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[Experimental investigation on application of hydroxyapatite implant to alveolar ridge augmented by porous hydroxyapatite granules--histological observation on implant and implant covered with autogenous iliac bone].

UNLABELLED: The purpose of this study was to investigate the possibility of applying the hydroxyapatite implant and the hydroxyapatite implant covered with autogenous iliac bone to the augmented mandible. Three months after the extractions of the lower P3, P4, and M1 of mongrel dogs, alveolar ridge augmentations were performed widely with porous hydroxyapatite granules. After one month, the hydroxyapatite implants were inserted in the iliac bone of the same dog. Four months after the augmentation, the hydroxyapatite implant and the hydroxyapatite implant covered with autogenous iliac bone were implanted only in the augmented area with the granules of the mandible. Two months later, specimens were taken out and fixed by 10% formalin alcohol. They were embedded in polyester resin and undecalcified sections were prepared. The sections were stained with toluidine blue and observed under light microscope. RESULTS: 1) Bone ingrowth was seen in most parts of the intergranular spaces and in some spaces fibrous connective tissues were observed. 2) The hydroxyapatite implant was partly attached to the bone. But a large surface of the implant was connected to the fibrous tissues among the granules. 3) The hydroxyapatite implant covered with the iliac bone was combined with the bone of the intergranular spaces.

Alveolar Process

Hyaluronan interactions with hydroxyapatite do not alter in vitro hydroxyapatite crystal proliferation and growth.

The interaction of hyaluronan (Mr range 80-120 x 10(4)) with poorly crystalline hydroxyapatite, such as is found in calcified cartilage and bone, was studied to challenge the hypothesis that free hyaluronan found in proteoglycan aggregate preparations could affect in vitro mineralization. Using a Langmuir adsorption isotherm, based on uronic acid content, hyaluronan was found to bind to hydroxyapatite with an affinity K of 0.12 ml/microgram uronate and N = 6.8 micrograms uronate/m2 hydroxyapatite binding sites. This is contrasted with K = .047 ml/microgram uronate and N = 9.0 micrograms uronate/m2 for a bovine nasal proteoglycan monomer preparation. Although the proteoglycan monomer and aggregate preparations have been reported to inhibit hydroxyapatite growth at concentrations of 1 mg/ml, using solution concentrations of 0, 0.01, 0.1 and 1 mg/ml hyaluronan there were no detectable alterations in the rate of seeded hydroxyapatite growth and proliferation. These data indicate that although in vitro hyaluronan may bind with weak affinity to hydroxyapatite, this interaction does not affect mineral growth, and the presence of hyaluronan would not contribute to the increased inhibitory potential of cartilage proteoglycan aggregate relative to monomer preparations.

Adsorption

Interface between bone tissue and implants of solid hydroxyapatite or hydroxyapatite-coated titanium implants.

Loaded prestressed implants of dense hydroxyapatite and non-loaded hydroxyapatite-coated titanium implants were placed in edentulous regions of the lower jaw of dogs. After 6 month the jaw specimens were fixed and embedded in methyl-methacrylate. Thin non-decalcified ground sections were made for histology. Although the hydroxyapatite showed histological differences between the coated implants and the prestressed solid ones, both had an extensive apposition of normal lamellar bone on the whole surface of the bone-buried part of the implant. The bone contact was very intimate and without any visible intermediate tissue layer. The tissue response observed forms a good biological base for the clinical application of hydroxyapatite-coated titanium implants.

Animals

[Study on hybrid materials compounded with porous hydroxyapatite and culture cells. 1. Attachment and the invasion of cultured cells to porous hydroxyapatite].

The purpose of this study was to investigate the effects of hybrid material compounded with periodontal ligament cells on human teeth and porous hydroxyapatite. As the first step, we tried to produce hybrid materials compounded with periodontal ligament cells and porous hydroxyapatite, and to observe the attachment and invasion of periodontal ligament cells porous hydroxyapatite. Human periodontal ligament cells which had been scraped off the root with a disposable blade were cultured at 37 degrees C in alpha-MEM plus 10% fetal calf serum. The culture cells were attached to the porous hydroxyapatite in vitro. After 3 and 14 days of incubation, the specimens were fixed in gluteraldehyde and examined using SEM. Other specimens were embedded in paraffin and successive specimens were prepared for to remodelling of the attachment and invasion. Many cells attached to the P-HAP granules, and some cells invaded the P-HAP granules. The results suggest that when these hybrid materials are implanted in bone defects, the cells remain in the defect for a certain period, and exhibit the functions and characteristics of itself.

Bone Regeneration

Fibroblastic growth and attachment on hydroxyapatite-coated titanium surfaces following the use of various detoxification modalities. Part II: Contaminated hydroxyapatite.

