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[Effects of oral administration of various non-steroidal anti-inflammatory drugs on bone growth and bone wound healing in mice].

Morphometric and microdensitometric studies on normal bone growth and bone wound healing in growing ICR strain male mice were carried out as preliminary investigations. Then the effects of oral administration of non-steroidal anti-inflammatory drugs on bone growth and bone wound healing were studied. Femur length and width, and bone mineral contents in the femur as judged by microdensitometry of the soft x-ray films of the cortical portion of the femur showed time-dependent linear increase between 3 and 6 weeks of age. Wound holes were made in the parietal bones of mice with a dental bar at 4 weeks of age, and uncalcified area in the holes was determined by image analysis of soft x-ray films. The area decreased their size with the advance in age, especially a marked decrease was observed 2 weeks after the operation. From the above results, 4 week-old male mice were employed and 8 kinds of non-steroidal anti-inflammatory drugs (indomethacin, 1 mg/kg; mefenamic acid, 10 mg/kg; flufenamic acid, 10 mg/kg; diclofenac sodium, 1 mg/kg; ibuprofen, 10 mg/kg; naproxen, 10 mg/kg; phenylbutazone, 10 mg/kg and aspirin, 50 mg/kg) and dexamethasone (1 mg/kg), a steroidal anti-inflammatory drugs were administered orally every day for 9 or 11 days except Sunday, and 15 days after the initial administration, the length and width of the femur were measured. The bone mineral contents were determined by microdensitometry of the soft x-ray film of the femur. Hydroxyproline contents of the femur were also determined. Wound holes were made in the parietal bones at 4 week-old and administered the drugs from one day before the operation for 9 days and the uncalcified area in the wound hole was determined 2 weeks after the operation with an image analyzer. The results obtained were as follows: 1. A significant inhibition of the growth in length of the femur was observed by dexamethasone, diclofenac sodium and naproxen administration for 9 and 11 days. A significant inhibition of the growth in width was also observed by dexamethasone and diclofenac sodium administration for 9 days and by dexamethasone administration for 11 days.(ABSTRACT TRUNCATED AT 400 WORDS)

Administration, Oral↗

Infantile myofibromatosis. Report of two cases.

Two cases of infantile myofibromatosis were presented. Case 1 was a 6-month-old girl with multiple nodular lesions in the left parietal bone, dermis of abdominal wall, chest wall, right upper arm, and bilateral femur. Case 2 was a 12-month-old girl with solitary nodular lesions in the left parietal bone having a tendency to enlarge. Histologically, these tumorous lesions consisted of broad bundles of well oriented spindle-shaped cells superficially resembling smooth muscle tissue. In electron microscopy, the constituent cells combined ultrastructural characteristics of both fibroblasts and smooth muscle cells., Immunoperoxidase method showed that anti-smooth muscle antibodies binded to the cytoplasm of the bundle forming cells. From these results, it may be concluded that the lesion consisted of pure proliferation of myofibroblasts, and its peculiar growth behavior was discussed, though the histogenesis still remains obscure.

Female↗

The skull and mandible of the African elephant (Loxodonta africana).

In the present study the bones of the skull, excluding the hyoid apparatus, are described. All the bones are aerated by sinuses. In the occipital bone the squamous part is aerated from the sinus of the parietal bone, the lateral part is aerated from the tympanic bulla and the basal part from the sinus of the basisphenoid bone. Condylar foramens and hypoglossal canals are absent. A small interparietal bone is present at birth. At an early age it fuses with the surrounding cranial bones. The squamous part of the temporal bone lies sagittally in young animals, but moves progressively to a transverse plane as the animals age. A foramen lacerum is represented by jugular and oval foramens and the carotid canal. The body of the basisphenoid bone is excavated by the massive maxillary tuberosity. The latter extends to the oval foramen and contains the developing molar teeth. The ethmoturbinate, nasal and lacrimal bones are exceptionally small. In old bulls the palatine process of the incisive bones and their sinuses are gradually displaced by the palatine process of the maxillae.

Animals↗

Immunogold localization of beta 1-integrin in bone: effect of glucocorticoids and insulin-like growth factor I on integrins and osteocyte formation.

