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Biomedical subjects

M Neo

Publications and source records attributed to M Neo.

27 records · Page 2Linked to original sources

Bone-bonding ability of bioactive bone cement under mechanical stress.

Bioactive bone cement (BABC) is able to bond to bone through a Ca-P rich layer. It was evaluated so far in a rat tibial model, where no mechanical stresses are supposed to take place. The objective is to investigate the behavior of BABC in the environment of posterolateral spinal fixation model, in which the bone cement interface is exposed to continuous mechanical stress. Japanese white rabbits were used. Fixation of L5-L6 segment was done by wiring the spinous and transverse processes of L5 and L6 vertebrae. Then BABC was applied over the transverse processes and the intertransverse process membrane on both sides. Polymethylmethacrylate (PMMA) bone cement was used similarly in the control group. Animals were sacrificed after 1 day, 4, 8, and 16 weeks postoperatively. Bone cement interface was examined using Giemsa surface staining and SEM, and affinity index was measured. Biomechanical testing was done nondestructively in right and left torsion. BABC bonded to bone directly with no intervening soft tissue at 4, 8, and 16 weeks, while soft tissue was consistently seen between PMMA bone cement and bone. BABC-spine constructs were stiffer than PMMA-spine constructs at all time intervals. BABC bonded directly to bone under mechanical stress and afforded stiffer fixation than PMMA bone cement.

Animals↗

Osteoblast reaction at the interface between surface-active materials and bone in vivo: a study using in situ hybridization.

Osteoblast activation after implantation of two kinds of surface-active material in bone was investigated chronologically using in situ hybridization with digoxygenin-labeled procollagen alpha 1(I) complementary RNA probe. The bioactive materials used were hydroxyapatite (HA) and apatite- and wollastonite-containing glass-ceramic (A-W GC). A hole was drilled bilaterally in the distal epiphysis of rabbit femurs with subsequent implantation of HA or A-W GC cylinders in a press-fit manner. Specimens were collected at 3, 7, 14, and 28 days after operation and decalcified. Then the undecalcified implant cores were pushed out of the hole without causing damage to the bony side of the interface. In situ hybridization documented no qualitative differences in the expression of procollagen alpha 1(I) RNA between HA and A-W GC. Few osteoblasts at the bone-material interface showed a specific signal at day 3, whereas many osteoblasts were positive around the materials at days 7 and 14, indicative of active new bone formation. The positive osteoblasts seemed to originate from preexisting trabeculae and lined the trabeculae, newly formed bone, and material surface. At day 28, many osteoblasts lining material-surrounding bone were negative, whereas those in remodeling canals were positive, suggesting that the bone was in the remodeling stage after bone formation. These findings were comparable to those with beta-tricalcium phosphate in a previous study, thus suggesting osteoconductive bone formation on HA and A-W GC.

Animals↗

Temporal and spatial patterns of osteoblast activation following implantation of beta-TCP particles into bone.

Temporal and spatial patterns of osteoblast activation around beta-TCP particles implanted into bone were analyzed by in situ hybridization with digoxygenin-labeled procollagen alpha 1(I) RNA probes. beta-TCP particles (150-300 microns in diameter) were implanted into rat tibiae, and specimens were collected 3, 5, 7, 14, and 28 days after operation. Activated osteoblasts displayed intense procollagen alpha 1(I) RNA specific labeling. At day 3, osteoblasts lining pre-existing trabeculae in places showed a specific signal. Additionally, scattered activated cells compatible with preosteoblasts also were observed in the vicinity of the trabeculae among red blood cells that filled the space between beta-TCP particles. Osteoblast activation on the surface of beta-TCP rarely was observed. At days 5 and 7, osteoblast activation and bone formation advanced centripetally. At the forefront of bone formation positive cells were scattered in the blood cell clots, and some of the positive cells colonized forming new bone matrix. Formation of new bone did not always begin at the surface of beta-TCP. At day 14, most of the beta-TCP particles were tightly associated with newly formed bone, and the number of positive osteoblasts was reduced. At day 28, absorption of the newly formed bone and the beta-TCP by multinuclear cells was sporadically demonstrated. Such cells often were accompanied by active osteoblasts, suggesting early bone remodeling. In conclusion, in situ hybridization with procollagen alpha 1(I) was employed to demonstrate precisely the mode of recruitment of bone cell precursors. beta-TCP does not positively guide collagen I expressing bone cells along its surface. It has no apparent effects on bone regeneration.

