Preparation and fully compounded stock for use in medical or dental applications, medical or dental product and use and preparation thereof

Abstract

According to the invention, a preparation is described which contains at least one calcium compound selected from the group consisting of calcium phosphates, calcium fluorides and calcium fluorophosphates and hydroxyl derivatives and carbonate derivatives of these calcium salts, calcium hydroxides and calcium oxides precipitated using at least one protein component selected from proteins and protein hydrolysates, and at least one crosslinking agent for the protein component and/or non-set cement.

Claims

1. A preparation comprising: at least one calcium compound selected from the group consisting of: calcium phosphates, calcium fluorides and calcium fluorophosphates and hydroxyl derivatives and carbonate derivatives of these calcium salts, calcium hydroxides and calcium oxides, which is precipitated using at least one protein component selected from proteins and protein hydrolysates, at least one cross-linking agent for the at least one protein component, wherein the at least one cross-linking agent is selected from the group consisting of: transglutaminase, sortase A, tyrosinase, laccase, peroxidase, lysiloxidase, amine oxidase, glutaraldehyde, (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide/N-hydroxysuccinimide, Genipin, caffeic acid, hexamethylene diisocyanate, proanthocyanidin and formaldehyde, and an unset cement.

2. The preparation according to claim 1, wherein the unset cement is selected from the group consisting of calcium silicate cement, calcium phosphate cement, and mixtures of calcium silicate cement and calcium phosphate cement.

3. The preparation according to claim 1, wherein the at least one calcium compound is selected from the group consisting of: Ca(H.sub.2PO.sub.4).sub.2.Math.xH.sub.2O, wherein x is an integer of from 0 to 6, CaHPO.sub.4.Math.xH.sub.2O, wherein x is an integer of from 0 to 6, Ca.sub.8(HPO.sub.4).sub.2(PO.sub.4).sub.4.Math.5H.sub.2O, Ca.sub.3(PO.sub.4).sub.2, Ca.sub.10(PO.sub.4).sub.6(OH).sub.2, Ca.sub.10(PO.sub.4).sub.6F.sub.2, (Ca.sub.10-aM.sub.a) [(PO.sub.4).sub.6-bY.sub.b] [(OH).sub.2-cX.sub.c] wherein M=Na.sup.+, Sr.sup.2+, Mg.sup.2+, Ba.sup.2+, Pb.sup.2+; YHPO.sub.4.sup.2, CO.sub.3.sup.2; XF.sup., Cl.sup., H.sub.2O; wherein a is an integer of from 0 to 10, b is an integer of from 0 to 6, and c is an integer of from 0 to 2, Ca(OH).sub.2 and CaO.

4. The preparation according to any one of claim 1, wherein the at least one protein component is selected from the group consisting of: collagen, keratin, wheat protein, rice protein, soy protein, almond protein and hydrolysates thereof.

5. The preparation according to claim 1, wherein the at least one protein component is gelatin.

6. The preparation according to claim 1, wherein a content of the at least one cross-linking agent in the preparation is more than 0 to 25% by mass, based on a total mass of the preparation.

7. The preparation according to claim 1, wherein a content of the at least one cross-linking agent in the preparation is more than 0 to 10% by mass based on a total mass of the preparation.

8. The preparation according to claim 1, wherein a content of the at least one cross-linking agent in the preparation is more than 0 to 4% by mass, based on a total mass of the preparation.

9. The preparation according to claim 1, and further comprising at least one pigment selected from the group consisting of oxides, hydroxides or oxyhydroxides of iron, titanium or zinc and any mixtures of oxides, hydroxides or oxyhydroxides of iron, titanium or zinc.

10. The preparation according to claim 1, and further comprising at least one water-soluble fluoride.

11. The preparation according to claim 10, wherein the at least one water-soluble fluoride is NH.sub.4F, KF or NaF.

12. The preparation according to claim 10, wherein a content of the at least one water-soluble fluoride in the preparation is more than 0 to 10% by mass, based on a total mass of the preparation.

13. The preparation according to claim 10, wherein a content of the at least one water-soluble fluoride in the preparation is more than 0 to 5% by mass, based on a total mass of the preparation.

14. The preparation according to claim 1, and further comprising casein.

15. The preparation according to claim 14, wherein a content of the casein in the preparation is more than 0 to 30% by mass, based on a total mass of the preparation.

16. The preparation according to claim 14, wherein a content of the casein in the preparation is more than 0 to 15% by mass, based on a total mass of the preparation.

17. The preparation according to claim 14, wherein a content of the casein in the preparation is more than 0 to 5% by mass, based on a total mass of the preparation.

Description

EXAMPLES

(1) Unless otherwise stated, percentages given refer to mass %. Furthermore, PO.sub.4 is understood to mean phosphate (PO.sub.43-).

(2) 1. Production of a Composite or Composite Compound

(3) 1.1 Production of Calcium Phosphate-Protein Component Composites:

(4) Examples for the preparation of calcium phosphate-protein component composites, i.e. calcium phosphate precipitated using a protein component, are given below. As an example, gelatine is used as the protein component, but other protein components, such as those disclosed above, may also be used.

Example 1

Apatite-Gelatine Composite

(5) More than 0 to 25 g, especially 3 g of gelatine was dissolved in 500 ml of H.sub.2O at 45 C. and then cooled to 25 C. Dissolving gelatine (general: protein component) can generally be done in a temperature range above 0 C. to about 70 C. 45 C. was found to be optimal to quickly obtain a uniformly dissolved gelatine solution. Subsequently, 22.05 g CaCl.sub.2-2H.sub.2O (0.15 mol) was added, and generally the amount of CaCl.sub.2-2H.sub.2O can be varied throughout the solution range of CaCl.sub.2 as long as the molar ratio of Ca to PO.sub.4 is adjusted to 1.5 to 1 to 1.67 to 1. Then, the pH of the solution was adjusted to pH 9, although the pH generally can be from 7 to 11 to finally obtain apatite. The solution was stirred for about 30 minutes, causing the solute ions to attach to the gelatine. Thus, a sort of a pre-structurization took place.