Insulin-like growth factor I (IGF-I) and high-dose glucocorticoids exert opposite effects on bone formation. Because integrins are involved in cell and matrix organization, the effect of glucocorticoids and IGF-I on integrins was investigated in bone. An immunogold transmission electron microscopic (TEM) method was developed and applied to an organ culture system of 20-day fetal rat parietal bones, which mineralize in vitro. In parietal bone culture, 100 mM corticosterone treatment for 72 hr decreased calcification by 29%, disrupted osteoblast organization, and decreased the number of osteocytes. The quantity of osteoblast processes and the number of osteocytes per unit bone area were decreased by 48% and 56%, respectively. This effect was dose-dependent. The beta 1-integrin subunit was localized equally to apical and basal osteoblast surfaces by immunogold TEM. Compared to untreated control cultures, corticosterone (100 nM) decreased beta 1 by one third. In contrast, treatment with IGF-I for 72 hr increased calcification by 38%, cell processes by 71%, and osteocytes per unit area of bone by 107%. The number of gold particles localizing beta 1 on the osteoblast plasma membrane doubled, almost entirely on the apical surface of the osteoblast. Glucocorticoids and IGF-I had no significant effect on beta 1 levels in osteocytes. In conclusion, glucocorticoids and IGF-I modulate integrin levels on osteoblasts, and influence osteocyte formation and bone calcification. However, neither glucocorticoids nor IGF-I alter beta 1-integrin levels on osteocytes.

Animals↗

[Study on renal osteodystrophy using 2-compartment model analysis of bone scintigraphy. 1. Clinical significance of K Index].

The dynamic analyses of bone scintigraphy were performed in 30 cases of hemodialysed patients. The regression analyses between biochemical data or duration and K indexes obtained from the 2-compartment model analysis in the maxillofacial region were carried out. The obtained results were as follows. 1) The duration and K indexes were significantly correlated with 1 or 2% of significance level in the parietal bone and the zygomatic bone. 2) Serum Ca and K indexes were highly correlated in every settled ROI. The correlation coefficients were distributed from 0.58 to 0.79. In the cranial bone the correlation were more higher than in maxilla and mandibular bone. 3) Serum ALP and K indexes have more higher significant level of correlation than the results of Ca. The correlation coefficients were distributed from 0.48 to 0.83. The level of significance were changed by location. The highest correlation coefficient was 0.83 in the parietal bone. 4) Serum c-PTH and K indexes have significant correlation in every settled ROI with the correlation coefficient from 0.59 to 0.69. The significance level were nearly equal in every location. 5) In the case of subtotal parathyroidoectomy the K indexes which were abnormally high in preoperation showed the tendancy to decrease toward normal range in postoperation. These results indicated that K indexes reflected the bone change caused by renal osteodystrophy. And the K indexes considered to be usefull to estimate the bone improvement.

Bone and Bones↗

Bacterial exposure required to induce rabbit calvarial bone graft infection by superficial contamination.

Little is known about the minimum bacterial dose required to induce infections by superficial contamination alone (i.e., not direct inoculation). This study quantified superficial rabbit calvarial bone graft contamination by exposing parietal bone grafts to Staphylococcus aureus. Two 5 x 5-mm full-thickness calvarial bone grafts were harvested from 40 rabbits and contaminated with different concentrations (range = 10(7)-10(11) colony-forming units [CFUs]/mL) of S. aureus by immersion. One graft from each rabbit was cultured immediately, and the other was inset into a calvarial donor site, fixed in place with wire, and observed for evidence of infection. At harvest (28 days postoperatively), a bacterial exposure of greater than 10(8) CFUs/mL of S. aureus was required to induce infections in the rabbit calvarial grafts (p < 0.001, Student's t-test), transmitting an infectious dose of approximately more than 10(4) CFUs per graft. These results validate the previously known dogma regarding the number of organisms needed to manifest infection [1,2] and highlight a multifold increment in bacterial count needed to transmit this infectious dose by superficial contamination alone. Such a model may be useful in investigating the efficacy of various treatment modalities of contaminated bone.

