Patent Description:
The clinical application of calcium phosphate systems in medicine has been known since the identification of the bone mineral composition but autologous and allogeneic bone implants have been and still are preferentially used, with undeniable advantages such as their osteoinductivity, bioactivity, biocompatibility, osteoconductivity, as well as a high degree of ossteointegration with the surrounding tissue after healing of the damaged sites. The above characteristics have not been observed in such complexity in synthetic types of calcium phosphate biomaterials especially in the case of ossteoinduction and ossteointegration. The advantage of synthetic biomaterials is that there is no immunogenic response and possible transmission of diseases from the donor. Moreover, there are no complications associated with autologous transplantation with the removal of biological material from another part of the body. The widespread use of calcium phosphate biomaterials for commercial purposes in the form of coatings, hydroxyapatite ceramic blocks and fixtures begins in the <NUM>, while porous ceramic scaffolds were only developed at the end of the 20th century (<NPL>; <NPL>). Calcium phosphate biocements (CPC) are biocompatible, osteoinductive and osteoconductive materials depending on composition and they are used for reconstruction and regeneration of bone tissues in the body, because due to their easy application in the form of a moldable paste, they are predestined for perfect filling of various types of defects in bone tissues in the cranio-maxillofacial region. (<NPL>; <NPL>), in dental medicine and orthopaedics (<NPL>).

In terms of addressing osteochondral defects, the patented systems are mainly oriented towards the preparation of cellular types of biopolymer 3D-substrates with seeded autologous stem cells or chondrocytes in the form of, for example, devitalized allo- or xeno-grafts of cartilage containing cartilage growth factors (<CIT>, <CIT>), formation of synthetic matrix on substrates by chondrocytes in-vitro (<CIT>), injectable biopolymer microspheres (<CIT>), cellulose hydrogels (<CIT>), photocomposites based on chitosan/collagen blends (<CIT>), silk-based multistructural porous systems with fibrin adhesive bonded layers (<CIT>), composite calcium phosphate/biopolymer films (<CIT>) and fibrous bilayer scaffolds (<CIT>), composite implants made of decellularized cartilage and demineralized bone (MD1177), calcium phosphate/collagen sponges (<CIT>) or multiphase scaffolds (<CIT>), gradient porous scaffolds containing PLGA microparticles and hydrogels (<CIT>), devitalized micronized extracellular matrix (<CIT>), multi-layered collagen/hyaluronic acid blends (<CIT>), double-walled structures with hydroxyapatite and fibroin (<CIT>) chemically bonded with chitosan (<CIT>), ceramic structural bi-layered system with different porous layers (<CIT>), collagen/hydroxyapatite multilayer structures (<CIT>). In addition, papers have been published and patented on the application of calcium phosphate biocements demonstrating successful healing of bone and osteochondral defects in vivo (SK288818; <NPL>).

CPCs are most commonly represented by two- and multi-component systems, with individual components undergoing dissolution and hydrolysis, which changes the nature of the environment (e.g., pH, increase in ion concentration) with subsequent supersaturation of surroundings in relation to the final product of precipitation, most commonly hydraxyapatite (HAP) (or its calcium-deficient forms, CDHAP) (<NPL>). Hydroxyapatite self-setting biocement types are mostly composed of tetracalcium phosphate/monetite (CaHPO<NUM>) (TTCP/DCPA or brushite (CaHPO<NUM>. <NUM><NUM>O) mixtures or contain a high proportion of tricalcium phosphate (mostly the high-temperature α-form), and after mixing with the liquid component (phosphate solutions), a gradual transformation of the phases to CDHAPs takes place and setting of the whole system occurs (<NPL>; <NPL>). The physicochemical properties in relation to the setting process of TTCP/DCPA-based CPCs are significantly influenced by the Ca/P ratio of the CPC, the addition of organic additives (carboxylic acids, alginates, phytoleic acid, chitosan, etc.), various inorganic fillers (inert oxides - SiO<NUM>, ZrO<NUM>, silicon nitrides, whiskers, etc.), as well as by the addition of HAP nanocrystalline particles (<NPL>; <NPL>; <NPL>; <NPL>; <NPL>). The amorphous to nanocrystalline nature of HAP particles in CPCs influences the in vitro behaviour of biologically active bone tissue cells (osteoclasts, osteoblasts) and promotes cell proliferation and growth (<NPL>; <NPL>; <NPL>; <NPL>; <NPL>). A small (~<NUM>% w/w) addition of different types of organic substances (e.g., soluble starch, cyanuric acid, cyclodextrin, dextran, polyvinylpyrrolidone) to TCP cement increased the stability of the cement paste after immersion to solutions (<NPL>). The formation of the TCP/gelatine composite significantly enhanced the compressive strength of the cement (<NPL>).