(6) In parallel, a second solution was prepared from 12.42 g NaH.sub.2PO.sub.4H.sub.2O (90 mmol) in 250 ml H.sub.2O. Again, the molar ratio of Ca to PO.sub.4 was variable in the range of from 1.5 to 1 to 1.67 to 1. This second solution was titrated to the first solution at pH kept constant by 1 M NaOH, at about 3 ml/min (0.1 ml to about 20 ml/min is possible). After completion of the addition, stirring was continued at constant pH for another 24 h (another 2 h to 365 days is generally possible), then centrifuged and washed four times with 55 C. H.sub.2O. Samples were then either stored under refrigeration for direct use, lyophilized, or dried at 50 C. for hardness measurement.

(7) The particles obtained herein, as long as they were freeze-dried, had a platelet-like structure with a thickness of a few nanometers and an extension of less than 100 nm. A white powder was obtained.

(8) When the platelets were dried at elevated temperature as well as cross-linked, these platelets collapsed and adhered to each other, forming the desired dentin-like structure. In this process, a solid tooth-like material was obtained. Without crosslinking of the protein component, the material obtained had a hardness of approx. 25-30 HV0.3. When crosslinked with transglutaminase and casein, a hardness of up to 72 HV0.3 could be achieved.

(9) The protein content could be varied over a very wide range.

(10) In each case, the hardness measurement was carried out according to Vickers HV0.3: see Metallic materialsVickers hardness testPart 1: Test method (ISO 6507-1:2018); German version EN ISO 6507-1:2018.

Example 2

Octacalcium Phosphate (OCP)-Gelatine Composite

(11) The synthesis of the OCP-gelatine composites was carried out according to the same principle as the synthesis of the apatite composite. Except the order of calcium and phosphate addition and molar ratio thereof. Thus, in the standard procedure herein, 3 g of gelatine (also variable from more than 0 to about 25 g) was dissolved in 500 ml of H.sub.2O at 45 C. and then cooled to room temperature. Then, 12.42 g of NaH.sub.2PO.sub.4H.sub.2O (90 mmol) was added (the amount of NaH.sub.2PO.sub.4H.sub.2O can be varied throughout the solution range of NaH.sub.2PO.sub.4H.sub.2O as long as the molar ratio of Ca to PO.sub.4 is always adjusted to 1.33 to 1), the pH was adjusted to 7 (the pH can be varied in the range of 5 to 7.5 to finally obtain octacalcium phosphate), and stirred for 30 minutes for pre-structurization, resulting in attachment of the dissolved ions to the gelatine.

(12) In parallel, a second solution consisting of 17.64 g CaCl.sub.2-2H.sub.2O (0.12 mol) in 250 ml H.sub.2O was prepared (as described above, variable in the ratio to PO.sub.4) and, with pH 7 being constant by addition of 1 M NaOH (pH can be varied in the range of pH 5 to 7.5), the calcium chloride solution was added to the phosphate solution at 3 ml/min (variable from 0.1 to 20 ml/min). The solution was then either directly centrifuged as well as washed or stirred for 24 h for maturation (variable from 1 h to 365 days) and then centrifuged/washed as well as freeze-dried as desired. Herein, when freeze-drying, particles having a thickness of a few nm and an extension of several hundred nm were obtained, which were in the form of a white powder.

(13) When drying by elevated temperature, no hard material was obtained herein.

(14) The protein content could be adjusted very well.

Example 3

Brushite-Gelatine Composite

(15) 3 g of gelatine (variable from more than 0 to 25 g) was dissolved in 500 ml of H.sub.2O at 45 C. and then cooled to 25 C. (temperatures between 0 and 70 C. are also possible), followed by addition of 17.64 g of CaCl.sub.2-2H.sub.2O (0, 12 mol) (the amount of CaCl.sub.2-2H.sub.2O can be varied throughout the solution range of CaCl.sub.2 as long as the molar ratio of Ca to PO.sub.4 is always adjusted to 1 to 1) followed by pH adjustment to pH 5 (can be varied in the range of pH 2 to 5 to finally obtain brushite). The solution was then stirred for half an hour for pre-structurization.

(16) In parallel, a second solution of 16.598 g NaH.sub.2PO.sub.4H.sub.2O (0.12 mol) in 250 ml H.sub.2O was prepared (as described above, variable in relation to Ca), which was titrated at 3 ml/min (variable from 0.1 to 20 ml/min) at pH kept constant by 1 M NaOH after the end of the prestructuring phase. The order of Ca or PO.sub.4 addition may also be reversed, i.e. NaH.sub.2PO.sub.4H.sub.2O may also be added and CaCl.sub.2 titrated. After completion of the addition, stirring was continued at constant pH for another 24 h (variable between 1 h and 365 days), then centrifuged and washed four times with 55 C. H.sub.2O. Samples were then either stored under refrigeration or freeze-dried.

(17) Larger platelets with a thickness of several hundred nm and an extension of 10 to 100 m were obtained. When freeze-drying as well as drying by elevated temperature, a powder was obtained.

(18) The gelatine content was significantly lower. Only contents up to 5 mass % were obtained.

