Antimicrobial preparation

Abstract

The invention relates to an antimicrobial, antiseptic and biodegradable preparation comprising at least calcium hydroxide, polyvinylpyrrolidone (PVP) and collagen. The invention also relates to a free-flowing injectable antimicrobial, antiseptic and biodegradable paste which is produced from the preparation by adding water. The invention also relates to the use of the preparation and / or the paste as a medicament for tissue infections.

Description

[0001] patent application

[0002] Antimicrobial preparation

[0003] The present invention relates to the development of a new biomaterial that has antimicrobial properties, is biodegradable, and can be used in medicine to support healing processes, e.g., wound healing, bone healing, and during the implantation of a bone implant. The preparation according to the invention is also intended to prevent or reverse bone loss resulting from peri-implantitis (implant-associated infection) and to promote bone regeneration after implantation, thereby accelerating the integration of the implant into the bone.

[0004] Implants (endoprostheses) are artificial materials that are permanently or for extended periods implanted into a human or animal body to perform sensory, neurological, cardiovascular, orthopedic, analgesic, contraceptive, or cosmetic functions. Examples include various forms of joint replacement (e.g., hip endoprostheses), implants for osteosynthesis (e.g., plates and screws), and dental implants as anchors for artificial teeth. A potential side effect of endoprosthesis implantation is contamination of the implant or the surgical field with bacteria or other pathogens, leading to peri-implantitis or bone-implant infections. Trauma surgeons, orthopedic surgeons, and oral and maxillofacial surgeons frequently encounter such implant-associated (implant-related) bacterial infections (peri-implantitis), particularly with implants in the bone.Therefore, there is a great need for a material to prevent or treat tissue infections following the implantation of an endoprosthesis.

[0005] The number of serious bacterial infections has been increasing for years in line with the number of surgical procedures. The occurrence of peri-implantitis in a patient has serious clinical consequences that can lead to delayed healing, amputations, and even sepsis-related death. When peri-implantitis

[0006] When TM 1189 occurs, the infected implant often has to be surgically removed. This not only places an additional burden on the affected patient but also increases healthcare costs.

[0007] A serious consequence of peri-implantitis is bone loss due to infection. This bone loss has serious repercussions for the affected patient and is therefore a particular focus of implant surgery.

[0008] State of the art

[0009] To prevent peri-implantitis or treat its consequences (such as bone loss), the type of implant and its material properties must be considered. For example, bioactive coatings, surface modifications, or nano-silver technology can be used to modify implants in a way that reduces the risk of peri-implantitis. However, these coatings and modifications increase the cost of the implants, and there is limited clinical data on their effectiveness.

[0010] The use of local antibiotics in the prevention and treatment of peri-implantitis is considered effective and has the advantage over systemic antibiotic therapy of being less expensive and achieving significantly higher antibiotic concentrations at the targeted sites. However, the infected implant usually needs to be surgically removed before local antibiotic application. Most current treatment methods then require a second procedure to remove the antibiotic carrier, resulting in a total of four additional surgical interventions (implant removal, local treatment, carrier removal, and finally, implant replacement). Furthermore, known antibiotic allergies further restrict antibiotic use. In addition, bacterial resistance to certain antibiotics is increasingly common.

[0011] Although surgical therapy for peri-implantitis, namely implant removal and re-implantation after the infection has healed, is considered the most effective

[0012] While the TM 1189 treatment option is described, the success rate is only around 50%. Patients must bear the resulting costs themselves, or these costs are covered by health insurance. For example, the average cost for a single dental implant is approximately €2,500 in the case of peri-implantitis and surgical implant removal. Furthermore, the patient must wait 6-12 months for the peri-implantitis to heal before a new dental implant can be placed. Afterward, another 4-6 month waiting period is required before the prosthetic part of the dental prosthesis can be fitted. The total costs here amount to over €5,000 for the lost and the new dental implant. Additionally, the patient must undergo several surgeries, and the waiting time until the dental implant can be placed is significantly extended.

[0013] Another approach is the use of medical devices designed to prevent peri-implantitis through antisepsis. Some of these non-antibiotic medical devices are commercially available, such as calcium hydroxide and iodoform (iodoform according to LUPAC nomenclature). Iodoform can also be supplied as a gel. However, they are mostly used for cleaning defect areas, without reliable clinical data on their long-term efficacy against peri-implantitis. Calcium hydroxide is inherently highly alkaline and can therefore kill bacteria. It has anti-inflammatory and even antimicrobial properties. Due to these properties, calcium hydroxide is used in dentistry for root canal treatment. The success rate for treating inflammation during root canal treatment is reported to be 96%.Accordingly, calcium hydroxide is used clinically to treat periapical lesions and promote healing. However, calcium hydroxide has the following disadvantages when used as an antimicrobial preparation:

[0014] - Decomposition in the presence of iodoform: Calcium hydroxide can decompose iodoform in a strongly alkaline environment, reducing its bactericidal effect. The release of highly reactive intermediates such as diiodocarbene leads to reduced stability of the preparation.

