COMPOSITION FOR RESTORING DEFECTS IN CALCIFIED TISSUES

Abstract: A composition for restoring defects in calcified tissues comprises an amorphous bioactive particulate consisting essentially of calcium, silicon and oxygen. Said agent may be selected from the group consisting of : a) a reaction product of an organic silicate source and a source of calcium; b) a calcium containing hydrolysis product of tetraethylorthosilicate; c) a calcium containing silica sol-gel; d) a binary calcium oxide and silicate precipitated material; e) a synthetic analog of a naturally occurring wollastonite-like calcium silicate; and f) a precipitated reaction product of soluble calcium source and a silicate solution. A process for producing the amorphous bioactive particulate consists essentially of adding a sufficient amount of a solution of a soluble calcium source to a solution of a silicate or silicate precursor to cause a precipitation of calcium-containing silicate particulate.

Description: COMPOSITION FOR RESTORING DEFECTS IN CALCIFIED TISSUES FIELD OF THE INVENTION The present invention relates generally to compositions of amorphous bioactive particuiates consisting essentially of calcium, silicon and oxygen and methods for their use in restoring defects in calcified tissues. BACKGROUND OF THE INVENTION It is known that products such as bioactive particuiates can be used in restoring or repairing defects in calcified tissues such as periodontal pockets, extraction sites, treatments of dentin hypersensitivity, bone tissue formation and the like. The biocompatability of bioactive particuiates in restoring or repairing defects in calcified tissues such as bone tissue formation, as evidenced by tight apposition of hard tissue to implanted glassy particles, was first demonstrated in 1971. Specifically, the use of bioactive glass was originally reported by Hench et al. in J. Biomed. Mater. Res. Symp., 2: 117-141 (1971). Bone grows by apposition onto bioactive glass placed adjacent to bony surfaces and new bone may be conducted subsequently, over short distances. Granules of bioactive glass have been described in subsequent bone tissue formation applications. U.S. Pat. No. 4,239,113 describes a composition for the preparation of bone cement. U.S. Pat. No. 5,658,332 discloses a method to form osseous tissue in defects in sites in the appendicular skeleton or sites exhibiting reduced metabolic state using bioactive glass granules containing 40 to 58% Si0 2 , 10 to 30% NaO, 10 to 30% CaO, and 0 to 10% P 2 O 5 , and in the size range of from 200 to 300 micrometers. In the area of repairing or restoring defects in dental tissues, bioactive paniculate has been used for periodontal osseous defect repair (U.S. Pat. No. 4,851,046) utilizing a size range of 90-710 μm and a compositional range of 40-55 wt. % SiO 2) 10-30 wt. % CaO, 10-35 wt. % Na 2 0, 2-8 wt. % CaF 2 , and 0-10 wt. % B 2 0 3 . U.S. Pat. No. 4,851,046 describes compositions containing particles of bioactive glass of various size ranges for use in periodontal applications. U.S. Pat. No. 5,204,106 discloses that particles of bioactive glass within the narrow size range of 280 to 425 micrometers elicit a distinctively altered biologic response from that described previously. Bioactive particuiates have also been used in the treatment of dentin hypersensitivity. Dentin is a hard but resilient calcareous tissue that mechanically supports the dental enamel. The dentin encloses and protects the pulp. The dentin also has a large number of continuous dentinal tubules which connect the pulp cavity with the dentinoenamel/cementum junction. These fine dentinal tubules radiate out from the pulp. The problem of tooth sensitivity arises when the enamel and cementum are worn away from the tooth by either improper oral hygiene practices or as a result of periodontal disease and/or its σeatment. The dentinal tubules are fluid- filled exposing the central core of the tooth to the external environment. Hence, when subjected to air current, tactile or thermal stimulation, the pulpal nerves are irritated, leading to the perception of pain. Under a predominant theory called the hydrodynamic theory to explain the reason for hypersensitive dentin, the stimuli conveyed through dentinal tubules irritates nerves distributed in the dental pulp causing pain in these areas. Suppressing sensitive dentin and ameliorating or alleviating the pain might be achieved by sealing the dentinal tubules, thus blocking the conductor of the sensation by physical or chemical means. In earlier applications to provide for partial occlusion of dentinal tubules, U.S. Patent No. 5,735,942 ('"942 patent") discloses a bioactive melt-derived glass composition for the treatment of tooth hypersensitivity. The '942 patent discloses the following composition by weight percentage: 40-60 Si0 2 , 10-30 CaO, 10-35 Na 2 0, 2-8 P 2 0 5 , 0-25 CaF 2 and 0-10 B 2 0 3 , with particle size of less than about 10 μm with some particles are about 2 μm or less and some larger than 2 μm. The '942 patent notes that 60% SiO 2 is the maximum silica limit for bioactive melt- derived glass. In a melt-derived process, there are three reactions which lead to the development of a porous hydrated silica-gel layer: ion exchange, silica dissolution, and condensation of silanols to form siloxane-bonded hydrated silica chains or rings. As the Si0 2 content increases in melt-derived glasses, the rates of these reactions decrease, reducing the availability of Ca + ions in solution and the ability to develop the silica-gel layer on the surface. The result is the reduction and eventual elimination of the bioactivity of the melt-derived glasses as the Si0 2 content approaches 60%. The process imposes narrow effective compositional zones which hinder the formulator's ability to modify and tailor the material for a specific application. Bioactive glasses prepared by the melt-derived process and as taught in prior art ('942 patent) are processed in a platinum crucible at high temperatures, typically about 1350°C or so. Therefore, melt-derived glasses are costly and difficult to transfer to production facilities. The high- temperature processing plus the multiple handling steps render bioactive glass prepared by the melt-derived process rather expensive, effectively limiting the practicality of using bioactive glass for oral care products. Besides the high production cost, there are other disadvantages inherent with the prior art. First, it is difficult to maintain the very high purity required for optimal