Example 4

Amorphous Calcium Phosphate (ACP)-Gelatine Composite

(19) 3 g of gelatine (variable from more than 0 to 25 g) was dissolved in 500 ml of H.sub.2O at 45 C. and then cooled to 25 C. (other temperatures between 0 and 70 C. are also possible), followed by the addition of 24.51 g of CaCl.sub.2-2H.sub.2O (0, 167 mol) (the amount of CaCl.sub.2-2H.sub.2O can be varied throughout the solution range of CaCl.sub.2) followed by pH adjustment to pH 10 (can be varied in the range of pH 2 to 12 to finally obtain ACP) with subsequent half-hour stirring for pre-structurization.

(20) In parallel, a second solution of 13.799 g NaH.sub.2PO.sub.4H.sub.2O (0.10 mol) in 250 ml H.sub.2O was prepared (the ratio of Ca to PO.sub.4 here is variable in molar ratio from 1.2 to 1 to 2.2 to 1), which was titrated at 6 ml/min (variable from 4 to 30 ml/min) at pH kept constant by 1 M NaOH after the end of the prestructuring phase. The order of Ca or PO.sub.4 addition may also be reversed, i.e. NaH.sub.2PO.sub.4H.sub.2O may also be added and CaCl.sub.2 titrated. After completion of the addition, samples were directly centrifuged and washed four times with 55 C. H.sub.2O. Samples were then either lyophilized or dried at 50 C. for hardness measurement.

(21) Partially spherical structures which were very poorly defined were obtained, yielding a white powder upon freeze-drying.

(22) During normal drying, transformation towards apatite occurred, resulting in a solid tooth-like material.

(23) The gelatine content could be adjusted very well. Contents from 0 to 30 mass % were obtained.

(24) 2. Production of Medical or Dental Products Using Biomimetic Dental Cements/Biomimetic Filling Materials as an Example

(25) General Production Process when Employing Calcium Phosphate Cements:

(26) For the production of biomimetic dental cements according to the invention, different calcium compounds selected from calcium phosphate phases were used in combination with the previously described calcium phosphate-gelatine composites to achieve recrystallization towards an apatite phase having a biomimetic structure by combining calcium-rich phases and calcium-poor phases.

(27) Parallel to curing of the cement materials initiated by the inorganic phase, an additional curing step was carried out herein by crosslinking the gelatine (protein component) contained with various crosslinkers, in particular with transglutaminase in combination with casein. This second additional crosslinking step, in addition to the setting reaction by recrystallization, significantly contributed to the good mechanical, chemical and biological properties as well as the excellent long-term stability of the dental filling material.

(28) FIG. 2 reviews the achieved hardnesses of composites crosslinked with different crosslinking agents.

(29) In a typical experiment, various calcium salts having different calcium contents as well as phosphate-containing salts (all salts listed in Table 1 can be used in all possible compositions) were mixed such that a molar ratio of calcium to phosphate of 1.5 to 1 to 1.67 to 1 was adjusted, with the ratio of 1.67 to 1 being preferred, since this corresponds to the ratio in pure apatite.

(30) The salts were used in this process by grinding them into various particle sizes between 100 m and 1 nm in order to change the reaction rates and properties of the materials. In addition to the calcium phosphate salts, a proportion of previously synthesized calcium phosphate composite was added and then crosslinked with a crosslinker during curing to form an organic supporting network. In addition, it was always possible to add further additives to improve the structure, for example by defoaming or adding fluoride, or to increase the radiopacity, for example by adding radiopaque materials, and to adapt the product to the tooth color using a dye.

(31) TABLE-US-00001 TABLE 1 Overview of compounds for producing calcium phosphate-gelatine composites and dental cements from calcium phosphate cements and composite. Possible reactants Molecular formula Ca/PO.sub.4 ratio Phosphoric acid H3PO.sub.4 0 (100% PO.sub.4) Salt of the phosphoric acid AxByPO.sub.4 0 (100% PO.sub.4) Monocalciumphosphate Ca(H2PO.sub.4)H.sub.2O 0.5 monohydrate (MCPM) Monocalciumphosphate Ca(H2PO.sub.4)xH.sub.2O 0.5 hydrate Monocalciumphosphate Ca(H2PO.sub.4) 0.5 anhydrous (MCPA) Brushite CaHPO.sub.4H.sub.2O 1 (dicalciumphosphate dihydrate) Brushite-gelatine composite CaHPO.sub.4H.sub.2O + gelatine 1 Monetite CaHPO.sub.4 1 (dicalciumphosphate anhydrous) Octacalciumphosphate Ca8(HPO.sub.4)2(PO.sub.4)45H.sub.2O 1.33 (OCP) Octacalciumphosphate- Ca8(HPO.sub.4)2(PO.sub.4)45H.sub.2O + Gelatine 1.33 Gelatine-Composite -/- Tricalciumphosphate Ca.sub.3(PO.sub.4).sub.2 1.5 (TCP) Hydroxyapatite (HAP) Ca.sub.10(PO.sub.4).sub.6(OH).sub.2 1.667 Apatite (Ca, Ba, Pb, Sr, etc.)5(PO.sub.4, 1.33-1.667 HPO.sub.4, CO3)3(F, Cl, OH) (or more than 1.67) Fluorapatite (FAP) Ca.sub.10(PO.sub.4).sub.6F.sub.2 1.667 HAP/FAP-Gelatine- Ca.sub.10(PO.sub.4).sub.6(OH).sub.2 + gelatine 1.667 Composite Tetracalciumphosphate Ca4(PO.sub.4)2O 2 (TTCP) Amorphous CaxHy(PO.sub.4)znH.sub.2O, 1.2-2.2 calciumphosphates (ACP) n = 3-4.5; ACP-Gelatine Composite 1.2-2.2 Calciumhydroxide Ca(OH)2 100% Ca Calciumoxide CaO 100% Ca Calcium salts Ca(AxBy) 100% Ca

