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Bone healing following the use of hydroxyapatite or ionomeric bone substitutes alone or combined with a guided bone regeneration technique: an animal study.

The healing of standardized bone defects grafted with either particulate ionomeric or hydroxyapatite bone substitutes was compared in the mandibular ramus of 30 Sprague-Dawley rats. The possible additional response achieved when combining these materials with a guided bone regeneration (GBR) technique was also evaluated. Three groups of 10 animals received either no implant material or ionomeric or hydroxyapatite bone substitute in defects in the right ramus. The left mandibular defects received the same treatment, except that the operation site was covered by a membrane (GBR technique). Half of the animals were sacrificed at 4 and 10 weeks following surgery, and the inflammatory response at the implant site and the amount of new bone formed in the defects were determined histomorphometrically. Defects implanted with ionomeric bone substitute exhibited more bone formation (4 weeks = 3.19 +/- 0.35 mm2, 10 weeks = 5.35 +/- 0.26 mm2) than both defects that received no treatment (4 weeks = 0.88 +/- 0.35 mm2, 10 weeks = 2.1 +/- 0.49 mm2), membrane alone (4 weeks = 1.21 +/- 0.05 mm2) or hydroxyapatite bone substitute (4 weeks = 1.41 +/- 0.46 mm2, 10 weeks = 3.34 +/- 0.41 mm2) at 4 weeks (P < or = .01) and at 10 weeks (P < or = .05). The use of a GBR technique did not increase the amount of bone formed, compared to the use of bone substitutes alone. Hydroxyapatite and ionomeric bone substitutes used alone were more effective in inducing repair of the defects than was GBR membrane alone. The use of hydroxyapatite was associated with a greater inflammatory reaction (P < or = .01) than was ionomer in this model.

Analysis of Variance↗

[A dynamic study of the intensity of mitogenesis in an osteoblast culture in the presence of ultrasonically dispersed hydroxyapatite].

Ceramic hydroxyapatite is frequently used in clinical practice nowadays; it is applied in surgical interventions on facial skull bones. This biological material is used as bone substitute and bone connector, but many authors claim that it is devoid of osteoinductor properties. Research is in progress on the effects of a "cold", that is, biochemically active form of ceramic hydroxyapatite on proliferative activity of a number of cell populations. No mitogenesis stimulation was observed during incubation of hydroxyapatite with osteoblast culture; on the contrary, a reduction of proliferative processes intensity was observed. Remembering that hydroxyapatite is a surfactant we repeated this experiment under similar conditions but used a hydroxyapatite molbfication with specific activity increased by two orders in comparison with its close analogs. Mitogenesis intensity was assessed by radiometric method from 3H-thymidine incorporation. The data indicated stimulation of proliferative processes in osteoblast culture.

Animals↗

Hydroxyapatite cement. I. Basic chemistry and histologic properties.

Hydroxyapatite cement is a unique calcium phosphate preparation that can be shaped intraoperatively and sets in vivo to an implant composed of microporous hydroxyapatite. The histologic response to this cement was evaluated by implanting disks made of this material within the heads of nine cats. Three sets of 12 hydroxyapatite cement disks were produced containing 0%, 10%, and 20% macropores by volume, respectively. The disks were implanted subcutaneously, intramuscularly, above the periosteum of the skull, and directly onto the surface of the calvarium. Each macropore percentage was represented in each tissue plane, and animals were killed up to 9 months postoperatively. There were no toxic reactions, implants extruded, or wound infections. Histologic examination of the implant-soft-tissue interfaces revealed a transient inflammatory response without foreign body reaction. The disks were resorbed over time in direct proportion to their macropore content (surface areas) in all groups except for those disks placed directly onto the surface of the calvarium below the periosteum. In this group, numerous foci of bone formed at the skull-implant interface, with variable replacement of the deep surface of these implants by bone. Implant replacement by bone is postulated to occur through a combination of implant resorption coupled with osteoconduction. Based on these properties, hydroxyapatite cement may prove useful when applied to the reconstruction of non-stress-bearing skeletal tissue.