Air Microbiology↗

Scanning electron microscopic and light microscopic observations on morphological changes of freeze-dried bone implantation in rats: comparison with fresh autogenous bone transplantation.

Bone remodelling after the implantation of freeze-dried autogenous bone in rat parietal bone was compared with fresh autogenous bone transplantation, using a scanning electron and light microscope revealed the time intervals after transplantation/implantation. The light microscope revealed the time delay of the bone remodelling in the implantation, compared with the transplantations. The scanning electron microscope showed that the differences between the two groups were in the states of bone union and bone resorption. In the fresh bone group, the newly-formed bone filled the spaces between host and the transplanted bones at 2 to 3 weeks after the transplantation: the newly-formed bone fused and melted into the transplanted bone. New bone formation was more dominant on the bone surface in the dura mater side than in the skin side. The union was almost completed at 5 weeks. In freeze-dried bone implantation, the bone union in the contact space was very poor and the implanted bone was mainly covered by the new bone, which developed from the host bone surface in the dura mater side at 2 to 3 weeks after the implantation. What is noteworthy is that bone resorbed areas showing numerous Howship's lacunae were mainly observed on the host bone surface in the vicinity of newly-formed bone. However in freeze-dried bone implantation, the bone resorption was greater on the host and implanted bone surface than that of fresh bone transplantation: the resorption of host bone was considerably larger at certain periods after freeze-dried bone implantation. The present results show that the healing process of freeze-dried bone implantation, even though autogenous bone was used, differed from that of fresh autogenous bone transplantation, and the differences are concerned not only with time sequences but also with qualitative changes. This suggests that the host would have some different responses to the freeze-dried autogenous bone from fresh materials.

Animals↗

Impact of nicotine on bone healing.

A limited number of experimental animal studies and in vitro data confirm that nicotine impairs bone healing, diminishes osteoblast function, causes autogenous bone graft morbidity, and decreases graft biomechanical properties. Therefore, our long-term goal is to develop an effective therapy to reverse the adverse impact of nicotine from tobacco products. However, before accomplishing this goal, we had to develop an animal model. Our hypotheses were nicotine administration preceding and following autogenous bone grafting adversely affected autograft incorporation and depressed donor site healing in a characterized animal wound model. Hypothesis testing was accomplished in bilateral, 4-mm diameter parietal bone defects prepared in 60 Long-Evans rats (male, 35-day-old). A 4-mm diameter disk of donor bone was removed from the left parietal bone and placed in the contralateral defect. The donor site served as a spontaneously healing bone wound. The rats were partitioned equally among three doses of nicotine administered orally in the drinking water (12.5, 25, and 50 mg/L). For each dose, the duration and sequence of nicotine treatment followed four courses, including no nicotine and designated combinations of nicotine administration and abatement prior to and following osseous surgery. Experimental sites were recovered on 14 and 28 days postsurgery, responses quantitated, and data analyzed by analysis of variance and post hoc statistics (p < or = 0.05). We developed a convenient and effective osseous model, and the results validated our hypothesis that nicotine negatively impacts on bone healing.

Animals↗

Recovery course of full-thickness skin defects with exposed bone: an evaluation by a quantitative examination of new blood vessels.

BACKGROUND: Full-thickness skin defects with exposed bone are often hard to heal. The lack or delayed re-vascularization is considered one of the major causes, and the periosteum is also suggested to have an important role in tissue regeneration. MATERIALS AND METHODS: Full-thickness skin defect wounds with exposed bone were made in the parietal region of Wister rats. The periosteum of the exposed parietal bone was removed in the periosteum-lacking group, but maintained in the control group (periosteum-intact group). The wound was covered by an artificial dermis made of collagen. The wound healing process was histologically compared. Double immunostaining of alpha-smooth muscle actin (SMA) and von Willebrand factor (vWF) was used for re-vascularization examination, and the blood vessel density in the artificial dermis was quantified. RESULTS: The density of the blood vessels in the uninjured parietal tissue was approximately 80 vessels/mm(2). To reach this density, 7 and 21 days were required for the control (periosteum-intact) and the periosteum-lacking groups, respectively. This coincided with complete revascularization, fibroblast migration and the reentry of blood vessels to the upper layer of the wound were observed. CONCLUSION: The described results support the importance of the periosteum in the full-thickness skin defect healing process.