bioactivity due to the melt- derived process itself and the low silica and high alkali content of traditional bioactive glass compositions. Secondly, the prior art requires processing steps which involve grinding, polishing, fritting, sieving, etc., all of which expose the bioactive powder to potential contaminants that may negatively effect bioactivity. Third, there is a compositional limitation imposed on bioactive glasses and glass-ceramics made by conventional high-temperature processes due to the extremely high equilibrium liquidus temperature of Si0 2 of 1713°C, and the extremely high viscosity of silicate melts with high Si0 2 content. In another reference, US Patent No. 5,874,101 (the '"101 patent") proposes a bioactive gel prepared by an improved process, a sol-gel process which includes a drying step for the treadnent of tooth hypersensitivity. As with existing bioactive glasses and glass/ceramics and as disclosed in the the '942 patent, the bioactive glasses in the '101 patent are confined to a specific compositional zone in the Na 2 0-CaO-P 2 θ5-Siθ2 system defined mainly in silica content (Ogin, M., Ohuchi, F and Hench, L.L., Compositional dependence of the formation of calcium phosphate films on bioglass, J. Biomed. Mater. Res. 1980, 14, 55-64 and Ohtsuki, C, Kokubo, T., Takatsuka, K. and Yamamuro, T., Compositional dependence of bioactivity of glasses in the system CaO-Siθ 2 -P 2 0 3 : its in vitro evaluation, J. Ceram. Soc. Japn. 1991, 99, 1-6). The '101 patent discloses a composition in weight percent of 40-90 Si0 2 , 4-45 CaO, 0-10 Na 2 0, 2-16 P 2 0 5 , 0-25 CaF 2 , 0-4 B 2 0 3 , 0-8 K 2 0 and 0-5 MgO. None of the foregoing prior art references provide bioactive particuiates which are simple and easy to prepare, and which promote the initiation of caicium-contaimng mineral to restore and/or repair calcified mineral tissues. SUMMARY OF THE INVENTION This invention relates to an inorganic or organic / inorganic composite composition which consists essentially of CaO and Siθ 2 for restoring defects in calcified tissues. The organic/inorganic composite material is preferably an amorphous binary oxide material of the system CaO-Siθ 2 having been prepared by the addition of a soluble calcium source to a suitable silicon-donating precursor, such as TEOS or sodium silicate. In a preferred embodiment, the invention relates to an inorganic or organic / inorgamc composite composition which consists essentially of CaO and SiO 2 , which, when lodged in the tubules initiates the formation of calcium-containing mineral inside the dentinal tubules for the alleviation of the pain associated with sensitive teeth. Preferably the composite composition should have a particle size in the size range of about 10 microns or less, advantageously with most in the range of about 2 microns or less. The bioactive composition of the invention may be produced and/or obtained by various methods, including but not limited to: a) a modified low-temperature sol-gel process producing inorganic composites; b) a modified low-temperature sol-gel process producing organically modified silicates; c) chemically or physically modifying naturally occurring analogs of the binary oxide material obtained from chemical supply companies, i.e.; calcium silicate from wollastonite; and d) the amorphous precipitation of inorganic materials. IN THE DRAWINGS Figure 1 is a scanning electron photomicrograph or "SEM" (10 KV X 3200 magnification) of a sample of calcified tissues, an untreated dentin disc surface incubated in sterile saliva. Figure 2 is a SEM of a treated dentin disc surface (prepared the same way as the untreated disc surface) after being treated (or repaired) by materials made by the precipitation-based process producing inorganic particles. Figure 3 is a scanning electron photomicrograph (20 KV X 6000 magnification) of an untreated dentin disc surface. Figure 4 shows a similarly prepared dentin disc surface after being treated with materials made by the dilute precipitation process. Figure 5 is a scanning electron photomicrograph (10 KV X 1700 magnification) of an untreated dentin disc surface. Figure 6 shows a similarly prepared dentin disc surface after being treated with materials made in Example 2 - modified sol-gel ORMOSILS process. Figure 7 is a scanning electron photomicrograph (10 KV X 1600 magnification) of the control untreated dentin disc sample. Figure 8 (10KV x 1700) shows a similarly prepared dentin disc surface after being treated with materials prepared in Example 4 - modified sol gel - base catalyzed reaction. DETAILED DESCRIPTION OF THE INVENTION The inventors have found a surprisingly low cost and convenient-to-manufacture bioactive materials that restore defects in calcified tissues, including the alleviation of pain associated with hypersensitive teeth. The inventors have also found a bioactive glass that is conducive to mineralization by varying the processing parameters and the wet chemistry of the materials. Bioactive Particulate Composition. Current bioactive mateπals, including bioactive glass, often rely on a complicated list of oxides, including P2O5, Na 2 0, B2O 3 , K 2 O, MgO, AI 2 O 3 , Tι0 2 , Ta 2 θ 5 and fluoπde salts, including CaF 2 We surpπsingly found that nucleating crystal growth can be initiated by utilizing only a binary oxide system based essentially exclusively on Si, O and Ca +2 The bioactive mateπals prepared in accordance with the present invention, a CaO-Si0 2 containing inorganic/organic hybπd or inorganic composite deπved from naturally occumng analogs, are capable of depositing a layer of apatite or other calcium-containing mineral, similar in composition and crystal nity to those mineral phases that predominate m normal dentin once the mateπal's surface is placed in contact with blood, saliva or simulated body fluid. We have also found that by retaining bioactivity while removing these species from the composition, we can lower the cost of the mateπal, allow greater freedom m processing and eliminate possible organoleptic and toxicological problems. The simplicity of the chemical composition of the present invention, which not only aids in facilitating processing and loweπng costs, but also allows flexibility to tailor the composition to the intended application. We found a mateπal tailored to nucleating calcium-containing mineral for the treatment of a specific problem, tooth sensitivity, and removed excipients that may be a necessity for biological activity when used as an osteo-inductive mateπal elsewhere in the body Unlike