The bioactivity of the CPC cement system and especially the resorbability can be increased by the addition of calcium sulphates as an additional component of the cement, however, the amount must be optimized to obtain suitable physicochemical and mechanical properties, as they alone have poor mechanical properties and a high resorption rate (<NPL>). Calcium sulphate (CS) is added to cements to increase the porosity of cements up to <NUM>% w/w (<NPL>; <NPL>; <NPL>) and improving the mechanical properties of the original CPC mixture (<NPL>), which were reduced with the rise in porosity after dissolution of CS (<NPL>). The addition of calcium sulphate hemihydrate actively affected the transformation of TTCPM cement mixture and reduced the rapid pH rise after cement paste preparation (<NPL>; <NPL>).

The bioactivity of CPCs in relation to the formation of new bone tissue can be positively influenced by the use of biologically active agents such as bone morphogenetic proteins and growth factors dosed in very small amounts (<NPL>), which led to a significant increase in bone formation and neovascularisation in the rabbit femoral defect model (<NPL>). Similarly, αTCP cement acquired osteoinductivity by adding nanogram quantities of BMP2 to the cement porous granules, from which it was gradually released after implantation into the rabbit (<NPL>). The aforementioned findings demonstrate the effective and powerful influence of specific substances mostly produced by cells on the complete formation of bone tissue. The problem with these substances, mostly proteins, is their high cost, the possibility of a decrease in their activity after adsorption on the biomaterial due to a change in their structural configuration as well as the necessity to kept storage conditions (mostly -<NUM>) after processing in order to preserve their biological characteristics. The solution may lie in the search for suitable inexpensive natural products with a relatively wide range of biological characteristics covering the stimulation of osteoinductive, osteoconductive as well as anti-inflammatory or antimicrobial properties of CPCs. Examples include Aloe vera containing acermanic acid, aloein, which increased mesenchymal stem cell (MSC) proliferation, ALP activity and promoted bone tissue growth (<NPL>). In addition to its anti-inflammatory and antimicrobial properties, curcumin has a positive effect on the proliferation of osteoblasts and suppresses the resorption of bone tissue (<NPL>). Allicin as an active component of garlic, protects osteoblasts from oxidative stress and apoptosis (<NPL>). Green tea is another interesting example of the effective impact of a natural product on osteoporosis, inhibition of bone resorption and growth of osteoblastic activity (<NPL>; <NPL>).

In terms of bone tissue, bone waxes containing originally beeswax, almond oil and acetylsalicylic acid are very often applied and are characterized by a homeostatic effect in orthopaedic (total knee replacement) (<NPL>) or neurological surgeries because they have excellent adhesion to bone and are hydrophobic, acting as an impermeable, non-resorbable membrane that does not leak blood after their application (<NPL>). Their disadvantages are significant side effects associated with bacterial infection, formation of fibrous tissue, insufficient formation of new bone tissue, development of a strong inflammatory reaction, etc. The above shortcomings have been addressed by the development of new bone waxes, where beeswax has been replaced by synthetic products such as PEG, acid esters, fatty acids in conjunction with cellulose hydrogels, chitosan, gelatine, and to increase bioactivity, CPCs are added and gradual degradation of the wax was demonstrated (<NPL>). Another type of natural product is honey, long known for its antimicrobial properties and effective effect in healing skin wounds. Lowers ambient pH, promotes vascular growth factor production, epithelialization and granulation of tissue as well as reduces swelling (<NPL>). The importance of honey for regenerative medicine applications of bone defects is still underestimated, and there are virtually no papers analysing the properties of calcium phosphate systems in ceramic or CPC form containing honey. Honey containing polyphenolic compounds with strong antioxidant properties suppresses the oxidative stress of cells, thus effectively counteracting osteoporotic changes and the decline in osteoblast activity. In animal models, it has been shown that the addition of honey to the diet increased calcium absorption into bone tissue in the acute phases and promoted biomechanical bone strength after long-term use (<NPL>). Application of honey to artificially created defects in rat mandible demonstrated significant growth of well-mineralized bone tissue already in the early stages of healing (<NPL>). Analysis of bone tissue in a rat radius defect confirmed improved morphology of newly formed tissue in a honey/HAP composite (<NPL>). The results of the analysis of the influence of honey on the precipitation of calcium phosphate in solutions revealed its inhibitory but also promotional effect depending on the type of honey used (<NPL>).