Animals↗

Temporal osteocutaneous island flaps for complete reconstruction of cleft palate defects.

In each of four patients, closure of a wide cleft palate was performed with a temporal osteocutaneous island flap (TOCI flap) based on the superficial temporal artery. These were fabricated island fasciocutaneous flaps composed of temporoparietalis fascia and galea with pericranium, surfaced with split skin grafts. Two flaps carried with them a segment of vascularized parietal bone (TOCI-I). In the other two flaps, the osseus component was supplied by placing an osteogenic powder in a pericranial pocket: a temporal artery "osteogenic" cutaneous flap (TOCI-II). In each case the flap closed the defect and lengthened the palate without the need for elevation of palatal mucoperiosteum. Growth of the parietal bone segment has been observed in the first patient (the only one to have a follow-up CT scan). This patient, the eldest, is developing normal speech so far. The others are still too young for evaluation.

Cleft Palate↗

Computed measures on computed tomograms of the head.

Measures of average parietal bone mass (APBM) and average parietal skull width (APSW) were developed using automatic image processing techniques and a special polar coordinate system. The precision of the technique is about 2%; the measures are obtained by processing one section of a computed tomographic head scan obtained without special apparatus. In males the APBM does not decrease with age; in females it splits into two groups of age, probably based on postmenopausal changes. The APSW in both males and females increases slightly with age. The clinical utility of the measures is discussed.

Cephalometry↗

Differential expression patterns of Runx2 isoforms in cranial suture morphogenesis.

Runx2 (previously known as Cbfal/Pebp2alphaA/AML3), a key transcription factor in osteoblast differentiation, has at least two different isoforms using alternative promoters, which suggests that the isoforms might be expressed differentially. Haploinsufficiency of the Runx2 gene is associated with cleidocranial dysplasia (CCD), the main phenotype of which is inadequate development of calvaria. In spite of the biological relevance, Runx2 gene expression patterns in developing calvaria has not been explored previously, and toward this aim we developed three probes: pRunx2, which comprises the common coding sequence of Runx2 and hybridizes with all isoforms; pPebp2alphaA, which specifically hybridizes with the isoform transcribed with the proximal promoter; and pOsf2, which hybridizes with the isoform transcribed with the distal promoter. These probes were hybridized with tissue sections of mouse calvaria taken at various time points in development. Runx2 expression was localized to the critical area of cranial suture closure, being found in parietal bones, osteogenic fronts, and sutural mesenchyme. Pebp2alphaA and Osf2 showed tissue-specific expression patterns. The sites of Pebp2alphaA expression were almost identical to that of pRunx2 hybridization but expression was most intense in the sutural mesenchyme, where undifferentiated mesenchymal cells reside. The Osf2 isoform was strongly expressed in the osteogenic fronts, as well as in developing parietal bones, where osteopontin (OP) and osteocalcin (OC) also were expressed. However, in contrast to Pebp2alphaA, Osf2 expression did not occur in sutural mesenchyme. Pebp2alphaA also was expressed prominently in primordial cartilage that is found under the sutural mesenchyme and is not destined to be mineralized. Thus, Osf2 isoforms contribute to events later in osteoblast differentiation whereas the Pebp2alphaA isoform participates in a wide variety of cellular activities ranging from early stages of osteoblast differentiation to the final differentiation of osteoblasts.

Animals↗

Tissue origins and interactions in the mammalian skull vault.