most currently-used bioactive mateπals that act m part by encouraging m-growth and tissue formation via osteoblasts and other hard tissue cells, the mateπals of the present invention surpπsingly do not require cellular adjuvants Additionally, the compositions in accordance with the present invention are able to induce the precipitation of apatite and calcium-containing mineral from simulated body fluids without the aid of bone morphogenic proteins, dentin phosphoprotems, odontoblasts or their host cells, or a defined collagen matπx The ability to induce mineralization without the need for organic adjuvants optimizes this mateπal for use in the oral environment In addition, many of these currently-used mateπals need highly specialized properties such as porosity and mechanical strength suited to their specific clinical use For example, Li, Clark and Hench showed that there is a minimum rate of hydroxyapatite formation which is necessary to be an effective mineralizing agent and that this rate is a function of both composition and microstructure (Li et. al , 'An Investigation of Bioactive Glass Powders by Sol-Gel Processing, J. of Applied Biomateπals, Vol. 2, 231-239 [1991]). The inventors have surpπsingly found that the mateπals of the present invention do not need the porosity and mechanical strength of the pπor art bioglasses to work in dental applications, so that when lodged in the tubules they initiate the formation of calcium-containing mineral inside the dentinal tubules. The inventors have also surpπsingly found that the bioactive mateπals of the present invention to be bioactive well beyond the compositional boundary for melt-den ved glasses of the pπor art with a less porous microstructure Although it is not necessary, other oxides including P 2 0 5 , Na 2 0, B 2 0 3 , K 2 0, MgO, A1 2 0 3 , Tι0 2 , Ta 2 θ5 and fluoπde salts, including CaF 2 , as commonly called for in the compositions m the pπor art, can be used m conjunction with the binary oxide system of the present invention. A preferred and exemplary application for the bioactive mateπals of the present invention is in the treatment of dentin hypersensitivity In microscopic examination of exposed sensitive dentin surfaces show that clear differences exist between sensitive and non-sensitive regions of dentin Sensitive dentin is permeated by open tubules. In contrast, non-sensitive tooth surfaces are characteπzed by tubules that are sealed off from the external environment by naturally occumng mineral deposits or coveπngs We found that once bound to the dentin surface or lodged within the tubule, the amorphous mateπal of the present invention initiates πucleation and mineralization stimulated by the salivary environment, sealing the tubule from the external stimuli by depositing a layer of apatite or other calcium contaimng mineral, similar to native tooth mineral in composition and crystalhnity Camers suitable for use with the composition for use m the treatment of dentin hypersensitivity are preferably hydroxy c mateπals such as water, polyols and mixtures thereof. Polyols, sometimes referred to as humectants, include glycerol, sorbitol, propylene glycol, xyhtol, polypropylene glycol, polyethylene glycol, hydrogenated com syrup and mixtures thereof. Particularly preferred as the carrier is a liquid mixture of 3 - 30% water, 0- 90% glycerol and 0 - 80% sorbitol. Generally the amount of the carrier will range from about 25 to 99.9% by weight, preferably from about 70 - 95% by weight. When the compositions of this invention are in the form of a toothpaste or gel for the treatment of dentin hypersensitivity, there will typically be included a natural or synthetic thickening agent in an amount from about 0.1 - 10%, preferably about 0.5 - 5% by weight. Thickeners may include hydroxypropyl methylcellulose, hydroyethyl cellulose, sodium carboxymethylcellulose, xanthan gum, tragacanth gum, karaya gum, gum arabic, Irish moss, starch, alginates and carrageenans. Surfactants are normally included in compositions for the treatment of dentin hypersensitivity. These surfactants may be of the anionic, nonionic, cationic or amphoteric type. Most preferred are sodium lauryl sulfate, sodium dodecylbenzene sulfonate and sodium laurylsarcosinate. Surfactants are usually present in an amount from about 0.5 - 80% by weight. For anti-caries protection, a source of fluoride ion will normally be present in the oral compositions. Fluoride sources include sodium fluoride, potassium fluoride, stannous monofluorophosphate, calcium fluoride, stannous fluoride and sodium monoflurophosphate. These sources should release from about 25 - 3500 ppm fluoride ion. The anti-caries agents will be present in an amount from about 0.05 - 3.0% by weight, preferably about 0.5 - 1.0%. Flavors that are usually present in oral care compositions are those based on oils of spearmint and peppermint. Examples of other flavoring agents include menthol, clove, wintergreen, eucalyptus and aniseed. Flavors may range in concentration from about 0.1 - 5.0%. Sweetening agents such as saccharin, sodium cyclamate, aspartame, sucrose and the like may be included at levels from about 0.1 - 5.0% by weight. Other additives may be incorporated including preservatives, silicones, other synthetic or natural polymers, anti-gingivitis actives, anti-tartar agents, whitening agents and other desirable products often found in a conventional toothpaste such as baking soda and peroxide. It is also advantageous that these materials are compatible with conventional desensitizing agents such as potassium nitrate, potassium chloride and potassium bicarbonate, as well as novel desensitizing agents such as those described in U.S. Patent No. 5,589,159. Numerous vehicles are present for the oral delivery of active agents in the treatment of dentin hypersensitivity, including but not limited to, pastes, gels, rinses, powders, gums, dental floss, slurries and solutions and although each is not described it is within the scope of the present invention. In compositions for use as bone implants or to form implantable surfaces, other materials can be added to the bioactive materials of the present invention including antibiotics, structural fibers and / or polymer resins, depending on the application. Preparation. It is an advantage of the current invention that the materials are easy to fabricate eliminating the costly and time consuming processing procedures involved with the melt-derived and sol-gel derived bioactive materials in the prior art. The bioactive materials of the present invention can be prepared by several processes and with several chemical modifications designed to uniquely tailor the composition to a specific application because of advanced processing and by tailoring the wet chemistry to enhance the material's bioactivity. The processes of the present invention can be used to prepare the bioactive materials of the present invention, which are based exclusively on Si, O and Ca +2 . 