This study evaluated the ability of various chemotherapeutic and mechanical modalities to detoxify endotoxin-contaminated hydroxyapatite-coated dental implant surfaces as determined by the early attachment and growth of human gingival fibroblasts. Hydroxyapatite-coated test strips were contaminated with purified outer membranes of Escherichia coli and treated with citric acid, hydrogen peroxide, stannous fluoride, chlorhexidine gluconate, tetracycline HCl, polymyxin B, a plastic sonic scaler tip, or left untreated (contaminated and sterile controls). Human gingival fibroblasts were then seeded onto the test strips and incubated for 48 hours. The citric acid-treated strips showed greater cell growth than the other treatments. The plastic sonic scaler tip and the polymyxin B-treated samples exhibited greater cell coverage than the sterile control specimens. The use of citric acid and/or a modified plastic sonic scaler tip may be a valuable adjunct when surgical repair of an ailing hydroxyapatite-coated dental implant is contemplated.

Cell Adhesion

Interaction of hydroxyapatite and protein-coated hydroxyapatite with Streptococcus mutans and Streptococcus sanguis.

The present study showed that S. mutans and S. sanguis behaved like negatively-charged particles in their interaction with hydroxyapatite in vitro. Phosphate in the system inhibited bacterial uptake by apatite, whereas calcium increased the uptake. A layer of acidic protein inhibited the uptake of bacteria by hydroxyapatite. The opposite was true when a basic protein was first adsorbed to the apatite. A saliva film on the apatite decreased the uptake of bacteria, supporting the view that acidic proteins are selectively adsorbed by hydroxyapatite from saliva. The results indicate clearly that electrostatic forces may be involved in bacterial interaction with tooth surface.

Adsorption

Selective adsorption of porcine-amelogenins onto hydroxyapatite and their inhibitory activity on hydroxyapatite growth in supersaturated solutions.

The selective adsorption of amelogenins onto synthetic hydroxyapatite (HA) and their inhibitory activity on the seeded HA crystal growth were investigated using enamel proteins obtained from the outer layer of immature porcine-enamel (soft, cheeselike in consistency) of developing permanent incisors. Special interests were paid to the effect of a postsecretory degradation of the original amelogenin(s) on their adsorption and inhibitory activity. In the adsorption studies, it was apparent that the originally secreted amelogenin (25 K), proline, and histidine-rich protein (2a), as well as the higher molecular weight components (60-90 K), showed a strong adsorption affinity onto the HA. This adsorption of protein 2a was related to its inhibition of the crystal growth of seeded HA in a dilute supersaturated solution. On the other hand, the partially degraded product (20 K) of amelogenins, protein 2b, lost the high adsorption affinity onto the HA, and consequently showed no significant inhibitory activity. The observed selective adsorption of protein 2a onto HA was apparent at pH 6.0 and pH 7.4 even in the presence of dissociative solvents, such as 3 M urea or 2 and 4 M guanidine-HCl; however, this selective behavior was sensitive to changes in pH, and was not displayed at pH values of 7.8 or 10.8. The results suggest that the originally secreted amelogenin 2a may play an active role in amelogenesis, and that enamel mineralization could be regulated by the secretion of amelogenins and their inactivation through partial enzymic degradation, prior to their complete removal.

Adsorption

Adhesion of carboxylate cements to hydroxyapatite. I. The effect of the structure of aliphatic carboxylates on their uptake by hydroxyapatite.

An investigation was made into the effect of the structure and functionality of aliphatic carboxylates on their sorption onto hydroxyapatite with a view to ascertaining factors affecting the adhesion of polyacrylate cements to tooth materials. In general, the amount of carboxylate sorbed was found to increase with the number of groups contained in the molecule. Thus, polyacrylate was found to be much more strongly sorbed than low molecular weight species. Sorption of the low molecular weight species appeared to be related to stereochemical factors rather than to the stability constants of their calcium chelates.

Adsorption

Fibroblastic growth and attachment on hydroxyapatite-coated titanium surfaces following the use of various detoxification modalities. Part I: Noncontaminated hydroxyapatite.

Hydroxyapatite-coated titanium alloy test strips were treated with chlorhexidine gluconate, stannous fluoride, citric acid, tetracycline HCl, polymyxin B, hydrogen peroxide, and a plastic Cavitron tip: untreated sterile strips served as controls. The strips were incubated with cultured human gingival and periodontal ligament fibroblasts. Image analysis of three photomicrographs of each test strip (original magnification x350) indicated that the tetracycline HCl treatment resulted in significantly greater cellular surface area coverage compared with the other treatments. Citric acid and the plastic Cavitron tip also stimulated cell attachment, although the results from the Cavitron tip were not significantly different from citric acid or the other treatment groups. The remainder of the modalities and the untreated cellular controls experienced similar cellular coverage.