During mammalian evolution, expansion of the cerebral hemispheres was accompanied by expansion of the frontal and parietal bones of the skull vault and deployment of the coronal (fronto-parietal) and sagittal (parietal-parietal) sutures as major growth centres. Using a transgenic mouse with a permanent neural crest cell lineage marker, Wnt1-Cre/R26R, we show that both sutures are formed at a neural crest-mesoderm interface: the frontal bones are neural crest-derived and the parietal bones mesodermal, with a tongue of neural crest between the two parietal bones. By detailed analysis of neural crest migration pathways using X-gal staining, and mesodermal tracing by DiI labelling, we show that the neural crest-mesodermal tissue juxtaposition that later forms the coronal suture is established at E9.5 as the caudal boundary of the frontonasal mesenchyme. As the cerebral hemispheres expand, they extend caudally, passing beneath the neural crest-mesodermal interface within the dermis, carrying with them a layer of neural crest cells that forms their meningeal covering. Exposure of embryos to retinoic acid at E10.0 reduces this meningeal neural crest and inhibits parietal ossification, suggesting that intramembranous ossification of this mesodermal bone requires interaction with neural crest-derived meninges, whereas ossification of the neural crest-derived frontal bone is autonomous. These observations provide new perspectives on skull evolution and on human genetic abnormalities of skull growth and ossification.

Animals↗

Difference in size of bone islands formed by isolated bone cells transplanted intramuscularly under various conditions.

Bone cells isolated from the whole calvaria (2 x 10(6)) from either central or peripheral parts of parietal bones (1 x 10(6)) and from scapulas (2 x 10(6)) were allowed to adhere to devitalized calvarial bones in the number indicated in brackets and transplanted intramuscularly (supported transplants). Whole calvaria bone cells (2.4 or 8 x 10(6) cells per transplant) were also injected intramuscularly as free transplants. Calvarial cells produced solid bone islands with small intraosseous cavities, while bone formed by scapular cells contained large medullary spaces. The size of bone islands formed in transplants and the shortest distance between the neighbor islands were measured. The results of these measurements were similar in all groups of free transplants. The size of bone islands formed in supported transplants of cells from the whole calvaria or from central and peripheral parts of parietal bones was also roughly similar, but the shortest distance between islands was larger than in the free transplants. Furthermore, in these groups of transplants bone islands considerably larger than the largest islands in free transplants were present. Scapular bone cells formed islands much larger than those produced by calvarial cells. Bone islands formed by calvarial cells in free transplants were separated by bands of fibrous tissue which was absent in supported transplants. It appears that this tissue could limit growth and/or fusion of neighbor bone islands and in this manner influence their size. The population of transplanted scapular cells contained numerous stromal elements which could form an exclusion area inaccessible to local cells from the site of transplantation and thus favour formation of large bone islands within this area.

Animals↗

Bone induction using autogenous bone mixed with demineralised bone matrices.

The aim of the present study was to examine the healing of endochondral (EC) autogenous bone grafts in the presence of demineralised bone matrix prepared from intramembranous bone (DBMIM), or prepared from endochondral bone (DBMEC) using quantitative analysis. Thirty bone defects were created on the parietal bone of fifteen New Zealand White rabbits. In the experimental groups, five defects were grafted with EC bone, five defects were grafted with EC bone mixed with DBMIM (EC-DBMIM) and six defects were grafted with EC bone mixed with DBMEC (EC-DBMEC). In the control groups, ten defects were left empty (passive control) and four defects were grafted with rabbit skin collagen (positive control). They were all sacrificed at day fourteen post grafting, and the defects were prepared for histological analysis. Serial sections were cut across the whole defect. Quantitative analysis was performed on 152 sections of the experimental groups by image analysis. Four hundred and fourteen per cent more new bone was formed in defects grafted with composite EC-DBMIM than those grafted with EC bone alone (p < 0.001). Eighty-five per cent more new bone was formed in defects grafted with composite EC-DBMEC than those grafted with EC bone alone (p < 0.001). No bone was formed in either passive or positive controls. In conclusion, DBM, especially DBMIM, have extremely high osteoinductive properties and greatly enhance the integration of EC bone grafts with defects created in IM bone.

Analysis of Variance↗

[Double contours and companion shadows in the skull associated with anaemias (author's transl)].