1. Modified Sol-Gel Process Producing Inorganic Glasses. Conventional sol-gel synthesis of glass is achieved by combining a metal alkoxide precursor with water and a catalyst with consequent "polymerization" of the metal alkoxide species and the production of a gel followed by the following steps: 1) heat sintering at temperatures often between 600 - 900 degrees centigrade, 2) calcination heat treatments, and often 3) supercritical drying to maintain the integrity of the nanostructured material. Unlike glasses prepared by the melt-derived process, glasses prepared by the sol-gel process ("sol-gel-derived glasses") maintain bioactivity for compositions up to pure silica gels. A particular characteristic of sol-gel processing is the production of microporous materials. As a result two advantageous consequences follow: 1 ) The high surface area associated with the porous calcium-containing silica gel leads to a rapid increase in the concentration of Ca +2 ions in the surrounding solution, and 2) The texture produced by the sol-gel process results in a porous gel layer even with the reduced ion exchange and dissolution rates as the Si0 2 content increases. These two factors are thought to be responsible for the high rate of mineral formation and for the extension of the compositional bioactivity range in the sol-gel derived glasses. Studies have shown that sol-gel prepared silica, unlike melt-derived glass, has a highly hydrolyzed silica surface which facilitates and initiates apatite nucleation from metastable simulated body fluids. The inventors have been able to surprisingly incorporate the favorable porosity and increased surface area associated with sol-gel derived materials in a "modified sol-gel process" that does not require the additional processing steps of heat sintering, calcination heat treatments, and supercritical drying to maintain the integrity of the nanostructured material, which steps are not easily adaptable to the production but necessary to achieve "a correctly formed sol-gel glass." In this modified sol-gel process of the invention to produce inorganic materials, a calcium alkoxide precursor, tetraethoxysilane (TEOS) and optionally triethylphosphate (TEP), is hydrolyzed under acidic conditions and low temperatures to form a gel. Other appropriate ingredients will also be apparent to those of ordinary skills in the art. The process can also be performed using alkaline conditions for the hydrolysis reaction. This has been found to control both the morphology and size of the powders produced. 2. Modified Sol-Gel Process Producing Organically Modified Silicates ("Ormosils"). In this Ormosils process, a hydrolysis reaction as described in the modified sol-gel process is first used to prepare organic / inorganic hybrid materials, followed by the formation of an inorganic network. During the build-up of this network, appropriately functionalized organic moieties also undergo a condensation reaction and are incorporated in the forming network. In this second manifestation, organic/inorganic hybπds are prepared from silanol terminated poly(dimethylsιloxane) [PDMS] and tetraethoxysilane [TEOS] precursors (a source of silicon) as descπbed by Hu and MacKenzie (Journal of Mateπal Science, 27:4415-4420 [1992]). The final particle size, surface modifications and control of reactivity can be controlled by varying processing parameters, specifically by the choice of catalyst and the pH of the system. It is advantageous that the flexibility of the mateπals can be controlled by changing the mixing ratios of the organic and inorganic components. If Ca(II) and other key species for bioactivity are introduced into the organic/inorganic hybπd mateπal, these composites will exhibit both flexibility and bioactivity. The ability to control particle size, surface morphology, flexibility and bioactivity of the mateπals of the present invention enables the unique tailoπng of these mateπals to their intended application. 3. Using / modifying Natural Minerals (Commercially Available Silicates). The bioactive agents of the present invention may be prepared by utilizing or modifying naturally occurring analogs of the inorganically produced material, i.e., calcium silicate obtained from wollastonite. Commercial soluble silicates have the general formula: M 2 θ (Si0 2 ) m (H 2 0) n where M is an alkali metal and m and n are the number of moles of Si0 2 and H 2 0, respectively, per mole of M 2 0. The composition of commercial silicates is typically described by the weight ratio of Siθ 2 to M2O. These soluble silicates have many uses, the largest and most rapidly growing of which aπses from the ability to serve as a source of reactive pπmary silica species. The ability of this inorganic mateπal to readily form the reactive silica species is the key to its ability to nucleate calcium-containing mineral. The rate of dissolution for soluble silicates depends on the glass ratio, solids concentration, production temperature, pressure, panicle size and overall surface area. Dissolution typically occurs in a two-step mechanism that involves ion exchange and network breakdown. Ion exchange: ≡Si-OCa + H 2 0 <= ≡Si-OH + Ca +2 + OH ' Network breakdown: ≡Si-O-Sii≡ + OH => ≡Si-O " + HO-Si≡ As supplied, calcium silicate may not conform to the specifications or possess the functionality desired and certain modifications, i.e. surface alterations, particle size adjustment, and/or other necessary treatments may be implemented. When first received, the commercial material from Alfa Aesar was moderately active in our evaluation. However, after ball-milling the material to a mean size of 5μm, the material provided enhanced bioactivity. Naturally occurring calcium silicate obtained from wollastonite has shown bioactivity in our in vitro evaluations. These materials including calcium silicate, (meta (CaSi0 3 ), ortho (Ca 2 Si0 ), etc, can be obtained commercially through major chemical supply companies such as Alfa Aesar and Aldrich. 