<CIT> discloses a cement system composed of a powder component comprising a calcium phosphate mixture of microcrystalline tetracalcium phosphate and a monetite with a Ca/ P mole ratio in the range of <NUM> to <NUM>; with an average particle size of <NUM> to <NUM> and a liquid component comprising a solution containing up to <NUM>% w/w of sodium dihydrogen phosphate or sodium hydrogen phosphate.

The subject matter of the invention is the composition of a composite biocement system designed for reconstruction and regeneration of hard tissue, cartilage and osteochondral defects. The biocement system according to the invention is as defined in claims <NUM> and <NUM>.

Wherein the honey in the liquid component is preferably in the range of <NUM> to <NUM>% w/w, most preferably <NUM> to <NUM>% w/w. The calcium phosphate component of the biocement is characterized by a Ca/P mole ratio corresponding to stoichiometric or calcium-deficient hydroxyapatite in the range of <NUM>-<NUM>. The basic powdered tetracalcium phosphate component of the biocement mixture is synthesized by high temperature reaction at temperatures above <NUM> of a homogeneous powdered mixture of various calcium phosphates preferably calcium hydrogen phosphate (CaHPO<NUM>), calcium hydrogen phosphate dihydrate (CaHPO<NUM>. <NUM><NUM>O), calcium dihydrogen phosphate (Ca(H<NUM>PO<NUM>)<NUM>), calcium pyrophosphate (Ca<NUM>P<NUM>O<NUM>) as well as the diammonium hydrogen phosphate ((NH<NUM>)<NUM>HPO<NUM>) with calcium carbonate (CaCO<NUM>), calcium oxide (CaO), calcium hydroxide (Ca(OH)<NUM>) or other calcium salts which decompose at temperatures up to <NUM>. The obtained tetracalcium phosphate is further refined by grinding in a ball mill, usually for <NUM>-<NUM> minutes, so that the resulting powder contains particles with a particle size of <<NUM>. The final tetracalcium phosphate/monetite cement mixture is prepared in situ by reacting the synthesized milled tetracalcium phosphate with a dilute solution of orthophosphoric acid in a pure organic solvent or a mixture of solvents miscible with water preferably in ethanol, methanol, acetone, or mixtures thereof, the orthophosphoric acid being added in a well-defined amount to give a total Ca/P molar ratio in the range of <NUM>-<NUM>. The tetracalcium forphate/monetite mixture containing calcium sulphate hemihydrate is prepared from powdered tetracalcium phosphate by in situ reaction with a mixture of orthophosphoric acid and sulfuric acid in a pure organic solvent or a mixture of solvents miscible with water and preferably in ethanol, methanol, acetone or mixtures thereof, so that the final product of the cement setting is stoichiometric hydroxyapatite or calcium-deficient forms thereof with a total Ca/P molar ratio in the range <NUM>-<NUM>. Sulphuric acid is added in such an amount that the calcium sulphate content of the final cement mixture is preferably up to <NUM>% w/w.

The powdered component of the composite biocement system may also be prepared directly by dry (without liquid medium) or wet homogenization of the powdered mixture of calcium phosphate precursors in a mill with a total Ca/P mole ratio in the range of <NUM>-<NUM>, or by mechanical mixing of suspensions in water, ethanol, methanol, acetone, or mixtures thereof, which may be absolute, i.e., completely anhydrous, or may contain a well-defined amount of water in the range of up to <NUM> % v/v.