Example 5

Production of Biomimetic Dental Cements Based on Wet Apatite-Gelatine Composites

(32) For producing a biomimetic dental cement using wet apatite-gelatine composites, the procedure was to dry mix calcium-containing as well as phosphate-containing salts in a Ca/PO.sub.4 molar ratio of 1.667 to 1 or to grind them together. In addition to the calcium or phosphate salts, a proportion of fluoride-containing salts or carbonate-containing salts could be added to obtain fluorapatite-substituted or carbonate-substituted apatite. After mixing all the dry materials, an amount of wet apatite-gelatine composite (in different compositions in terms of protein and water content) and, if necessary, water were then added to obtain a processable paste. Addition of water initiated the reaction between the salts used towards the apatite and curing of the entire material.

(33) In parallel with the addition of the composite material, the use of gelatine-crosslinking agents (see FIG. 2) in varying proportions was able to achieve additional curing of the composite material, thus further improving the material properties.

(34) A specific example thereof given the following is the formation of the biomimetic dental cement based on wet apatite-gelatine composites:

(35) 0.7 g -tricalcium phosphate (2.26 mmol) having an average particle size in the range of 1-10 m was dry-triturated together with 0.042 g CaO (0.75 mmol) as well as 0.0252 g NaF (0.6 mmol). In parallel, a second mixture consisting of 0.233 g wet apatite-gelatine composite (water content 75%; protein content 5%; apatite content 20%) together with 0.017 g transglutaminase (for example ultrafiltration from Ajinomoto Activa WM) as well as 0.0085 g casein and 0.15 ml H.sub.2O was mixed for 20 seconds in the universal mixer. Both phases were then mixed together in the universal mixer for 30 seconds, resulting in a biomimetic restorative material having an initial pH of 12 that was easy to apply and cured (according to ISO 6876:2012) within 5 to 60 minutes, depending on the particle size. During this process, compressive strengths of up to 51 MPa could be achieved when determined according to ISO 9917-1:2007(E). Furthermore, solubilities of less than 3% and flowabilities of 16 mm to 21 mm (depending on water addition) (ISO 6876:2012) could be achieved.

Example 6

Preparation of Biomimetic Dental Cements Based on Freeze-Dried Apatite-Gelatine Composites

(36) The procedure here was basically the same as for the wet apatite-gelatine composites, except that here the mixing and grinding of all dry components could take place before the addition of water, since the dried composite did not yet initiate cementation of the calcium salts and phosphate salts. The reaction was then only initiated by the addition of water, resulting in good shelf life of the cement mixture.

(37) A specific example thereof given in the following is the formation of the biomimetic dental cement based on freeze-dried apatite-gelatine composites:

(38) 0.7 g -tricalcium phosphate (2.26 mmol) having an average particle size in the range of 1-10 m was dry-triturated together with 0.042 g CaO (0.75 mmol) as well as 0.0252 g NaF (0.6 mmol). In parallel, a second mixture consisting of 0.04 g of freeze-dried apatite-gelatine composite (protein content 20%; apatite content 80%) together with 0.017 g of transglutaminase (e.g. ultrafiltration from Ajinomoto Activa WM) as well as 0.0085 g of casein and 0.3 ml of H.sub.2O were mixed together for 20 seconds in the universal mixer. Both phases were then mixed in the universal mixer for 30 seconds, resulting in an easy-to-apply and fast-curing biomimetic filling material.

Example 7

Preparation of Biomimetic Dental Cements Based on Wet Octacalcium Phosphate-Gelatine Composites

(39) For the preparation of a biomimetic dental cement based on wet octacalcium phosphate-gelatine composites, the inorganic content, as well as gelatine and water content of the composites were first determined. On the basis of the calcium and phosphate contents thus obtained (for OCP: Ca/PO.sub.4=1.33 to 1) in relation to gelatine and water, it was then possible to adjust the use of calcium-containing and phosphate-containing salts such that a molar calcium to phosphate ratio of 1.5 to 1 to 1 was obtained within the entire cement composition, with 1.67 to 1 being preferred in a standard experiment (in this case, the ratio of calcium compound to protein component could be varied over the entire range). Procedurally, this was performed by grinding all dry calcium salts as well as phosphate salts and possible added fluorine- or carbonate-containing salts, either before blending thereof or afterwards. The mixing of the dry ingredients was then followed by the addition of the wet composites and, if necessary to achieve the desired viscosity, water and a gelatine crosslinker. The entire mass was then thoroughly mixed once again and could then be applied as a restorative material.

(40) A specific example thereof is the formation of the biomimetic dental cement based on wet octacalcium phosphate-gelatine composites in the following:

(41) 0.7 g -tricalcium phosphate (2.26 mmol) having an average particle size in the range of 1-10 m was dry-triturated together with 0.044 g CaO (0.97 mmol) as well as 0.0252 g NaF (0.6 mmol). In parallel, a second mixture consisting of 0.25 g wet OCP gelatine composite (water content 75%; protein content 5%; octacalcium phosphate content 20%) together with 0.017 g transglutaminase (ultrafiltration from Ajinomoto Activa WM) as well as 0.0085 g casein and 0.15 ml H.sub.2O was mixed for 20 seconds in a universal mixer. Both phases were then mixed together in the universal mixer for 30 seconds, resulting in an easy-to-apply and fast-curing biomimetic filling material having an initial pH of 12.