Animals↗

Slow-releasing potential of vancomycin-loaded porous hydroxyapatite blocks implanted into MRSA osteomyelitis.

Although antibiotic-loaded hydroxyapatite blocks have been used for the treatment of chronic osteomyelitis, their long-term potential for releasing antibiotic into human bones is not well known. Five patients with chronic osteomyelitis due to methicillin-resistant Staphylococcus aureus (MRSA) infection were effectively treated with local implantation of vancomycin-loaded hydroxyapatite blocks. Blocks were removed during the following reconstructive surgeries when the releasing capability of the blocks, and the bacteriocidal activity of the remaining vancomycin in these blocks could be evaluated. Vancomycin was rapidly released within 1 month after implantation, and by 3 months 90% of vancomycin had leaked from the blocks. At 18 months vancomycin still remained in a bacteriocidal form in the hydroxyapatite blocks, though the blocks had no releasing potential or the eluted vancomycin had been changed to a different form. Vancomycin-loaded porous hydroxyapatite blocks would be useful for the treatment of chronic osteomyelitis or implant-associated osteomyelitis due to MRSA.

Adult↗

The biological performance of calcium phosphate ceramics in an infected implantation site: I. Biological performance of hydroxyapatite during Staphylococcus aureus infection.

In the present study the biological performance of macroporous and dense hydroxyapatite after implantation in the rat middle ear was evaluated during an induced Staphylococcus aureus middle ear infection. The course of the infection was similar to that in the absence of an implant. Hydroxyapatite was frequently integrated with fibrous ingrowths in the middle ear lumen, originating solely from the infection. Good epithelial covering of the implant with all types of epithelial cells of importance for middle ear defence, was found. Increase of the exudate in the pores due to the infection was relatively small, and most of the exudate was restricted to pores on the implant surface. The bony tissue in the pores was not influenced significantly by the induced infection. Degradation of hydroxyapatite was consistent with earlier results obtained in the non-infected middle ear. The results obtained so far suggest that hydroxyapatite is highly suitable for middle ear implantation.

Animals↗

The biocompatibility of hydroxyapatite ceramic: a study of retrieved human middle ear implants.

The biocompatibility of 11 hydroxyapatite auditory canal-wall prostheses and 4 hydroxyapatite incus prostheses implanted for 4 to 40 months was evaluated by light microscopy, scanning electron microscopy, transmission electron microscopy, and Röntgen microanalysis. These 15 prostheses representing 4% of 375 prostheses, has been removed because of unresolved chronic middle ear infection, residual cholesteatoma, or poor fit. The findings confirmed earlier reports on the biocompatibility of hydroxyapatite in vitro, in animals, and in man. An electron-dense layer was found at the interface with bone and fibrous tissue, and a firm bond between the ceramic and bone at the hydroxyapatite ceramic/bone interface developed. Macropores became filled with bone and fibrous tissue, and the tissue in the individual pores was interconnected. Furthermore the incus prostheses were covered with an epithelium similar to that found in the human middle ear. Findings diverging from those made in other studies were the relatively large amount of exudate in the pores, an apparent increase of degradation during infection, and the accumulation of trace elements in one of the canal-wall prostheses. In all likelihood these three phenomena may be attributed to the unfavorable conditions to which these prostheses were exposed during implantation.

Biocompatible Materials↗

In vitro bioactivity of silicon-substituted hydroxyapatites.