4. Precipitation-Based Process Producing Inorganic Particles. The fourth manifestation of the present invention is our most preferred process. In this precipitation-based process, the sol-gel procedure is modified to a precipitation method to remove the difficult processing parameters inherent with a sol-gel (or the "modified sol-gel" method) simply by precipitating an amorphous oxide and not going through the 'sol-gel' stage. In this process, inorganic particles are produced as previously described in the sol-gel process, by the controlled hydrolysis of TEOS and resulting condensation incorporating reactive ions donated from a calcium source, either a calcium salt such as calcium nitrate or an alkoxide derivative such as calcium methoxide in a precipitation method. The inorganic particles contain specific agents, ions, polymers or colloidal particles that render the materials of the present invention bioactive. In a traditional sol-gel (or our "modified" sol-gel) procedure, silicate gels are often synthesized by hydrolyzing monomeric tetrafunctional alkoxide precursors employing a mineral acid (e.g., HC1) or base (e.g., NH 3 ) as a catalyst. At the functional group level, three reactions are generally used to describe the sol-gel process: 1. Hydrolysis ≡Si-OR + H 2 0 = ≡Si-OH + ROH Esterification 2. Alcohol Condensation ≡Si-OR + HO - Sis <_ =Sι-0-Sι≡ + ROH Alcoholysis 3 Hydrolysis ≡Si-O- Sis + H 2 0 o ≡Si-OH + HO - Sι≡ Water Condensation where R is an alkyl group, C H jt+ y. The hydrolysis reaction replaces alkoxide groups (OR) with hydroxyl groups (OH) Subsequent condensation reactions involving the silanol groups produce siloxane bonds (Si-O-Si) plus the by-products alcohol (ROH) or water. Under most conditions, condensation commences before hydrolysis is complete. Because water and alkoxysilane are immiscible, a mutual solvent such as alcohol is normally used as a homogenizing agent m a sol-gel process. However, gels can be prepared from silicon alkoxide-water mixtures without added solvent, since the alcohol produced as the by-product of the hydrolysis reaction is sufficient to homogenize the imtially phase separated system. We have found that, if after the hydrolysis step, a reactive species, le: calcium nitrate, is added to the mixing vessel, the silica will precipitate because of the surface charge destabi zation within the sol. This precipitated mateπal that when tested in our model, surpnsmgly achieves supeπor results as a dentmal tubule occluder and, as judged by scanning electron microscopy ("SEM"), a supeπor dentin mineralizing agent as shown in the Figures, compaπng two calcified tissue samples or two dentin disc surfaces, one untreated (Figure 1) and one treated (Figure 2) with the mateπals prepared by this preferred process. In the most preferred method of the precipitation process, a "dilute precipitation process," an amorphous CaO-Sιθ2 inorganic mateπal is produced by the reaction of a dilute silicate source, i.e., aqueous potassium or sodium silicate solutions with a soluble calcium source as previously descπbed This procedure allows for the manipulation of both the composition of the final precipitate as well as physical characteπstics such as the final size or concentration Mateπals prepared by this dilute process are found to work very well in both aqueous dispersion and prototype dentifrice compositions as confirmed by hydraulic conductance tests and SEMs as shown in Figures 3 and 4 (two similarly prepared dentin surfaces, untreated and treated). Not wishing to be bound by theory, it is postulated in our invention, Ca(II) and other key species for bioactivity may be introduced into the forming silica sol resulting in a particulate hybrid material. In general, the stabilization of colloids by electrostatic repulsion is successfully described by the DLVO theory (the Derjaguin, Landau, Verivery and Overbeak theory describing the stability suspensions) well-known to colloid chemists. Silica does not conform to the DLVO theory because it is apparently stabilized by a layer of adsorbed water that prevents coagulation even at the isoelectric point. The addition of cations to the aqueous silica sol may reduce the degree of hydration and destabilize the silica. Allen and Matijevic (J. Colloid and Interface Sci, 31[3] 1969 287 - 296, 33[3] (1970) 420 - 429, 35 [1] (1971) 66-76) showed that adding a salt to the sol would produce ion exchange in the following manner: sSi-OH + M 2 → ≡Si-OM (z - I)+ + H + where M* * is an unhydrolyzed cation of charge z. Since the silanol groups are the adsorption sites for water, the removal of SiOH by ion exchange reduces the amount of hydration and lessens the stability of the colloid. The reaction precipitates a Ca(II)-containing silicate that can be easily removed from solution by filtration and resuspended in a vehicle suitable for application as a dental material. The remaining solutions may be reused facilitating both cost and production of the material. In the precipitation-based process of the present invention, sodium silicate [CAS# 1344-09-8] may be substituted as the silicon source. The addition of an appropriate amount of a soluble calcium source, i.e. Ca(N0 3 ) 2 , to a solution of sodium silicate precipitates a calcium-containing silicate whose composition is similar to previously described materials. The amount of Ca * to be added to the silicate solution was determined by both stoichiometric calculations and experimental design. The precipitation-based process of the present invention has several advantages over other processes. This process produces no by-products that may contaminate the final product after incoφoration into a standard dentrifice, i.e., ethanol or residual TEOS. In addition, the chemistry of the reaction in this process is such that particle size can more easily be controlled during the reaction, thus eliminating the need for further size reduction and milling steps which are not only costly but also may increase the chance for contamination of the final product. Further, the reaction requires only two ingredients, sodium silicate and the appropriate calcium source. Both of these chemicals are readily available from common chemical supply companies and are relatively inexpensive compared with the other processes. In this process, the material, once precipitated and separated from the remaining solvent, does not require any special processing, including washing, drying at high heat or supercritical drying; nor does the material involve a calcination procedure or organic solvent extraction. It is additionally preferred, although not critical, that in the process, the precipitated material is chemically