Example 8

Production of Biomimetic Dental Cements Based on Freeze-Dried Octacalcium Phosphate-Gelatine Composites

(42) Basically, the procedure for the production of biomimetic dental cements based on freeze-dried OCP-gelatine composites was similar to that of the wet OCP-gelatine composites. Again, the proportion of OCP to the total mass of the composite was determined to determine the addition of calcium and phosphate containing salts, with which the Ca to PO.sub.4 molar ratio of 1.5 to 1.67 to 1 was obtained at the end. The difference with the wet composites was that water was added to start the reaction.

(43) A specific example thereof given in the following is the formation of the biomimetic dental cement based on freeze-dried octacalcium phosphate gelatine composites:

(44) 0.7 g -tricalcium phosphate (2.26 mmol) having an average particle size in the range of 1-10 m was dry-triturated together with 0.044 g CaO (0.97 mmol) as well as 0.0252 g NaF (0.6 mmol). In parallel, a second mixture consisting of 0.06 g of freeze-dried OCP-gelatine composite (protein content 20%; octacalcium phosphate content 80%) together with 0.017 g of transglutaminase (ultrafiltration from Ajinomoto Activa WM) as well as 0.0085 g of casein and 0.3 ml of H.sub.2O was mixed for 20 seconds in a universal mixer. Both phases were then mixed together in the universal mixer for 30 seconds, resulting in an easy-to-apply and fast-curing biomimetic filling material having an initial pH of 12.

Example 9

Production of Biomimetic Dental Cements Based on Wet Brushite-Gelatine Composites

(45) For the production of a dental cement based on wet brushite-gelatine composites, the content of calcium, phosphate, gelatine, as well as water was determined, as in the case of the previously described OCP-based cements to calculate addition of the other calcium as well as phosphate-containing salts based on this result, to finally obtain the molar ratio of calcium to phosphate of 1.5-1.667 to 1 as well as a suitable viscosity in the final cement material. Brushite had a molar ratio of Ca/PO.sub.4 of 1 to 1. The ratio adjusted to apatite by the other salts was such that, in addition to the conversion of the salts to apatite, the composite was also converted to apatite, thus achieving direct bonding of the inorganic components throughout the entire system. In parallel, the material properties could also be further improved herein by the addition of fluorine- or carbonate-containing salts. In order to also obtain a network of the organic component of the gelatine, or more generally of the protein components, the use of crosslinkers of the protein component, which was carried out in parallel during the addition of the aqueous component, was also advantageous for the formation of the most durable dental cement material possible.

(46) A specific example thereof given in the following is the formation of the biomimetic dental cement based on wet brushite-gelatine composites:

(47) 0.7 g of -tricalcium phosphate (2.26 mmol) having an average particle size ranging from 1 to 10 m was dry-triturated together with 0.052 g of CaO (0.97 mmol) as well as 0.0252 g of NaF (0.6 mmol). In parallel, a second mixture consisting of 0.25 g wet brushite-gelatine composite (water content 80%; protein content 1%; octacalcium phosphate content 19%) together with 0.017 g transglutaminase (ultrafiltration from Ajinomoto Activa WM) as well as 0.0085 g casein and 0.15 ml H.sub.2O was mixed for 20 seconds in a universal mixer. Both phases were then mixed together in the universal mixer for 30 seconds, resulting in an easy-to-apply and fast-curing biomimetic filling material having an initial pH of 12.

Example 10

Preparation of Biomimetic Dental Cements Based on Freeze-Dried Brushite-Gelatine Composites

(48) For dental cements based on freeze-dried brushite-gelatine composites, the same method was used as for the wet version. After determining the gelatine content of the dry composites, a suitable mixture of calcium and phosphate salts was selected and mixed together in the dry state to convert the brushite portion to apatite. Again, the addition of fluoride or carbonate containing salts could lead to another improvement of the cement properties, followed by the addition of water to achieve a suitable viscosity. To improve the properties of the cements, it was also advantageous herein to add a crosslinker to the dry material before adding the water.

(49) A specific example thereof given in the following is the formation of the biomimetic dental cement based on freeze-dried brushite-gelatine composites:

(50) 0.7 g -tricalcium phosphate (2.26 mmol) having an average particle size in the range of 1-10 m was dry-triturated together with 0.052 g CaO (0.97 mmol) as well as 0.0252 g NaF (0.6 mmol). In parallel, a second mixture consisting of 0.05 g of lyophilized brushite-gelatine composite (protein content 5%; brushite content 95%) together with 0.017 g of transglutaminase (ultrafiltration from Ajinomoto Activa WM) as well as 0.0085 g of casein and 0.3 ml of H.sub.2O was mixed for 20 seconds in the universal mixer. Both phases were then mixed together in the universal mixer for 30 seconds, resulting in an easy-to-apply and fast-curing biomimetic filling material having an initial pH of 12.

Example 11

Preparation of Biomimetic Dental Cements Based on Wet Amorphous Calcium Phosphate-Gelatine Composites

(51) As amorphous calcium phosphates are able cover a very wide range of calcium to phosphate ratios from 1.2 to 1 up to 2.2 to 1, the composition, with regard to calcium, phosphate, protein component and water, was precisely determined for each newly synthesized composite to adjust the other calcium and phosphate-containing salts used on the basis of these results used to adjust a calcium to phosphate ratio of 1.5-1.667 to 1. Due to the water content of the composites as well as additionally added water, a suitable viscosity could be adjusted. By adding the other calcium- and phosphate-containing salts and adjusting them to the ratio suitable for apatite, it was possible to initiate crystallization of the amorphous composite phase to apatite, thus achieving hardening of the cement. Addition of fluoride-containing salts could also accelerate the transformation to fluorapatite.