The dried powdered mixture does not need to be further treated in any way, and the final composite biocement system in the form of a cement paste is obtained by mixing the prepared powdered calcium phosphate mixture with a solution containing up to <NUM>% w/w of sodium dihydrogen phosphate or sodium hydrogen phosphate or potassium dihydrogen phosphate or mixture thereof, preferably a <NUM>-<NUM>% solution of sodium dihydrogen phosphate, sodium hydrogen phosphate, potassium dihydrogen phosphate, or a mixture thereof, and up to <NUM> % w/w of honey. The ratio of the powder to liquid component may be in the range of <NUM>-<NUM>, preferably <NUM>/ml. The setting times of biocement systems prepared according to the described method are in the range of <NUM>-<NUM> minutes, depending on the composition of the cement mixture, the grinding time, the concentration and the amount of liquid. The compressive strength of hardened cements is in the range up to <NUM> MPa, which corresponds to the strength characteristics of bone tissue. The advantage of biocement mixtures prepared by in situ reaction is a significant reduction of the pH value in the initial stages of setting below <NUM> as well as the addition of honey, which reduces the intensity of irritation, inflammatory reaction and stress factors of the surrounding tissues after application. The biocements are easy to apply at the site of the defect immediately after preparation of the cement paste and surgical repair of the defect site, have satisfactory viscosity and consistency for filling bone defects as well as high resistance to disintegration in aqueous solutions after only <NUM> minutes from the addition of the liquid component. In the case of cement with the addition of calcium sulphate hemihydrate, there is an increase in the concentration of calcium ions resulting from its dissolution and transformation during the setting of the cement paste. The results of in vivo experiments confirmed the formation of bone tissue of comparable morphology and composition to the original bone tissue and, in the case of an osteochondral defect, formation of a continuous hyaline cartilage with a zone structure within <NUM> weeks without signs of inflammatory processes at the site of the defect.

Any references in the description to methods of treatment refer to the products of the present invention for use in a method for treatment.

A composite biocement system composed of a calcium phosphate mixture containing powdered microcrystalline tetracalcium phosphate with an average particle size <<NUM> and nanocrystalline monetite. The content of the individual components in the biocement corresponds to a Ca/P mole ratio of <NUM>. The liquid component of the cement contained <NUM>% w/w of NaH<NUM>PO<NUM> and <NUM>% w/w of honey. The setting time was <NUM> and the compressive strength reached <NUM> MPa.

A composite biocement system composed of a calcium phosphate mixture containing powdered microcrystalline tetracalcium phosphate with an average particle size <<NUM> and nanocrystalline monetite. The content of the individual components in the biocement corresponds to a Ca/P mole ratio of <NUM>. The liquid component of the cement contained <NUM>% w/w of NaH<NUM>PO<NUM> and <NUM>% w/w of honey.

A composite biocement system composed of <NUM>% w/w of tricalcium phosphate and calcium phosphate mixture with a Ca/P mole ratio of <NUM> containing powdered microcrystalline tetracalcium phosphate with an average particle size <<NUM> and monetite. The liquid component of the cement contained <NUM>% w/w of NaH<NUM>PO<NUM> and <NUM>% w/w of honey.

A composite biocement system consisting of tricalcium phosphate with an average particle size < <NUM>. The liquid component of the cement contained <NUM>% w/w of NaH<NUM>PO<NUM> and <NUM>% w/w of honey. The powder-to-liquid ratio was <NUM>. The setting time was <NUM> and the compressive strength reached <NUM> MPa.

A composite biocement system consisting of tricalcium phosphate with an average particle size <<NUM>. The liquid component of the cement contained <NUM>% w/w of honey dissolved in water. The powder-to-liquid ratio was <NUM>.

A composite biocement system consisting of tricalcium phosphate with an average particle size <<NUM>. The liquid component of the cement contained <NUM>% w/w of KH<NUM>PO<NUM>, <NUM>% w/w of NaH<NUM>PO<NUM> and <NUM>% w/w of honey.

A composite biocement system consisting of tricalcium phosphate with an average particle size <<NUM>. The liquid component of the cement contained <NUM>% w/w of NaH<NUM>PO<NUM> and <NUM>% w/w of honey.

A composite biocement system composed of <NUM>% w/w of calcium sulphate hemihydrate and calcium phosphate mixture with a Ca/P mole ratio of <NUM> containing powdered microcrystalline tetracalcium phosphate with an average particle size <<NUM> and monetite. The liquid component of the cement contained <NUM>% w/w of NaH<NUM>PO<NUM> and <NUM>% w/w of honey.

A composite biocement system composed of <NUM>% w/w of tricalcium phosphate and calcium phosphate mixture with a Ca/P mole ratio of <NUM> containing powdered microcrystalline tetracalcium phosphate with an average particle size <<NUM> and monetite. The liquid component of the cement contained <NUM>% w/w of KH<NUM>PO<NUM> and <NUM>% w/w of honey.