controlled to produce amoφhous material of sufficiently small particle size range as to not require additional milling, grinding or size reduction. Thus, one avoids the possibility of sample contamination and additional processing costs. The preferred process of chemical reactions performed under ambient conditions produces a highly porous, calcium-containing silica similar to bioactive glass prepared by the traditional sol- gel process in biological activity. This inorganic material does not resemble the heat-sintered bioactive glass material currently available in other ways. It is most preferred that the reaction produces a calcium-containing oxide material based solely on a two component system, CaO-Si0 2 ; and that the reaction occurs at room temperature in solvents familiar to the health care field as well as safe and well-known to production facilities personnel. The bioactive materials prepared in the examples that follow are suitable for various applications, including but not limited to: a) the enhancement of bone cell attachment and activity when the materials are placed in a tissue culture medium for the repair of osseous defects, such that when inoculated with cells, bone tissue formation is initiated; b) implant compositions to fill cavities in bone and canals in teeth; c) oral care compositions for use in remineralizing teeth; c) oral care compositions for periodontal osseous defect repair; and oral care compositions for use in the treatment of dentin hypersensitivity. Osseous defects contemplated include, but are not limited to, the following: cystic defect repairs, benign and malignant tumor sites upon resection, bone loss, fracture repair sites including delayed- or non-union sites, joint repair sites, osteoporosis- related defects, and periodontal defects. Examples The following examples are provided for illustrative puφoses only, and changes or alterations may be made that are not disclosed therein. In examples tailored toward the treatment of dentin hypersensitivity, experiments were conducted using a modified in vitro model of dentin sensitivity described in U.S. Patent 5,270,031. In this method, intact human third molars free of caries or restorations are sectioned peφendicular to the long axis of the tooth with a metallurgical saw into sections approximately 1 mm. thick. Sections containing dentin and free of enamel are retained for testing. These sections are then etched with an EDTA (ethylenediamine tetraacetic acid) solution to remove the smear layer. The disk is mounted in a split chamber device as reported in Joumal of Dental Research 57:187 (1978). This special leak proof chamber is connected to a pressurized fluid reservoir containing a tissue culture fluid intended to mimic the osmolarity of human body fluid. By using a mixture of pressurized N 2 and C0 2 gas, the fluid can be maintained at physiological pH. The apparatus includes a glass capillary tube mounted on a ruler or other measuring instrument. An air bubble is injected into the glass capillary tube and by measuring the displacement of the bubble as a function of time the fluid flow through the dentin disk can be measured. It has been reported that fluid actually flows out of dentin tubules from the interior of a normal tooth. Following measurements of the baseline fluid flow in the dentin disk, the experimental mixture, dentifrice, gel or mouthwash is applied to the external disk surface in a manner that would mimic its use clinically. After a defined application period, the experimental material is rinsed off and post-application (lpacute) hydraulic conductance is measured. In this fashion various experimental materials both alone and as components of final products can be tested for the ability to obstruct fluid flow in the dental tubules. The percent flow reduction induced by application of the experimental material can then be calculated. For all examples given, saliva incubation was necessary to achieve flow reductions. For the puφose of studying the mineralizing potential of novel agents, an extended time period is necessary as well as a modification of the standard dentin hydraulic conductance measurement. After initial post-application measurements are taken, the split chamber is removed from the apparatus and, a reservoir of sterile filtered human saliva is attached to the chamber allowing the fluid to bathe the treated occlusal dentin surface. The exterior of the chamber is wrapped in parafilm to avoid contact with air and resulting fluid loss. The unit is then placed in an incubator kept at body temperature. After a sufficient time, the unit is removed, the saliva reservoir is discarded and the split chamber is re-attached to the hydraulic conductance apparatus. At this time, dentin fluid flow can be re-measured and the experimental material can be re-applied. In the examples that follow, the flow reductions reported were obtained after three treatments and 40 hours of saliva incubation. In the examples that follow as throughout this specification and claims, all temperatures are in degrees centigrade and all parts and percentages are by weight unless otherwise indicated. Example 1 (Modifed sol-gel - ORMOSILS). The starting materials were reagent grade calcium nitrate, TEOS (tetraethoxysilane), PDMS (polydimethylsiloxane). Reagent grade 2-propanol and tetrahydrofuran were used as solvents and HC1 was used as a catalyst. TEOS (lOg) and silanol terminated PDMS (5.9g) were mixed with a mixture of 2-propanol (4.8ml) and tetrahydrofuran (3.2ml). This solution was denoted Solution A. An appropriate amount of calcium nitrate (0.42 g) was dissolved in distilled water and HC1 solution (35%). 