(52) The Formation of the Biomimetic Dental Cement Based on Wet Amorphous Calcium Phosphate-Gelatine Composites:

(53) 0.7 g -tricalcium phosphate (2.26 mmol) having an average particle size in the range of 1-10 m was dry-triturated together with 0.042 g CaO (0.75 mmol) as well as 0.0252 g NaF (0.6 mmol). In parallel, a second mixture consisting of 0.233 g wet amorphous calcium phosphate-gelatine composite (water content 80%; protein content 4%; calcium phosphate content 16%, Ca/PO.sub.4 ratio 1.67 to 1) together with 0.017 g transglutaminase (e.g. ultrafiltration from Ajinomoto Activa WM) as well as 0.0085 g casein and 0.15 ml H.sub.2O was mixed for 20 seconds in the universal mixer. Both phases were then mixed in the universal mixer for 30 seconds, resulting in an easy-to-apply and fast-curing biomimetic filling material.

Example 12

Production of Biomimetic Dental Cements Based on Freeze-Dried Amorphous Calcium Phosphate-Gelatine Composites

(54) For dental cements based on freeze-dried amorphous calcium phosphate-gelatine composites, the same method was used as for the wet version. After determining the gelatine content of the dry composites, a suitable mixture of calcium and phosphate salts was selected and mixed together in the dry state. An addition of fluoride-containing or carbonate-containing salts could also be added to this mixture, followed by the addition of water to adjust a suitable viscosity. To improve the properties of the cements, it was also advantageous here to add a gelatine crosslinker to the dry material before adding the water.

(55) A specific example thereof given in the following is the formation of the biomimetic dental cement based on freeze-dried amorphous calcium phosphate-gelatine composites:

(56) 0.7 g -tricalcium phosphate (2.26 mmol) having an average particle size in the range of 1-10 m was dry-triturated together with 0.042 g CaO (0.75 mmol) as well as 0.0252 g NaF (0.6 mmol). In parallel, a second mixture consisting of 0.04 g of freeze-dried amorphous calcium phosphate-gelatine composite (protein content 20%; calcium phosphate content 80%; Ca/PO.sub.4 ratio 1.67 to 1) together with 0.017 g of transglutaminase (e.g. ultrafiltration from Ajinomoto Activa WM) as well as 0.0085 g of casein and 0.3 ml of H.sub.2O was mixed for 20 seconds in the universal mixer. Both phases were then mixed in the universal mixer for 30 seconds, resulting in an easy-to-apply and fast-curing biomimetic filling material.

Example 13

Production of Cements Based on Calcium Phosphate-Gelatine Composites and Calcium Silicates

(57) The calcium phosphate-gelatine composites obtained were mixed with cements containing calcium silicate to cure them under conditions similar to those found in the human oral cavity. The advantage of this approach was that the wet, swollen composites were thus cured by the water consumption of the setting reaction of the cement, while the cement provided an additional curing and stabilizing component. Again, additional crosslinkers of the gelatine contained were of particular advantage for the material properties of the filling materials.

Example 14

Reaction of Wet Apatite-Gelatine Composites with Calcium Silicates

(58) Apatite-gelatine composites were cured by the use of calcium silicates to the extent that all water bound in the composites was consumed by the cement added in the setting reaction thereof, thereby curing the cement.

(59) For this purpose, wet apatite-gelatine composites (in different compositions in terms of protein and water content) were used. According to the water contained, Portland cement was added in ratios between 1 mass % and 99 mass %, so that an easily moldable and applicable mass was obtained. In addition to setting by hardening of the inorganic components, another gelatine crosslinker could be added to the cement mass, which further beneficially affected the mechanical properties.

(60) A specific example thereof given in following is the formation of the bioinspired dental cement based on wet apatite-gelatine composites in combination with calcium silicate:

(61) 0.0833 g Ca.sub.2SiO.sub.4 (0.51 mmol) having an average particle size in the range of 1-10 m was dry-triturated together with 0.1667 g Ca.sub.3SiO.sub.5 (0.73 mmol) having an average particle size of 1-10 m as well as 0.01 g NaF. In parallel, a second mixture consisting of 0.125 g wet apatite-gelatine composite (water content 75%; protein content 5%; apatite content 20%) together with 0.009 g transglutaminase (e.g. ultrafiltration from Ajinomoto Activa WM) and 0.0045 g casein and 0.12 ml H.sub.2O was mixed for 20 seconds in the universal mixer. Both phases were then mixed for 30 seconds in the universal mixer, resulting in an easy-to-apply and fast-curing biomimetic filling material with curing times between 30 minutes and 5 h, depending on the particle size. During this process, compressive strengths of up to 52 MPa could be achieved when determined according to ISO 9917-1:2007(E). Furthermore, solubilities of below 7% and flowabilities of 15 mm-27 mm (depending on water addition) (ISO 6876:2012) could be achieved.

Example 15

Reaction of Freeze-Dried Apatite-Gelatine Composites with Calcium Silicates

(62) Freeze-dried apatite-gelatine composites were mixed with Portland cement at mixing ratios of 1%-99% (W/W) and blended with water in proportions of 10-70 mass % to obtain a paste-like mass. Curing the cementitious materials obtained herein was caused by the simultaneous swelling of the apatite-gelatine composites and recrystallization or setting of the Portland cement. During this process, addition of a gelatine crosslinker was also beneficial herein to the mechanical properties (hardness) of the dental cement.