Silicon-containing hydroxyapatites were synthesized by the controlled crystallization method. Chemical analysis, N(2) adsorption, Hg porosimetry, X-ray diffraction, scanning electron microscopy-energy-dispersive X-ray spectroscopy, and X-ray photoelectron spectroscopy (XPS) were used to characterize the hydroxyapatite and to monitor the development of a calcium phosphate layer onto the surface of the substrate immersed in a simulated body fluid, that is, in vitro bioactivity tests. The influence of the silicon content and the nature of the starting calcium and phosphorus sources on the in vitro bioactivity of the resulting materials were studied. A sample of silicocarnotite, whose structure is related to that of hydroxyapatite and contains isolated SiO(4) (4-) anions that isomorphically substitute some PO(4) (3-) anions, was prepared and used as reference material for XPS studies. An increase of the unit cell parameters with the Si content was observed, which indicated that SiO(4) (4-) units are present in lattice positions, replacing some PO(4) (3-) groups. By using XPS it was possible to assess the presence of monomeric SiO(4) (4-) units in the surface of apatite samples containing 0.8 wt % of silicon, regardless the nature of the starting raw materials, either Ca(NO(3))(2)/(NH(4))(2)HPO(4)/Si(OCOCH(3))(4) or Ca(OH)(2)/H(3)PO(4)/Si(OCOCH(3))(4). However, an increase of the silicon content up to 1.6 wt % leads to the polymerization of the silicate species at the surface. This technique shows silicon enrichment at the surface of the three samples. The in vitro bioactivity assays showed that the formation of an apatite-like layer onto the surface of silicon-containing substrates is strongly enhanced as compared with pure silicon-free hydroxyapatite. The samples containing monomeric silicate species showed higher in vitro bioactivity than that of silicon-rich sample containing polymeric silicate species. The use of calcium and phosphate salts as precursors lead to materials with higher bioactivity.

Calcium↗

Bovine serum albumin conformational changes upon adsorption on titania and on hydroxyapatite and their relation with biomineralization.

The biocompatibility of implant materials used for substitution of bone tissue depends on its ability to induce the deposition of a hydroxyapatite layer when in contact with body fluids. In previous work, some of the authors found that bovine serum albumin (BSA) promotes calcium phosphate deposition if preadsorbed on hydroxyapatite and retards precipitation if preadsorbed on titania. In the present study, we investigated the adsorption of BSA upon particles of titania and hydroxyapatite in order to understand the different role played by the protein on the mineralization of both biomaterials. The adsorption isotherms were determined and the structural changes induced by adsorption at different surface coverages were investigated by circular dichroism spectroscopy and differential scanning microcalorimetry. At low surface coverages, the adsorbed BSA molecules lost part of their alpha-helix content. However, at high surface coverages, corresponding to the plateau values of the adsorption isotherms, the BSA molecules did not undergo structural rearrangements upon adsorption. In the latter circumstances, the availability of BSA calcium binding sites, which should be responsible for inducing mineralization, depends on the electrostatic interactions between BSA and the sorbent surface. A possible explanation for the different mineralization behavior of hydroxyapatite and titania is advanced.

Adsorption↗

Electrochemical processes of nucleation and growth of hydroxyapatite on titanium supported by real-time electrochemical atomic force microscopy.

Recently, interest in electrochemical formation of hydroxyapatite has evolved. In this work, highly crystalline hydroxyapatite is electrodeposited on pure titanium and Ti-6Al-4V alloy. In situ and ex situ imaging, coupled with potentiostatic and potentiodynamic measurements, is conducted by means of electrochemical atomic force microscopy. This allows for a study of the nucleation and growth of hydroxyapatite as well as of its near-atomic structure. Electrodeposition of hydroxyapatite is shown to result from precipitation in solution, following two stages: (1) instantaneous nucleation, two-dimensional growth; (2) progressive nucleation, three-dimensional growth. Although some nucleation occurs already at -842 mV, potentials that are more negative than -1.26 V versus SCE are required for enhanced growth. Mass transport is found to have only secondary effect on the deposition process. The conclusions of this work have implications in optimization of coatings on implants as well as in enhancement of the understanding of bone mineralization in vivo.

Biomimetic Materials↗

Preparation of needle-like hydroxyapatite by homogeneous precipitation under hydrothermal conditions.