Animals↗

Analysis of osteoblast activity at biomaterial-bone interfaces by in situ hybridization.

To investigate the effects of bioactive materials on bone formation in vivo, a new experimental model using in situ hybridization has been developed. A hole was drilled bilaterally in the distal epiphysis of rabbit femurs with subsequent implantations of beta-tricalcium phosphate (beta-TCP) cylinders in a press-fit manner. Specimens were collected at 3, 7, 14, and 28 days after operation. Femurs with empty drilling holes, and normal distal femurs without operation were used as controls. All specimens were decalcified and hybridized with a procollagen alpha 1(I) complementary RNA probe labeled with digoxygenin. In normal-bone sections, procollagen alpha 1(I) RNA was clearly demonstrated in periosteal osteoblasts, in osteoblasts in the mineralizing zone adjacent to growth plates, and in osteoblasts lining remodeling canals. As for beta-TCP, labeled osteoblasts around the material were not found at day 3, whereas they were most intensively observed at day 7 and a little less at day 14, in accordance with new-bone formation around the material. Weaker signals were also detected in fibroblasts at day 7. At day 28, osteoblasts lining the surface of newly formed bone were mainly negative, whereas those adjacent to the resorption sites of the beta-TCP showed positive signals, demonstrating an active remodeling at the material surface. The temporal expression of procollagen alpha 1(I) RNA in the beta-TCP specimens was fundamentally the same as that in the empty-hole specimens, suggesting no remarkable acceleration or suppression of bone-forming activity of osteoblasts by beta-TCP, which is consistent with osteoconductive bone formation. This in situ hybridization method was suggested to be a powerful tool in analyzing the biological effects of bioactive materials.

Animals↗

Ultrastructural study of the A-W GC-bone interface after long-term implantation in rat and human bone.

The interface between apatite- and wollastonite-containing glass-ceramic (A-W GC) and bone after long-term implantation was studied by scanning and transmission electron microscopy (SEM and TEM) using rat and human specimens. First, particles of A-W GC (100-220 microns in diameter) were implanted into rat tibiae, and specimens were prepared for observation at 24, 48, 72, and 96 weeks after the operation. These long-term specimens showed an A-W GC-bone interface different from that at an earlier stage, which was investigated in our previous studies. SEM showed that the Ca-P-rich layer was wider, suggesting that leaching of ions from the A-W GC had continued even after bonding with bone. In some regions, the material particles were evidently replaced by the bone. TEM showed that the intervening apatite layer had become indistinct, and that A-W GC had intermingled with bone at the interface. In some regions, the surface of the A-W GC was degraded. These findings suggest that the surface region of A-W GC is slowly replaced by bone. Second, a human bone specimen, which included A-W GC particles (300-700 microns in diameter) implanted as a bone filler for about 75 weeks was harvested and investigated. Excellent A-W GC-bone bonding was observed, and the ultrastructure of the interface was similar to that in rats after long-term implantation. This finding demonstrated that A-W GC possibly worked in human bone in the same way as in rat bone, showing excellent bioactivity.

Adult↗

Repair of osteochondral defects with grafts of cultured chondrocytes. Comparison of allografts and isografts.

Isogeneic and allogeneic chondrocytes cultured in collagen gel were transplanted into osteochondral defects in knee joints of inbred strains of rats. At 12 weeks, all eight isografted defects had healed successfully, compared with only four of eight allografted defects (p < 0.05). At 26 and 52 weeks, all defects except one had healed successfully and there was no significant difference in the success rate between the isografted and allografted groups. Control defects that had been implanted with only collagen gel did not heal successfully.