(63) A specific example thereof given in the following is the formation of the bioinspired dental cement based on freeze-dried apatite-gelatine composites in combination with calcium silicate:

(64) 0.0833 g Ca.sub.2SiO.sub.4 (0.51 mmol) having an average particle size in the range of 1-10 m was dry-triturated together with 0.1667 g Ca.sub.3SiO.sub.5 (0.73 mmol) having an average particle size of 1-10 m and 0.01 g NaF. In parallel, 0.04 g of freeze-dried apatite-gelatine composite (protein content 20%; apatite content 80%) was mixed together with 0.017 g of transglutaminase (e.g. ultrafiltration from Ajinomoto Activa WM) as well as 0.0085 g of casein and 0.3 ml of H.sub.2O for 20 seconds in a universal mixer. Both phases were then mixed in the universal mixer for 30 seconds, resulting in an easy-to-apply and fast-curing bioinspired filling material.

Example 15

Reaction of Wet Octacalcium Phosphate-Gelatine Composites with Calcium Silicates

(65) Portland cement curing of the octacalcium phosphate-gelatine composites used was based on two parallel mechanisms. In a first step, the Portland cement removed the water from the OCP-gelatine composite during setting reaction thereof, resulting in the curing of the total mass. As a second curing step, the recrystallization of the octacalcium phosphate to apatite could be achieved, since calcium hydroxide was formed during the setting reaction of the Portland cement, which provided calcium ions in high excess for the recrystallization of the OCP in an aqueous environment. Thus, the simultaneous reactions provided a high degree of bonding between the two different reactants.

(66) In this reaction, octacalcium phosphate-gelatine composites with gelatine concentrations between 1% and 50% and water contents between 1% and 99% were used and mixed with proportions of Portland cement between 1% and 99% for curing.

(67) Parallel to the inorganic setting, it was also possible herein to achieve curing of the organic component by crosslinking the gelatine.

(68) A specific example thereof given in the following is the formation of the bioinspired dental cement based on wet apatite-gelatine composites in combination with calcium silicate:

(69) 0.0833 g Ca.sub.2SiO.sub.4 (0.51 mmol) having an average particle size in the range of 1-10 m was dry-triturated together with 0.1667 g Ca.sub.3SiO.sub.5 (0.73 mmol) having an average particle size of 1-10 m as well as 0.01 g NaF. In parallel, 0.125 g wet octacalcium phosphate-gelatine composites (water content 75%; protein content 5%; OCP content 20%) were mixed together with 0.017 g transglutaminase (ultrafiltration from Ajinomoto Activa WM) as well as 0.0085 g casein and 0.12 ml H.sub.2O for 20 seconds in a universal mixer. Both phases were then mixed in the universal mixer for 30 seconds, resulting in an easy-to-apply and fast-curing bioinspired filling material.

Example 17

Freeze-Dried Octacalcium Phosphate Gelatine Composites

(70) Freeze-dried octacalcium phosphate gelatine composites with gelatine concentrations in the range of 1-50 wt % were blended with Portland cement at mixing ratios of 1-99 wt % (W/W) and mixed with water to obtain a paste-like mass. Curing of the cementitious materials obtained here proceeded by the simultaneous swelling of the gelatine components and recrystallization of the OCP gelatine composites to apatite and the parallel setting of the Portland cement. During this process, addition of a gelatine crosslinker was also beneficial to the mechanical properties of the dental cement, as a stable supporting organic network was obtained.

(71) A specific example thereof given in the following is the formation of the bioinspired dental cement based on freeze-dried OCP gelatine composites in combination with calcium silicate:

(72) 0.0833 g Ca.sub.2SiO.sub.5 (0.51 mmol) having an average particle size in the range of 1-10 m was dry-triturated together with 0.1667 g Ca.sub.3SiO.sub.5 (0.73 mmol) having an average particle size of 1-10 m and 0.01 g NaF. In parallel, 0.04 g of freeze-dried OCP-gelatine composite (protein content 20%; OCP content 80%) was mixed together with 0.017 g of transglutaminase (ultrafiltration from Ajinomoto Activa WM) as well as 0.0085 g of casein and 0.3 ml of H.sub.2O for 20 seconds in a universal mixer. Both phases were then mixed in the universal mixer for 30 seconds, resulting in an easy-to-apply and fast-curing bioinspired filling material.

Example 18

Wet Brushite Gelatine Composites

(73) Portland cement curing of the Brushite gelatine composites used was based on two parallel mechanisms. First, the Portland cement removed the water from the brushite-gelatine composite during setting reaction thereof, resulting in curing of the composite portion by drying in parallel with curing of the cement. As a second parallel curing step, recrystallization of the brushite to apatite could be achieved herein, as calcium hydroxide was formed during the setting reaction of the Portland cement, which in an aqueous environment provided calcium ions in high excess for the recrystallization of the brushite. The simultaneous reactions resulted in a high degree of bonding between the two different reactants.

(74) In this reaction, brushite-gelatine composites with gelatine concentrations between 1 mass % and 50 mass % and water content between 1 mass % and 90 mass % were used and mixed with proportions of Portland cement between 1 mass % and 99 mass % for curing.

(75) Parallel to the inorganic setting, it was also possible here to achieve curing of the organic component by crosslinking the gelatine.

(76) A specific example thereof given in the following is the formation of the bioinspired dental cement based on wet brushite-gelatine composites in combination with calcium:

(77) 0.0833 g Ca.sub.2SiO.sub.4 (0.51 mmol) having an average particle size in the range of 1-10 m was dry-triturated together with 0.1667 g Ca.sub.3SiO.sub.5 (0.73 mmol) having an average particle size of 1-10 m as well as 0.01 g NaF. In parallel, 0.125 g wet Brushite gelatine composites (water content 70%; protein content 1%; OCP content 29%) were mixed together with 0.017 g transglutaminase (ultrafiltration from Ajinomoto Activa WM) as well as 0.0085 g casein and 0.15 ml H.sub.2O for 20 seconds in a universal mixer. Both phases were subsequently mixed in the universal mixer for 30 seconds, resulting in an easy-to-apply and fast-curing bioinspired filling material.

