This suggests the presence of the precursor phase in the flat bone calcification. It is possible that octacalcium phosphate (Ca/P ratio equals to 1.72) is formed at the onset of the flat bone mineralization which transforms rapidly (in 2 days) to a more stable mineral (defective hydroxyapatite).

Animals↗
PubMed1986

Ossification centers of the pediatric elbow: a rare normal variant.

A 7-year 9-month-old boy sustained a transcondylar humeral fracture during a fall. Radiographs revealed a trochlear ossification center without evidence of a medial epicondylar center. This combination, although apparently quite rare, should be recognized as a normal variant.

Child↗
PubMed1988

A monoclonal antibody against dentin phosphophoryn recognizes a bone protein(s) appearing at the beginning of ossification.

Decalcified and nondecalcified sections of fetal bovine tibia were stained immunohistochemically with a monoclonal antibody against dentin phosphophoryn. In the epiphyseal portion of the long bone, osteoblasts, osteocytes and the bone matrix were stained, but chondrocytes and the cartilage matrix were not. Similar staining was observed in the epiphyseal and diaphyseal portions of bones. These findings suggest that a protein(s) with the same epitope as phosphophoryn may be synthesized and secreted by osteoblasts at the beginning of ossification and may be involved in mineralization of bone tissue. On Western blots of proteins extracted from fetal bovine bone, the antibody reacted with two bands of molecular weights of about 71,000 and 63,000. These proteins and antibody(s) to the proteins may be useful for detection of the phenotype of osteogenesis.

Animals↗
PubMed1995

Differentiation and mineralization in chick chondrocytes maintained in a high cell density culture: a model for endochondral ossification.

Chondrocytes isolated from the proliferative and differentiating zones of 3-wk-old chick growth plates were cultured in the presence of 10% fetal bovine serum (FBS) and ascorbic acid for up to 21 d in a high cell density culture within Eppendorf tubes. The proliferative, differentiating, and calcification properties of the chondrocytes were examined by immunolocalization and by enzyme histochemical and biochemical methods. The cells maintained a chondrocyte phenotype throughout culture: they were round in shape and synthesized both collagen type II and proteoglycans. The expression of a hypertrophic phenotype was evident by Day 3 of culture and from this time onwards characteristics of terminal differentiation were observed. The cells were positive for both alkaline phosphatase (ALP) activity and c-myc protein and the surrounding matrix stained strongly for collagen type X. Small foci of mineralization associated with individual chondrocytes were first evident by Day 6 and more widespread areas of mineralization occupying large areas of matrix were present by Day 15. Mineralization occurred without the addition of exogenous phosphate to the medium. This culture system displays characteristics that are similar in both morphological and developmental terms to that of chick chondrocyte differentiation and calcification in vivo and therefore offers an excellent in vitro model for endochondral ossification.

Alkaline Phosphatase↗
PubMed1996

Studies on the ossification of compound transplantation of porous TCP ceramics and bone marrow.

Following the culture of human red marrow with porous TCP ceramics in vitro, a large amount culturing marrow cells adhered well to the surface and inner wall of TCP ceramics and spread well over the dish bottom 4 weeks after cultivation. After the compound transplantation of porous TCP ceramics with red marrow into quadriceps femoris muscle of rabbits, a huge amount of new bone and marrow formed in these sites on and in the ceramics 4 weeks after transplantation. These results showed an excellent biocompatibility between TCP ceramics and marrow and an excellent osteogenic effect in the compound transplantation. The mechanism of ossification and clinical significance about the compound transplantation were discussed.

Animals↗
PubMed1998

Chondrodiatasis in a patient with spondyloepimetaphyseal dysplasia using the Ilizarov technique: successful correction of an angular deformity with ensuing ossification of a large metaphyseal lesion. A case report.

Distraction through the physis (chondrodiatasis) is a controversial technique with unpredictable results. However, it has been used in the past for the lengthening and correction of angular deformities of long bones. We report the case of an 11-year-old patient with spondyloepimetaphyseal dysplasia (SEMD) who presented with a severe recurvatum deformity of the left proximal tibia secondary to collapse of the tibial plateau into a large metaphyseal cystic lesion. Using the chondrodiatasis technique with a percutaneously applied Ilizarov circular frame, we were able to correct this deformity. Surprisingly, healing and ossification of the metaphyseal lesion was simultaneously observed at the end of the treatment, a finding which, to the best of our knowledge, has not been previously reported.

Bone Cysts↗
PubMed2005

Ossification of the external ear: a case report and review of the literature.

A rare case of idiopathic bilateral ossification of the auricular cartilage and cartilage of the external ear canal is reported. The hardening and thickening of the external ear canals with subsequent narrowing and impaired exposure resulted in symptomatic hearing loss because of the recurrent impediment of inspissated cerumen, the removal of which finally became impossible. Surgical resection of the cartilage external ear canal and tragus was indicated to provide a more permanent treatment of conductive hearing loss. Histological examination revealed complete replacement of the fibroelastic cartilage by normal bone structures showing Haversian canals and bone marrow. The present case report is unique because of the involvement of both the auricles and external ear canals, the symptomatic hearing loss and the surgical treatment to relieve symptoms.