Cell Adhesion

Reaction of bone to HA, carbonate-HA, hydroxyapatite + calcium orthophosphate and to hydroxyapatite + calcium ortho- and pyrophosphate.

Slip cast conical implants of HA, carbonate-HA, calcium orthophosphate/hydroxyapatite and hydroxyapatite/calcium pyrophosphate/calcium orthophosphate with weight ratios 75/25 and 50/30/20 were implanted in rabbit tibia. The bone-implant interfaces were evaluated histologically, by means of SEM/EDX analysis and by push-out test. Thirty-six implanted samples were investigated after 2, 8, and 24 weeks. The triphasic calcium phosphate showed a moderate disintegration. This material showed 2 weeks after implantation a bonding between new bone and implant could be seen in parts of the operation site by SEM. Eight weeks after implantation an intimate relationship between the bone tissue and all specimens of each material was found. After 24 weeks the Ca/P ratio in the bone had reached the Ca/P ratio of mature bone determined by SEM/EDX analysis.

Animals

Adherence of Streptococcus sanguis to hydroxyapatite coated with lysozyme and lysozyme-supplemented saliva.

The adherence of [3H]thymidine-labeled Streptococcus sanguis strains to bare hydroxyapatite and to hydroxyapatite coated with a range of concentrations of lysozyme, poly-L-lysine, poly-L-glutamic acid, whole saliva supernatant, and combinations of some of the above was studied. Adherence of several strains of S. sanguis to bare hydroxyapatite and saliva-coated hydroxyapatite was compared. Saliva present as a pellicle on the hydroxyapatite inhibited adherence of some strains (903, M-5, 73X11) and stimulated that of others (S35, B-4, 66X49). Strains 903 and S35 were chosen for further study. Adherence of both strains was stimulated up to fivefold by the presence of adsorbed lysozyme or poly-L-lysine on the hydroxyapatite, whereas poly-L-glutamic acid inhibited adherence (80 to 95%). Adherence of strain S35 to hydroxyapatite coated with combinations of saliva and (i) lysozyme, (ii) poly-L-lysine, or (iii) poly-L-glutamic acid was unaffected compared with adherence to hydroxyapatite coated with saliva alone. In contrast, adherence of strain 903 to hydroxyapatite coated with combinations of saliva and either lysozyme or poly-L-lysine was inhibited up to ca. 90% compared with hydroxyapatite coated with saliva alone. Strain 903 was also unaffected by combinations of poly-L-glutamic acid and saliva on the hydroxyapatite. Adherent cells of both strains were completely (greater than 90%) eluted with high-ionic-strength buffer from either bare hydroxyapatite or hydroxyapatite coated with lysozyme alone. Adherent cells of strain S35 were only poorly eluted (25%) from hydroxyapatite coated with either saliva alone or saliva and lysozyme. Strain 903 elution from hydroxyapatite coated with either saliva alone or saliva and lysozyme was essentially complete. These observations were taken to indicate that the two test strains adhered to saliva-coated hydroxyapatite by different mechanisms. Protein-coated hydroxyapatite was shown not to be saturated under the conditions described here. Examination by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the variously supplemented salivary pellicles formed on the hydroxyapatite demonstrated that major changes in salivary protein composition did not occur when lysozyme, poly-L-lysine, or poly-L-glutamic acid was used to supplement saliva. Lysozyme-dependent aggregation of strain 903 was shown not to occur under the conditions of our experiments. We suggest that the basis for stimulation of adherence to hydroxyapatite coated only with lysozyme is an increase in the cationic surface area available for electrostatic adherence of the microorganisms.(ABSTRACT TRUNCATED AT 400 WORDS)

Adhesiveness

The mechanical properties and solubility of strontium-substituted hydroxyapatite.

The mechanical properties and solubility of sintered Sr-substituted hydroxyapatites were examined in order to investigate the influences of Sr incorporation into hydroxyapatite in bones and teeth on them. Hydroxyapatite (HAp), Sr-substituted hydroxyapatite (Ca.SrAp), and strontium-hydroxyapatite (SrAp) were synthesized and sintered. The bending strength and Young's modulus were measured. Also, solubility in isotonic NaCl solution was examined. The Young's modulus of Sr-substituted hydroxyapatite (Sr wt% = 1.0) and hydroxyapatite surface treated with Sr ion were greater than that of hydroxyapatite. The bending strengths of Sr-substituted hydroxyapatites (Sr/Ca = 4/6 and less) were reduced slightly compared to that of hydroxyapatite. The bending strength of hydroxyapatite surface treated with Sr ion was increased compared to that of hydroxyapatite. The solubility in isotonic NaCl solution at 37 degrees C was increased with an increase in Sr content. The effects of Sr on the mechanical properties and solubility of bones and/or teeth are discussed.

Biomechanical Phenomena