Preparation of hydroxyapatite was carried out by a homogeneous precipitation technique using hydrothermal reactions of Ca(EDTA)2- and PO4(3-) at 130-250 degrees C and pH 4-10. Hydroxyapatite and monetite were formed above pH 6 and below pH 4, respectively. Needle-like hydroxyapatite was formed at high temperatures above 130 degrees C. The length of the needle-like hydroxyapatite crystal increased with increasing working solution pH and temperature.

Calcium↗

Sintered hydroxyapatite for a percutaneous device and its clinical application.

The sintered hydroxyapatite was designed to be utilized as a percutaneous device. The device was percutaneously fixed through the back skin of mongrel dogs. The tissue reaction around the implants was examined histologically. At 1 month the sintered hydroxyapatite was closely contacted with the skin tissue and downgrowth of epidermis was not observed. The long-term implantation ranging from 3 to 17 months showed that the depth of epidermis downgrowth was limited to only 1 mm. The amino acid composition of fibrous capsule formed around the hydroxyapatite showed close resemblance to vertebrate periosteum. It was confirmed that the sintered hydroxyapatite had good compatibility and long-term stability with skin tissue.

Amino Acids↗

Hydroxyapatite ceramic as a bone substitute.

We have used hydroxyapatite ceramic as a bone substitute for grafting in extensive bone replacement. Hydroxyapatite is composed of calcium phosphate and is incorporated into bone as a physiological mineral. It is not antigenic, carcinogenic or osteogenic. The material used in this study differs from other hydroxyapatite ceramics in that it is an interconnected porous system. Between 1981 and 1986 we used this material on forty five occasions in 44 patients. It was combined with autologous cancellous bone graft in 38 cases and used alone in a further seven. It was usually employed to fill defects after removal of bone cysts and for long fusions in patients with scoliosis. It was also used to fill bone defects after trauma, for non-union in the lumbosacral area, for anterior vertebral fusions, in limb sparing operations for malignant bone tumours, and in exchange operations after failed joint endoprostheses. We have not seen incompatibility or rejection of the implanted material. The rate of postoperative infection was higher than usual due to the selection of patients, but did not appear to be connected with the use of hydroxyapatite ceramic. We were able to review 36 patients up to sixty months after operation. The results are encouraging with no difference in progress compared with patients in whom simple autologous bone grafts had been used.

Adolescent↗

MRI of orbital hydroxyapatite implants.

Our aim was to use MRI for the postsurgical assessment of a new form of integrated orbital implant composed of a porous calcium phosphate hydroxyapatite substrate. We studied ten patients 24-74 years of age who underwent enucleation and implantation of a hydroxyapatite ball; 5-13 months after surgery, each patient was examined by spinecho MRI, with fat suppression and gadolinium enhancement. Fibrovascular ingrowth was demonstrated in all ten patients as areas of enhancement at the periphery of the hydroxyapatite sphere that extended to the center to a variable degree. The radiologist should aware of the MRI appearances of the coralline hydroxyapatite orbital implant since it is now widely used following enucleation. MRI is a useful means to determine successful incorporation of the substrate into the orbital tissues. The normal pattern of contrast enhancement should not be mistaken for recurrent tumor or infection.

Adult↗

Template-directed synthesis and selective adsorption of oligoadenylates on hydroxyapatite.

Polyuridylic acid is adsorbed completely from aqueous solution by hydroxyapatite under conditions that permit template-directed synthesis of oligoadenylates in free solution. The yield of oligoadenylates is enhanced to almost the same extent by poly(U) in the presence or the absence of hydroxyapatite. Under very similar conditions small quantities of hydroxyapatite adsorb higher molecular-weight oligoadenylates selectively from a mixture of oligomers. On the basis of these results we propose a mechanism for prebiotic oligonucleotide formation in which selective adsorption on hydroxyapatite or some other immobilized anion-exchanging material plays a major role. Monomers are released from the surface for reactivation, while oligomers are retained in a protected environment by adsorption to the apatite surface.