Audiometry↗
PubMed2005

Involvement of cyclic guanosine monophosphate-dependent protein kinase II in chondrocyte hypertrophy during endochondral ossification.

During vertebrate skeletal development, the appendicular skeleton forms through endochondral ossification, which involves the intricately regulated multistep differentiation of mesenchymal cells. During this process, mesenchymal condensations initially differentiate into chondrocytes. Then chondrocytes in the center further differentiate into hypertrophic chondrocytes. Hypertrophic chondrocytes express a number of osteogenic factors and induce bone formation. Although numerous studies have provided novel insights into the regulation and function of cartilage development, little is known about the intracellular signaling pathways regulating chondrocyte hypertrophy. Recent study revealed that cyclic guanosine monophosphate (cGMP)-dependent protein kinase II (cGKII) coupled the stop of proliferation and the start of hypertrophic differentiation of chondrocytes. Herein, we review the molecular mechanism of regulation of chondrocyte hypertrophy by cGKII and the interaction between cGKII and other signaling pathways.

Journal Article↗
PubMed1988

Sonographic assessment of the distal femoral and proximal tibial ossification centers in the prediction of pulmonic maturity in normal women and women with diabetes.

A noninvasive method for the assessment of fetal pulmonic maturity via the sonographic characterization of fetal distal femoral and proximal tibial epiphyseal ossification centers was evaluated. The study population included 82 patients in the third trimester of pregnancy, 47 of whom had uncomplicated gestations, while 35 had insulin-dependent diabetes. In uncomplicated pregnancies a distal femoral epiphysis measuring greater than or equal to 3 mm in the axial plane or the sonographic identification of a proximal tibial epiphysis of any dimension correlated with pulmonic maturity. The combination of a distal femoral epiphysis greater than or equal to 3 mm and the presence of a proximal tibial epiphysis was highly correlated with a lecithin/sphingomyelin ratio greater than or equal to 2:1 (sensitivity 100%, specificity 63%, negative predictive value 100%, and false-negative rate 0%). The Kappa value (K) for testing the agreement between ultrasound results and amniocentesis results was 0.39 (p less than 0.002). In diabetic pregnancies neither a distal femoral epiphysis greater than or equal to 3 mm nor a proximal tibial epiphysis of any dimension correlated precisely with the presence of phosphatidyl glycerol in the amniotic fluid. However, the combination of a distal femoral epiphysis greater than or equal to 3 mm and a proximal tibial epiphysis greater than or equal to 2 mm improved the prediction of phosphatidyl glycerol (sensitivity 83%, specificity 79%, negative predictive value 96%, false-negative rate 17%). The K value in this instance was 0.36 (p greater than or equal to 0.19). It is concluded that the sonographic assessment of the distal femoral and proximal tibial epiphyses and their dimensions in the axial plane may be useful as markers for pulmonic maturity in uncomplicated pregnancies.

Epiphyses↗
PubMed1988

Ultrasonographic assessment of gestational age with the distal femoral and proximal tibial ossification centers in the third trimester.

A prospective sonographic evaluation of the distal femoral and proximal tibial epiphyseal ossification centers in 228 normal pregnant women was carried out from 28 to 40 weeks' gestation. The mean gestational age at which the distal femoral epiphysis and proximal tibial epiphysis were imaged was 34 and 38 weeks, respectively. The distal femoral epiphysis was not identifiable before 28 weeks but was observed in 72% of fetuses at 33 weeks and in 94% of fetuses at 34 weeks' gestation. The presence of a distal femoral epiphysis measuring 1 or 2 mm was associated with a gestational age of greater than 33 weeks in 87.0% of fetuses, whereas a distal femoral epiphysis measuring greater than or equal to 3 mm was associated with a gestational age greater than 37 weeks in 85% of fetuses. The proximal tibial epiphysis, which was absent before 34 weeks' gestation, was observed in 56% of fetuses at 36 weeks, in 80% of fetuses at 37 weeks, and in 100% of fetuses at 39 weeks of gestation. The presence of a proximal tibial epiphysis of 1 or 2 mm was associated with a gestational age of greater than 36 weeks in 88% of fetuses, whereas a proximal tibial epiphysis greater than or equal to 3 mm was associated with a gestational age of greater than 38 weeks in 94% of fetuses. The sonographic evaluation of distal femoral epiphysis/proximal tibial epiphysis can be used as independent markers for estimation of gestational age during the third trimester, a period in which standard fetal biometric estimates of gestational age are least accurate.

Epiphyses↗
PubMed1993

Genital and nongenital teratogenesis of prenatal progestogen therapy: the effects of 17 alpha-hydroxyprogesterone caproate on embryonic and fetal development and endochondral ossification in the C57B1/6J mouse.