A composite biocement system composed of <NUM>% w/w of tricalcium phosphate, <NUM> % w/w of calcium sulphate hemihydrate and calcium phosphate mixture with a Ca/P mole ratio of <NUM> containing powdered microcrystalline tetracalcium phosphate with an average particle size <<NUM> and monetite. The liquid component of the cement contained <NUM>% w/w of NaH<NUM>PO<NUM> and <NUM>% w/w of honey.

A composite biocement system composed of a calcium phosphate mixture containing powdered microcrystalline tetracalcium phosphate with an average particle size <<NUM> and monetite. The content of the individual components in the biocement corresponds to a Ca/P mole ratio of <NUM>. The liquid component of the cement contained <NUM>% w/w of NaH<NUM>PO<NUM> and <NUM>% w/w of honey.

A composite biocement system composed of <NUM>% w/w of tricalcium phosphate, <NUM> % w/w of hydroxyapatite and calcium phosphate mixture with a Ca/P mole ratio of <NUM> containing powdered microcrystalline tetracalcium phosphate with an average particle size <<NUM> and monetite. The liquid component of the cement contained <NUM>% w/w of NaH<NUM>PO<NUM> and <NUM>% w/w of honey.

The initial powdered tetracalcium phosphate phase was prepared by annealing a mixture of CaHPO<NUM>. <NUM><NUM>O and CaCO<NUM> so that the Ca/P mole ratio was equal to <NUM> at <NUM> for <NUM> hours in air. The tetracalcium phosphate phase was crushed and milled in a ball mill for <NUM> minutes and the resulting powder had an average particle size equal to <NUM>. The calcium phosphate cement system was prepared in situ by milling powdered tetracalcium phosphate in a planetary ball mill for <NUM> minutes in an amount of orthophosphoric acid solution (diluted <NUM>:<NUM> by volume in absolute ethanol) that corresponds to the resulting Ca/P molar ratio of Examples <NUM> and <NUM>. The result was a homogeneous powder precursor mixture containing microcrystalline tetracalcium phosphate and nanocrystalline monetite, to which a liquid component is added containing <NUM>% w/w of NaH<NUM>PO<NUM> and <NUM>% w/w of honey. The powder-to-liquid ratio was <NUM>.

The initial powdered tetracalcium phosphate phase was prepared by mixing CaHPO<NUM>. <NUM><NUM>O and CaCO<NUM> so that the Ca/P mole ratio was equal to <NUM>. The mixture was then annealed at <NUM> for <NUM> hours in air. The tetracalcium phosphate phase was crushed, milled in a ball mill for <NUM> minutes and sieved (Mesh <NUM>). Subsequently, the powdered tetracalcium phosphate was placed in a grinding vessel and milled in a planetary ball mill in a solution of a mixture of orthophosphoric acid and sulfuric acid (mixture diluted <NUM>:<NUM> in <NUM>% ethanol) for <NUM> minutes. The amount of orthophosphoric acid and sulfuric acid was added so that the total Ca/P mole ratio in the suspension was equal to <NUM> and the calcium sulphate content was <NUM>% w/w. The resulting suspension was poured onto a watch glass and dried at <NUM> for <NUM> hours, and the resulting powdered biocement system was mixed with a liquid cement component containing <NUM>% w/w of NaH<NUM>PO<NUM> and <NUM>% w/w of honey. The powder-to-liquid ratio was <NUM>.

The powdered tetracalcium phosphate/monetite/calcium sulphate hemihydrate cement mixture prepared according to Example <NUM> was mixed with tricalcium phosphate so that the content of the tricalcium phosphate component in the biocement system was <NUM>% w/w, and the resulting powdered biocement system was mixed with a liquid cement component comprising <NUM>% w/w of KH<NUM>PO<NUM> and <NUM>% w/w of honey. The powder-to-liquid ratio was <NUM>.