Adenine Nucleotides↗

The interaction of collagenase with hydroxyapatite and related materials and enzymatic properties of the adsorbed enzyme.

The commercial collagenase from Clostridium histolyticum was adsorbed on hydroxyapatite, on bovine femur shaft and on enamel and dentin powders. The substrate specificity of the adsorbed enzyme tested, with chromophore substrate, azocoll and native collagen, differed from that obtained with the soluble enzyme. The adsorption and the substrate specificity was also dependent on the adsorbent used. A pretreatment of hydroxyapatite with chondroitin sulphate, hyaluronate and DNA lowered the adsorption of collagenase. Phosphate ion caused desorption of the enzyme from hydroxyapatite. Sodium fluoride caused partial desorption of the enzyme from hydroxyapatite and enamel and dentin powders. Colangenase adsorbed on root surfaces of teeth liberated hydroxyproline containing material.

Adsorption↗

Effects of pyrophosphate and diphosphonates on the dissolution of hydroxyapatites using a flow system.

Pyrophosphate and diphosphonate ions have been said to diminish the dissolution of hydroxyapatite crystals, because they lower the equilibrium concentrations of calcium and phosphate ions in the bulk solution around hydroxyapatite crystals in a closed system. However, in a closed system these effects are not necessarily due to an effect on dissolution alone. In this paper we have used a continuous flow system to study the effects of pyrophosphate and two diphosphonates, ethane-1-hydroxy-1,1,-diphosphonate and dichloromethane diphosphonate, on the dissolution of hydroxyapatite. All three compounds decreased markedly the rate of dissolution of hydroxyapatite as well as the exchangeable pools of calcium and phosphate ions around the crystals.

Calcium↗

The effect of locally implanted complexes of adriamycin-hydroxyapatite complexes on tumor--study of a new alternative treatment for hepatic cancer.

A new targeting anticancer system was prepared by using hydroxyapatite particles (2 mm in diameter) as carrier material and adriamycin as anticancer agent. The adsorption and release properties of the complexes were assayed by fluorometry in vivo and in vitro and the curative effect on W-256 sarcoma of rat was observed. The results showed that one particle of hydroxyapatite could adsorb approximately 0.08 mg adriamycin and they can maintain a steady and slow release of adriamycin from hydroxyapatite for one month. When hydroxyapatite-adriamycin complexes were implanted into the liver of rat, liver adriamycin concentration at the implanted region was obviously higher than that achieved by injection of adriamycin solution. The locally implanted complexes obviously inhibited the growth of subcutaneous implanted tumor of rat, and increased the survival rate of rat with implanted liver tumor.

Animals↗

The interaction of proteins with hydroxyapatite. II. Role of acidic and basic groups.

The elution behavior from hydroxyapatite columns of the modification products of seven basic and three acidic proteins has been investigated. Three classes of NH2 derivatives were prepared. These consisted of (1) replacement by a guanidyl group with no change in charge; (2) blocking with loss of charge; and (3) replacement of positive charges by negative ones. Two types of COOH derivatives were prepared: (1) blocking with loss of charge; and (2) replacement of COOH by SO3H with no change in charge. The elution behavior of the derivatives in PO4, F-, Cl-, ClO-4, and Ca2+ ion eluants showed that (1) the elution patterns are determined by the isoelectric points of the proteins, there being no symmetry between the binding or elution behavior of acidic and basic proteins; (2) the binding of basic proteins requires the presence of a high density of positively charged groups; (3) the binding of all proteins to hydroxyapatite equilibrated with phosphate buffer is enhanced by a decrease in the number of their negative charges; and (4) calcium ions affect the binding of proteins to hydroxyapatite at the level of carboxyls, since clusters of carboxyls strengthen both the interaction with Ca2+ and the binding to hydroxyapatite.

Chemical Phenomena↗