Animals↗

Apatite formation on three kinds of bioactive material at an early stage in vivo: a comparative study by transmission electron microscopy.

Apatite formation on the surface of three kinds of bioactive material at an early stage after implantation in bone was studied using transmission electron microscopy (TEM). The materials were apatite- and wollastonite-containing glass-ceramic (A-W GC) as a surface-active glass-ceramic, dense sintered hydroxyapatite (HA) as a surface-active ceramic, and dense sintered beta-tricalcium phosphate (beta-TCP) as a resorbable ceramic. Particles of these materials, ranging from 100-300 microns in diameter, were implanted into rat tibiae, and specimens were prepared at 3, 7, 10, and 14 days after implantation. For A-W GC, dissolution of the glassy and probably wollastonite phase was observed in the surface region on and after the third day, and a collagen-free thin apatite layer on the surface of the material was evident on and after the seventh day. This apatite layer was observed before the mineralization of the surrounding bone matrix and was sometimes evident even where the material bordered on the bone marrow. On and after the tenth day, the surrounding bone matrix calcified and A-W GC-bone bonding through an apatite layer was completed. For HA, a mineralized collagen-free layer was observed on the surface of the ceramic on and after the tenth day. This layer was always present near calcifying bone and it was difficult to distinguish from immature bone. For beta-TCP, such a surface mineralized layer was rarely evident, even just before bone-ceramic contact, and finally the bone bonded to beta-TCP directly. Cell-mediated degradation of beta-TCP was frequently observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Differences in ceramic-bone interface between surface-active ceramics and resorbable ceramics: a study by scanning and transmission electron microscopy.

The interface between bioactive ceramics and bone was studied by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The materials were apatite-wollastonite-containing glass ceramic (A-W.GC) as a representative surface-active ceramic, and calcite and beta-tricalcium phosphate (beta-TCP) as resorbable ceramics. Particles of these materials, ranging between about 100 microns and 300 microns in diameter, were implanted into rat tibiae, and specimens were prepared for observation at 8 weeks after implantation. Both SEM and TEM demonstrated that A-W.GC was bonded to bone through a thin Ca-P-rich layer consisting of fine apatite crystals apparently different from those of bone in shape, size, and orientation. Collagen fibers of the bone reached the surface of this layer, and chemical bonding between A-W.GC and the bone was speculated. Calcite and beta-TCP, on the other hand, made direct contact with the bone, and no apatite layer was present at the interface. The surfaces of the implants became rough due to degradation, and bone grew into the finest surface irregularities. However, we were unable to demonstrate any continuity of crystals between the resorbable implants and bone by high-resolution TEM. Accordingly, the bonding strength was considered to be mainly attributable to mechanical interlocking.

Animals↗

A comparative study of ultrastructures of the interfaces between four kinds of surface-active ceramic and bone.

The interfaces between four kinds of surface-active ceramic and bone were studied by scanning electron microscopy (SEM) and transmission electron microscopy (TEM) using undecalcified specimens. The materials were Bioglass-type glass (Bioglass), Ceravital-type glass-ceramic (KGS), apatite- and wollastonite-containing glass-ceramic (A-W.GC) and hydroxyapatite (HA). Particles of these materials, ranging between about 100 and 300 microns in diameter, were implanted into rat tibiae, and specimens were prepared for observation at 8 weeks after implantation. All materials were observed to bond to bone through a collagen-free layer consisting of fine apatite crystals distinct from those in bone. The crystals of this apatite layer and those of bone were intermingled at their interface, suggesting chemical bonding. In Bioglass, which had only a glassy phase, several tens of microns of the material surface had changed to such an apatite layer. In KGS and A-W.GC, which had macrocrystals in the glassy phase, an intervening apatite layer about 0.5 micron thick was observed between the materials and bone. Furthermore, fine apatite crystals were also observed among the macrocrystals near the surface of the materials. In HA, which had no glassy phase, an intervening apatite layer was much less distinct and sometimes absent. These differences were considered to be attributable to the differences in chemical composition, crystallization, and solubility of the materials.

Animals↗