The powdered tetracalcium phosphate phase was prepared by mixing CaHPO<NUM> and CaCO<NUM> so that the Ca/P mole ratio was equal to <NUM>. The mixture was then annealed at <NUM> for <NUM> hours in air. The tricalcium phosphate phase was crushed, milled in a ball mill for <NUM> minutes and sieved (Mesh <NUM>). The powdered tetracalcium phosphate/monetite calcium phosphate mixture prepared according to Examples <NUM> and <NUM> was mixed with tricalcium phosphate so that the content of the tricalcium phosphate component in the biocement system was <NUM>% w/w, and the resulting powdered biocement system was mixed with a liquid cement component comprising <NUM>% w/w of KH<NUM>PO<NUM> and <NUM>% w/w of honey. The powder-to-liquid ratio was <NUM>.

The powdered tetracalcium phosphate/monetite calcium phosphate mixture prepared according to Example <NUM> was mixed with tricalcium phosphate and hydroxyapatite so that the content of the tricalcium phosphate component in the biocement system was <NUM>% w/w and the content of the hydroxyapatite component was <NUM>% w/w, and the resulting powdered biocement system was mixed with a liquid cement component comprising <NUM>% w/w of KH<NUM>PO<NUM> and <NUM>% w/w of honey. The powder-to-liquid ratio was <NUM>.

The tricalcium phosphate phase was crushed, milled in a ball mill for <NUM> minutes, sieved (Mesh <NUM>) and mixed with a liquid cement component containing <NUM>% w/w of KH<NUM>PO<NUM>, <NUM>% w/w of Na<NUM>HPO<NUM> and <NUM>% w/w of honey. The powder-to-liquid ratio was <NUM>.

A subchondral articular cartilage defect in the left knee joint was induced in female sheep aged <NUM>-<NUM> years under general anaesthesia. The incision was made from the left lateral side, from the medial patellar ligament distal to the tibial tuberosity. The knee joint was made accessible over the weight-bearing surface of the medial femoral condyle. The subcutaneous ligament and superficial fascia were disrupted. In flexion of the knee joint and partial luxation of the knee bone, the Osteochondral autograft transfer system (Arthrex) kit was used to induce an articular cartilage defect at a predefined site of the distal femoral epiphysis (distal femoral epiphysis - 1x left medial condyle), with a diameter of <NUM> and a depth of <NUM>. The site of the formed defect was subsequently filled with calcium phosphate biocement according to Example <NUM>.

A subchondral articular cartilage defect in the left knee joint was induced in female sheep aged <NUM>-<NUM> years under general anaesthesia. The incision was made from the left lateral side, from the medial patellar ligament distal to the tibial tuberosity. The knee joint was made accessible over the weight-bearing surface of the medial femoral condyle. The subcutaneous ligament and superficial fascia were disrupted. In flexion of the knee joint and partial luxation of the knee bone, the Osteochondral autograft transfer system (Arthrex) kit was used to induce an articular cartilage defect at a predefined site of the distal femoral epiphysis (distal femoral epiphysis - 1x left medial condyle) and the proximal part of the tibia, with a diameter of <NUM> and a depth of <NUM>. The site of the formed defect was subsequently filled with calcium phosphate biocement according to Example <NUM>. Sampling was performed <NUM> months after implantation of the tested biomaterial. Histological analyses showed no inflammatory process at the implantation site, morphological evaluation demonstrated the formation of hyaline cartilage tissue with a zonal structure, and histochemical analysis confirmed the presence of glycosaminoglycans in active chondrocytes. Macrographs and radiographs confirmed almost complete healing of the tibial and subchondral defect (<FIG>).

The advantage of biocement mixtures prepared by in situ reaction is a significant reduction of the pH value in the initial stages of solidification below <NUM> as well as the addition of honey, reduces the intensity of irritation, inflammatory reaction and stress factors of the surrounding tissues after application. An in vitro comparison of the total antioxidant capacity of extracts from the composite biocement system and cement without the addition of honey demonstrated more than twice the antioxidant capacity of the composite systems. Similarly, statistically significantly increased (<NUM>-<NUM>%) osteogenic gene expression of osteocalcin, osteonectin and osteopontin was identified after in vitro culture of osteoblasts in extracts of composite systems compared to extracts of pure cement.

Claim 1:
A composite biocement system characterized in that it is composed of a powder component comprising a calcium phosphate mixture of microcrystalline tetracalcium phosphate with an average particle size of less than <NUM> and a monetite with a Ca/P mole ratio in the range of <NUM> to <NUM> and a liquid component comprising honey dissolved in a solution containing up to <NUM>% w/w of sodium dihydrogen phosphate or sodium hydrogen phosphate or potassium dihydrogen phosphate or mixture thereof.