2-propanol (7.0ml) was added to the solution while stirring; this became Solution B. This recipe is based upon the synthesis of bioactive Ormosils outlined by K. Tsuru et al, Mat. Res. Soc. Symp. Proc. Vol. 435, p. 404, 1996. Solutions A and B were mixed and subsequently refluxed while stirring at 80°C for 30 minutes. After the reflux, the mixture was quenched to 25°C with iced water, cast into containers and allowed to gel under ambient conditions. After the gelation, flat, irregularly shaped shards were milled to sizes in the range of about 10 microns or less, with most in the range of about 2 microns or less. Milling can be performed by several methods including but not limited to ball-milling, air-impact milling, MICROS superfine milling, rotary cutter milling, hammer milling, and cage milling. This material, now of known particle size, was dispersed in glycerin and tested in our in vitro model as a slurry at various concentrations. In the current embodiment of the invention, repeated applications of a 20% slurry showed good dentin fluid flow reducing ability as seen in Table 1. Table 1 : Percent Flow Reduction with Glycerin Dispersions of an Organic/Inorganic Composite Material It should be noted that the cycle of repeated applications and filtered saliva incubations did not produce significant fluid flow reductions when non-bioactive materials were evaluated. In addition, the integrity of an untreated dentin surface and a placebo treated dentin surface, exposed to this cyclic method, was confirmed by SEM. No occlusion was observed when a finely ground pumice treated dentin surface was examined in this manner. Similarly, the moφhology of untreated dentin surfaces that were exposed to the cyclic sterile saliva treatments alone, as in this mineralization model, did not significantly differ from freshly cut and prepared dentin specimens after SEM evaluation. Example 2 (Modifed sol-gel - ORMOSILS). The second embodiment of the present invention is prepared from the same precursors as in Example 1 and processed in a similar manner. However, in this example we significantly increased the total amount of calcium nitrate (1.25 g), the calcium ion source, in the material. Although this increase reduced the unusual mechanical strength and pliability of the original material, the resulting composition showed better dentin fluid flow reducin ability (Table 2) at various concentrations in glycerin dispersions. Table 2: Percent Flow Reduction with Glycerin Dispersion of an Organic/Inorganic Material with high calcium ion incoφoration This material resulted in a more rapid and complete occlusion than did the material described in Example 1 , as demonstrated in Figure 6 which showing a dentin disc surface after being treated with materials prepared by this modified sol-gel ORMOSILS process. Figure 5 shows a similarly prepared dentin surface that is not treated. Example 3. (Modified sol-gel - Inorganic materials). A completely inorganic material is produced by a sol-gel process under ambient conditions. The material uses precursors similar to Example 2, but PDMS and tetrahydrofuran were omitted. The 2-propanol is a solvent for TEOS and not used in this example. The TEOS is simply a source of silicon in this example. The procedure is the same as in Examples 1 and 2. However, the product of this example contains only calcium, silicon, and oxygen. Although this material has poor mechanical strength, it is very effective as a tubule occluding agent. In addition, the absence of PDMS and tetrahydrofuran is favorable for the toxicological profile. Table 3: Percent Flow Reduction with Glycerin Dispersion of an Sol-Gel Derived Inorganic Composite Example 4 (Modified sol-gel process - Base catalyzed reaction). A base-catalyzed sol-gel process is employed to control the shape and size of the resulting particulate suspensions. The starting materials are as follows: 50 ml TEOS, 75 ml Ethanol, 70 ml water, 0.575g ammonium hydroxide (30%) and 3.6g calcium nitrate. For these reactions, ammonia is used as a catalyst causing the formation of spherical particles. This procedure is based upon a 1968 Stδbber and Fink article, "Controlled Growth of Monodisperse Silica Spheres in the Micron Size Range" (Journal of Colloid and Interface Science, 26, 62-69, 1968). The resulting particulate composition proved to be a very effective mineralizing agent (Table 4). Scanning electron micrographs showed complete coverage of the dentinal tubules inferred to be due to mineralization of the tubule orifice in samples treated with the composite material. Untreated samples are marked by numerous patent tubules indicative of hypersensitive regions of the tooth. In addition, our ability to chemically control shape and moφhology of the particulate material makes this example the easiest to formulate and process. In addition, elemental analysis of this material confirms large amounts of Ca +2 and Si in a ratio conducive to bioactivity. Figures 7 and 6 compare two similarly prepared dentin surfaces, one untreated and one treated with the composition of Example 8. Table 4: Percent Flow Reduction with Glycerin Dispersion of an Inorganic Composite Example 5 (Naturally occuπng minerals) A calcium ort/io-si cate [(CaO) 2 -Sι0 2 , FW- 0.088 - 0 044mm (172.24, - 325 mesh US)] deπved from the mineral Wollastonite from Alfa Aesar was used. Published studies have indicated that pseudowollastonite, synthesized at 1500°C for 2 hours from a stoichiometπc mixture of calcium carbonate and silica, is bioactive m a simulated body fluid environment (De Aza PN. et al., Bioactivity of Pseudowollastonite m Human Saliva, Journal of Dentistry, 27 (1999), 107-113) As supplied from Alfa Aesar, the particle size was unsuitable for such an application and the mateπal was ball-milled in glycerol to a mean particle size favorable to mineralization of a microporous substrate As shown in Table 5, the resulting particulate composition proved to be an effective mineralizing agent Table 5 Percent Flow Reduction with Dispersions of Calcium ort io-Si cate deπved from Wollastonite Example 6 (Precipitation-Based Process Producing Inorgamc Particles) A precipitate was prepared using commercially available calcium nitrate and sodium silicate solution. To 72.8 wt % of a 40% sodium silicate solution, 27.2 wt. % of a 75% calcium tetrahydrate solution in deionized water was added, with the mixer running at high speed to provide maximum agitation to the sodium silicate solution. Precipitation occurred immediately The precipitate (Calcium Silicate, Inorgamc Precipitate or "CSIP") was dπed at 60°C for 40 hours. In example 6, the CSIP particle size was reduced by milling (e.g., using an air impact mill, simple ball mill, etc.). However, it is possible to alter the particle size by alteπng the pH or diluting the sodium silicate solution while maintaining the high calcium nitrate concentration of loweπng the final particle size of the precipitate. Supeπor tubule occluding efficacy was judged both by hydraulic conductance and SEMs as shown Figures 3-4 (untreated and treated with the composition of Example 6) Example 7 A toothpaste composition which further aids in the alleviation of pain when used on hypersensitive teeth was prepared using the following formulation. The desensitizing agent in this example can be any of the aforementioned Ormosil, inorganic composites, synthetically derived inorganic material, or sol-gel derived