OBJECTIVE: The current study assesses the effects of 17 alpha-hydroxyprogesterone caproate on embryo-fetal development, with a particular focus placed on intrauterine endochondral bone development and growth. STUDY DESIGN: Primigravid C57 BI/6J mice were administered 17 alpha-hydroxyprogesterone caproate by means of subdermal pellets designed to deliver doses of 0.5, 5.0, and 50.0 mg/kg/day on gestational days 7 through 19. Assuming a dosage regimen of 250 mg per 50 kg per week in humans for the treatment of threatened spontaneous abortion, the doses used were 0.7, 7.0, and 70.0 times the human dose equivalent. The time course of exposure encompassed the period of organogenesis through the late fetal period. RESULTS: No maternal toxicity was noted throughout the course of 17 alpha-hydroxyprogesterone caproate administration. Fetal weight at term and the percentage of resorptions, dead fetuses, male fetuses, and malformations at term were not significantly affected at any evaluated dose when compared with control group values. The steroid did not exert a significant influence on any assessed parameter of endochondral bone development or growth, even at a dosage concentration that far exceeded the human therapeutic dose equivalent. As would be expected, fetal weight was directly correlated with diaphyseal length and also influenced the relationship of cartilage to osseous tissue in both the humeri and femora. There were no 17 alpha-hydroxyprogesterone caproate dose and fetal sexual phenotype interactive effects noted with regard to any developmental or bone growth parameter measured, thus discounting any sexual phenotype-related toxic or teratogenic sensitivities. CONCLUSION: The data indicate that 17 alpha-hydroxyprogesterone caproate administered at doses of up to 70 times the human dose equivalent did not induce increases in the frequency of genital and, importantly, nongenital teratogenesis in spite of exposure during the critical period of organogenesis. Additionally, the steroid did not affect limb morphogenesis or endochondral ossification.

17 alpha-Hydroxyprogesterone Caproate↗
PubMed1996

Excision of heterotopic ossification around the knee following brain injury.

We present the results of five patients who underwent resection of heterotopic ossification around the knee following traumatic brain injury. All the patients had improved function and no pain at a minimum of 1 year follow up, irrespective of their preoperative neurological status. Our philosophy for such surgery is outlined and, although the surgical goals may seem small, the outcome for any individual patient may be considerable.

Adult↗
PubMed1995

Effect of therapeutic ultrasound on endochondral ossification.

The effect of therapeutic doses of ultrasound was tested on endochondral ossification of in vitro developing metatarsal long bone rudiments of 16- and 17-day-old fetal mice. Bone growth, calcification and resorption following exposure to several doses of pulse-wave (PW) or continuous-wave (CW) ultrasound were examined. PW was applied at intensities between 0.1 W cm-2 and 0.77 W cm-2 (Isatp) and CW intensities were 0.1 W cm-2 or 0.5 W cm-2 (Isata). After 1 week of culture, the metatarsal long bone rudiments were fixed and paraffin sections were prepared for histological evaluation and for measurement of the relative contribution of the various cartilage zones to the total bone length. In contrast to treatment with CW ultrasound, treatment of 16-day-old metatarsal long bone rudiments with PW ultrasound resulted after 4 days of culture in significantly increased longitudinal growth. Histology revealed a significant increased length of the proliferative zone, whereas the length of the hypertrophic cartilage zone was unaltered. This might indicate that proliferation of the cartilage cells is stimulated without influence on cell differentiation.

Animals↗
PubMed2006

Erythropoietin (EPO): EPO-receptor signaling improves early endochondral ossification and mechanical strength in fracture healing.

Beyond its role in the regulation of red blood cell proliferation, the glycoprotein erythropoietin (EPO) has been shown to promote cell regeneration and angiogenesis in a variety of different tissues. In addition, EPO has been indicated to share significant functional and structural homologies with the vascular endothelial growth factor (VEGF), a cytokine essential in the process of fracture healing. However, there is complete lack of information on the action of EPO in bone repair and fracture healing. Therefore, we investigated the effect of EPO treatment on bone healing in a murine closed femur fracture model using radiological, histomorphometric, immunohistochemical, biomechanical and protein biochemical analysis. Thirty-six SKH1-hr mice were treated with daily i.p. injections of 5000 U/kg EPO from day 1 before fracture until day 4 after fracture. Controls received equivalent amounts of the vehicle. After 2 weeks of fracture healing, we could demonstrate expression of the EPO-receptor (EPOR) in terminally differentiating chondrocytes within the callus. At this time point EPO-treated animals showed a higher torsional stiffness (biomechanical analysis: 39.6+/-19.4% of the contralateral unfractured femur) and an increased callus density (X-ray analysis (callus density/spongiosa density): 110.5+/-7.1%) when compared to vehicle-treated controls (14.3+/-8.2% and 105.9+/-6.6%; p<0.05). Accordingly, the histomorphometric examination revealed an increased fraction of mineralized bone and osteoid (33.0+/-3.0% versus 28.5+/-3.6%; p<0.05). Of interest, this early effect of the initial 6-day EPO treatment had vanished at 5 weeks after fracture. We conclude that EPO-EPOR signaling is involved in the process of early endochondral ossification, enhancing the transition of soft callus to hard callus.

Animals↗