Example 19

Freeze-Dried Brushite-Gelatine Composites

(78) Freeze-dried brushite-gelatine composites were blended with Portland cement at mixing ratios ranging from 1 mass % to 99 mass % (W/W) and mixed with water to obtain a paste-like mass. Curing of the cementitious materials obtained herein proceeded by the simultaneous swelling and recrystallization of the brushite-gelatine composites and the parallel recrystallization and setting of the Portland cement. During this process, the addition of a gelatine crosslinker was also beneficial for the properties of the dental cement.

(79) A specific example thereof given in the following is the formation of the bioinspired dental cement based on freeze-dried brushite-gelatine composites in combination with calcium silicate:

(80) 0.0833 g Ca.sub.2SiO.sub.4 (0.51 mmol) having an average particle size in the range of 1-10 m was dry-triturated together with 0.1667 g Ca.sub.3SiO.sub.5 (0.73 mmol) having an average particle size of 1-10 m and 0.01 g NaF. In parallel, 0.04 g of freeze-dried brushite-gelatine composite (protein content 5%; brushite content 95%) was mixed together with 0.017 g of transglutaminase (ultrafiltration from Ajinomoto Activa WM) as well as 0.0085 g of casein and 0.3 ml of H.sub.2O for 20 seconds in a universal mixer. Both phases were then mixed in the universal mixer for 30 seconds, resulting in an easy-to-apply and fast-curing bioinspired filling material.

(81) Further details, advantages and features of the present invention will be apparent from the following description of embodiments based on the drawing, wherein:

(82) FIG. 1 is a process schematic schematizing the process steps for the production of a medical or dental product according to one embodiment,

(83) FIG. 2 is an overview of curing achieved with different crosslinkers, and

(84) FIG. 3 is a schematic representation of a test set-up for the production of calcium phosphate-protein component composites.

(85) FIG. 1 shows a schematic diagram of the steps in a process for producing a medical or dental product according to one embodiment. Medical or dental products especially are dental substitute materials, bone substitute materials, root canal sealers, root filling materials, retrograde filling materials, pulp capping materials or perforation sealing materials.

(86) The process comprises a first step 100 of forming a composite compound of at least one calcium compound selected from the group consisting of: Calcium phosphates, calcium fluorides and calcium fluorophosphates and hydroxyl derivatives and carbonate derivatives of these calcium salts, calcium hydroxides and calcium oxides and at least one protein component selected from proteins and protein hydrolysates. Herein, the calcium compound is precipitated in the presence of the protein component.

(87) This may be followed by process step 200, wherein crosslinking of the composite compound using at least one crosslinking agent is carried out. The crosslinking agent is preferably selected from the group consisting of: transglutaminase, sortase A, tyrosinase, laccase, peroxidase, lysiloxidase, aminoxidase, glutaraldehyde and (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide/N-hydroxysuccinimide, genipin, caffeic acid, hexamethylene diisocyanate, proanthocyanidin and formaldehyde, wherein casein may be added additively during crosslinking.

(88) The material is then cured to obtain the medical product by added or contained water.

(89) Alternatively or in addition to the process step 200, setting the composite compound with uncured cement to further improve the hardness of the product to be produced, may occur as a process step 300. Preferably, the cement is selected from the group consisting of calcium silicate cement, calcium phosphate cement and mixtures thereof.

(90) FIG. 2 shows an overview of hardnesses achieved with different crosslinkers, which were determined according to Vickers HV0.3 with a Zeiss Miniload and Hardsoft measuring system. The hardness measurement was thus performed according to Vickers HV0.3 in each case, see Metallic materialsHardness testing according to VickersPart 1: Test method (ISO 6507-1:2018); German version EN ISO 6507-1:2018.

(91) The figure shows the effect of an aqueous solution of a crosslinker on the hardness of the composite material. Herein, the procedure was that two grams of a wet apatite composite (water content 75%; protein content 5%; apatite content 20%) were crosslinked with 10 ml of a crosslinker solution indicated in the diagram for 24 h. The samples were then centrifuged and dried in an oven at 50 C., cut and polished, and their hardness subsequently was determined at room temperature. The results clearly show that a mixture of transglutaminase (e.g. from Ajinomoto Activa WM after ultrafiltration through a 10000M sieve) and casein provides the best crosslinking properties and thus high hardness. In this context, the mixture of 3% transglutaminase and 1.5% casein should be particularly highlighted, since this, in combination with the apatite composite, results in a hardness of the material which is above the hardness of dentin.

(92) FIG. 3 is a schematic representation of an experimental set-up for the preparation of calcium phosphate-protein component composites. At least one protein component dissolved in water and a calcium compound are placed in a water bath 3 which is temperature-controlled by a heating device 2. Alternatively, a protein component and a phosphate compound may also be introduced. With the provision that a calcium compound has been introduced, at least one phosphate-containing compound is subsequently added. Provided that a phosphate-containing compound has been submitted, at least one calcium compound is subsequently added. In addition, the pH of the solution can be brought into a desired range and maintained by adding an acid or an alkali. A stirrer 4 is provided in the vessel 1 for stirring at the desired speed.

(93) In addition to the foregoing written description of the invention, explicit reference is hereby made to the graphic representation of the invention in FIGS. 1 to 3 for supplementary disclosure thereof.

LIST OF REFERENCE NUMBERS

(94) 1 Vessel 2 Heating device 3 Water bath 4 Stirrer 100-300 Process steps