materials, or sodium silicate derived materials in a suitable composition. The weight percent given for each ingredient is based on a value of 100% for the total formulation. Example 8. A second toothpaste formulation was prepared comprising a desensitizing agent consisting of either an Ormosil, inorganic composite, synthetically derived inorganic material, or sol-gel derived materials. The weight percentage basis and the process of preparation are the same as that set forth in previous examples. Example 9 A third toothpaste formulation was prepared as a two-phase system. This toothpaste may be in either one tube where each phase is kept separate by a septum dividing the package, it may be in a dual chamber pump that dispenses each phase simultaneously, or it may be packaged in separate tubes meant to be applied sequentially. The active ingredient may also be formulated into a variety of dental rinse formulations designed to alleviate tooth sensitivity. An extremely fine particle size is needed to ensure homogeneous dispersion of either the Ormosil, inorganic composites, synthetically derived inorganic material, sol-gel derived materials or naturally-derived calcium silicate from mineral precursors. This can be accomplished by either chemical modification or mechanical milling the of material. Example 10. Formulations of a Medicinal-Type Mouthwash and Fluoride Oral Rinse are as follows in weight percent (adjusted to pH 6):

Claims: Claims 1. A composition for restoring defects in calcified tissues comprising an amoφhous bioactive particulate material consisting essentially of calcium, silicon and oxygen. 2. A composition according to claim 1 wherein said amoφhous material is selected from the group consisting of: a) a reaction product of an organic silicate source and a source of calcium; b) a calcium containing hydrolysis product of tetraethylorthosilicate; c) a calcium containing silica sol-gel; d) a binary calcium oxide and silicate precipitated material; e) a synthetic analog of a naturally occurring wollastonite-like calcium silicate; and f) a precipitated reaction product of a soluble calcium source and a silicate solution. 3. A composition according to claim 2 wherein said amoφhous material is a precipitated reaction product of a soluble calcium source and a dilute silicate source selected from an aqueous potassium solution or a sodium silicate solution. 4. A composition according to any of claims 1 to 3, in which the said material has a particle size in the range of about 10 microns or less. 5. A composition according to claim 4, in which the material mostly comprises particles in the size range of about 2 microns or less. 6. A composition according to any of claims 1 to 5 in a dental product to alleviate tooth sensitivity due to the presence of exposed dentinal surface and open dentinal tubules, by causing the formation of calcium containing mineral within dentinal tubules of the sensitive teeth. 7. A composition according to claim 6 comprising a carrier and about 1-25% of said amoφhous material. 8. A composition according to claim 7 wherein the earner compπses 0-30% water, 0-90% glycerol and 0-80% sorbitol. 9. A composition according to claim 7 or claim 8 wherein said amoφhous mateπal is about 5-10% of the composition and the composition is a dentifrice. 10. Use of an amoφhous bioactive particulate consisting essentially of calcium, silicon and oxygen, in the preparation of product to restore defects m calcified tissues. 11. A use according to claim 10 in the preparation of product to alleviate tooth sensitivity due to the presence of exposed dentmal surface and open dentinal tubules by causing the formation of calcium containing mineral within dentinal tubules of the sensitive teeth. 12. A use according to claim 10 or claim 11 wherein said amoφhous mateπal is selected from the group consisting of: a) a reaction product of an organic silicate source and a source of calcium; b) a calcium containing hydrolysis product of tetraethylorthosilicate; c) a calcium containing silica sol-gel; d) a binary calcium oxide and silicate precipitated mateπal, e) a synthetic analog of a naturally occurπng wollastonite- ke calcium silicate; and f) a precipitated reaction product of a soluble calcium source and a silicate solution. 13. An amoφhous bioactive particulate consisting essentially of calcium, silicon and oxygen, for restoπng defects in calcified tissues. 14 An amoφhous bioactive particulate according to claim 13, for use in the desensitizing of teeth. 15. An amoφhous bioactive particulate according to claim 13 or claim 14 selected from the group consisting of: a) a reaction product of an organic silicate source and a source of calcium; b) a calcium containing hydrolysis product of tetraethylorthosilicate; c) a calcium containing silica sol-gel; d) a binary calcium oxide and silicate precipitated material; e) a synthetic analog of a naturally occurring wollastonite-like calcium silicate; and f) a precipitated reaction product of a soluble calcium source and a silicate solution. 16. A process for producing an amoφhous bioactive particulate for restoring defects in calcified tissues, said process consisting essentially of adding a sufficient amount of a solution of a soluble calcium source to a solution of a silicate or silicate precursor to cause a precipitation of calcium-containing silicate particuiates. 17. A process according to claim 16, wherein said soluble calcium source is selected from the group consisting of calcium salts and alkoxide derivatives and mixtures thereof. 18. A process according to claim 16 or claim 17, wherein said solution of a silicate or silicate precursor is a dilute silicate source selected from the group of aqueous potassium solution and aqueous sodium silicate solution. 19. A process for the production of an amoφhous bioactive particulate for the treatment of sensitive teeth, said process consisting essentially of combining a metal alkoxide with water and a catalyst and polymerization of a silicate ester for the production of said amoφhous bioactive particulate. 20. A process according to claim 19, wherein said metal alkoxide is prepared from tetraethoxysilane and silanol terminated poly(dimethylsiloxane).

Inventors: DODD GREGORY P, HALECKY ALAN A, MARKOWITZ KENNETH J, DODD, GREGORY, P., HALECKY, ALAN, A., MARKOWITZ, KENNETH, J.
Applicants: BLOCK DRUG CO, WHALLEY KEVIN, BLOCK DRUG COMPANY, INC., WHALLEY, KEVIN
Application year: 2000
Main classification: A61K7
IPC codes: A61K6, A61K8, A61K33, A61L27, A61P1, A61P19, A61Q11