Lateral X-ray pictures of the skull in certain several and chronic disorders of the blood as thalassaemia, congenital haemolytic anaemia, sickle cell anaemia and iron deficiency disease show frequently changes of the outlines of the cranial bones. They consist of a double contour of the outer cranial border and on the interior side below the sagittal sutura of a band-like shadow or of multiple stripes and lines running parallel to the carnial vault. They concern the parietal bones and may extend from the bregma till to the lamda or occupy only a part of this distance. The roentgenological features are due to the enlargement of the crainal bones, the bulging out of both parietal bondes, the sometimes enlarged and deepened borders of the sulcus sagittalis superior and particularly to the furrow and depression on the skull above the sagittal suture caused by the protruding parietal bones on both sides. As these different abnormal structures must be passed by the picturing X-ray, effects of superposition and interference are produced. Longitudinal ridges or bony edges which could explain the roentgenological findings could not be established. Since the peculiar alterations of the cranial bones are mainly found in the mentioned blood disorders, where they are caused by the overgrowing red marrow, they also display the same roentgenological features. These features are, therefore, a characteristic sign of these diseases.

Adolescent↗

The thickness of the skull in Korean adults.

New instrumentation and techniques for skeletal fixation have been developed in response to concerns regarding variability of the skull base and the calvaria. The fear of intracranial penetration has limited the use of internal or external fixation to the skull base and the calvaria. Despite potential clinical usefulness, limited anthropometric data have been reported on the skull base and the calvaria. The purpose of this study was to measure the average thickness of various points of the skull base and the calvaria of Korean adults. Fifty-one Korean adult skull bases and calvaria, which were cut 2 cm above the supraorbital margin anteriorly and 2 cm above the inion posteriorly, were measured in this study. Thickness of 13 clinically important points were measured with a Teclock GM-21 Calipergauge (Teclock Co., Tokyo, Japan). The mean thickness of the two opposite sides were compared using paired t-tests. They did not differ significantly from the others except for three points. Frontal bone A (located 1 cm above the supraorbital margin and 1.5 cm lateral to the midline) was 6.35 +/- 2.88 mm, frontal bone B (located at the midpoint of the frontal midpoint and the frontoparietal suture on the cut edge) was 5.24 +/- 1.50 mm, frontal bone C (located at the midpoint of the vertex and frontal bone B) was 6.63 +/- 1.77 mm, the pterion was 3.19 +/- 0.85 mm, the midpoint of the squamous temporal was 1.96 +/- 0.65 mm, parietal bone A (located 1.5 cm below the middle meningeal groove) was 3.99 +/- 1.11 mm; parietal bones B and C (divided at the distances between the frontoparietal suture and the occipital midpoint with a ratio of 1:3 and 2:3 respectively on the cut edge) were 4.71 +/- 1.06 mm and 5.35 +/- 0.91 mm respectively, occipital bone A (located 1.5 cm posterolateral from the foramen magnum) was 2.31 +/- 1.00 mm, occipital bone B (located 1.5 cm lateral to the midline and 0.5 cm above the inion) was 5.41 +/- 1.50 mm, occipital bone C (located occipital to the midpoint and 2 cm above the inion) was 8.2 +/- 1.67 mm, the orbital roof was 1.37 +/- 1.08 mm, and the orbital lateral wall was 2.25 +/- 1.06 mm. The data give practical information for determining the position and depth for safe and effective internal or external fixation during craniofacial procedures.

Adult↗

Cranial vault expansion using transcutaneously activated magnetic implants.

The technique of distraction osteogenesis has not been widely used in the treatment of problems of the head and face because of the need for external fixators. By using magnetic, rather than mechanical, forces to drive bone movement we hope to expand the applications of the technique to include the treatment of cranial vault deformities. Thirty immature rabbits were studied. Twenty-six of the rabbits underwent operations. Each of the 26 had 2 magnets fixed to its skull: one on the left parietal bone and one on the right parietal bone. Incomplete circumferential osteotomies were then performed around each magnet on 13 of the rabbits. A head frame was attached to each animal. Head frames without magnets were placed on 6 of the rabbits (3 with osteotomy, 3 without osteotomy) while 20 of the animals (10 with osteotomy, 10 without osteotomy) had 2 magnets mounted on the frames which were of opposite polarity to those implanted. The remaining 4 rabbits served as nonoperative controls. Six weeks later all of the animals were sacrificed. There were significant differences in the cranial volumes and contours between the groups of animals. Many of the differences were increased by coincident osteotomy. Associated histologic findings are described.

Animals↗