[0015] TM 1189 - Toxic properties: Calcium hydroxide has strong basic properties which, at higher concentrations, can lead to tissue damage and toxic reactions, impairing healing and causing undesirable side effects.

[0016] - Lack of biodegradability: Calcium hydroxide is not fully biodegradable, which means it can remain in the body and cause long-term reactions or complications.

[0017] - Insufficient support for bone regeneration: When used alone, calcium hydroxide does not provide sufficient support for bone regeneration. The necessary biological processes for promoting healing and implant integration are lacking.

[0018] - Insufficient viscosity and flow properties: Calcium hydroxide does not possess the desired viscosity and flow properties for uniform distribution, which can lead to rapid hardening or uneven distribution. Therefore, the effectiveness of a preparation containing calcium hydroxide is unreliable.

[0019] Iodoform gradually degrades upon contact with tissue, releasing free iodine into the tissue fluid. This gives it a strong disinfectant effect, similar to iodine / potassium iodide solution, which directly contains free iodine. It is equally effective in preventing infections and promoting healing. Therefore, this antiseptic is used in dental practice for cystic defects and to heal soft tissue. The iodine has an analgesic effect and is readily absorbed. However, iodoform has the following disadvantages when used as an antimicrobial preparation:

[0020] - Uncontrolled release of iodine: Iodoform gradually degrades upon contact with tissue, releasing free iodine into the tissue fluid. This can lead to a strong disinfectant effect, but also to an uncontrolled and potentially harmful release of iodine, which may compromise the safety and efficacy of the application.

[0021] - Lack of long-term data: Reliable clinical data on the long-term effects of iodoform against peri-implantitis are lacking. Without this data, it

[0022] TM 1189 makes it difficult to assess the long-term efficacy and safety of iodoform as a monotherapy.

[0023] - Limited antibacterial effect: While iodoform has a disinfectant effect, it is not as effective as other combination antimicrobial preparations. Its use alone is therefore insufficient to ensure the complete elimination of bacteria necessary for the successful treatment of infections.

[0024] - No bone-regenerating effect: Iodoform has no bone-building effect. Therefore, it is not suitable on its own to promote bone formation after implant placement or to prevent or reverse bone loss resulting from peri-implantitis.

[0025] - Toxic properties: Iodoform can be toxic in higher concentrations and cause tissue damage. This can impair healing and lead to undesirable side effects, especially with long-term use.

[0026] Collagen is a protein that, as a structural protein, arranges itself into fibrous bundles and forms the basis of connective tissue and the extracellular matrix in animals and humans. Various types of collagen are known. Collagen is found, among other places, in the white, inelastic fibers of tendons, ligaments, bones, cartilage, and also in the subcutaneous tissue. Collagen can be enzymatically degraded in the bodies of animals and humans, for example, by collagenases from fibroblasts. Collagen is used, for example, in the chemical and pharmaceutical industries for the coatings of tablets (hard and soft capsules) and for gelatin suppositories. Collagen is also used in gelatin form for hemostatic sponges and as a blood plasma substitute. However, collagen has the following disadvantages when used as an antimicrobial preparation:

[0027] - Lack of antimicrobial effect: Collagen does not have sufficient antimicrobial activity to prevent or treat tissue infections. This means that it is not effective on its own against bacterial infections that can occur during implant procedures.

[0028] TM 1189 - Lack of long-term stability: Collagen can be enzymatically broken down in the body, which can impair its long-term stability and effectiveness. This property makes it unsuitable for applications requiring a continuous presence in tissue.

[0029] - Limited mechanical properties: Collagen alone does not possess the necessary mechanical properties to serve as a stable support material in medical applications. It can quickly lose its shape or degrade under stress.

[0030] - Lack of bone-regenerating effect: Collagen alone does not have a sufficient ability to promote bone regeneration. It must be combined with other substances that support this property in order to be effective in healing and regenerating bones.

[0031] Polyvinylpyrrolidone (PVP), also known as povidone or polyvidone, is a linear polymer of the compound vinylpyrrolidone. Vinylpyrrolidone has the molecular formula CeHgNO₃. The CAS number for polyvinylpyrrolidone is 9003-39-8. PVP is a hygroscopic, amorphous powder with a white to pale yellow color. The molar masses of commercially available polymers range from approximately 2,500 to 2,500,000 Daltons. Polyvinylpyrrolidone is used as an excipient in the pharmaceutical industry, for example, as a binder for tablets, in eye drops, or as a plasma expander. However, polyvinylpyrrolidone (PVP) has the following disadvantages when used as an antimicrobial preparation:

[0032] - Lack of antimicrobial activity: PVP alone does not have sufficient antimicrobial activity to prevent or treat infections. It offers no protection against bacterial infections, which limits its use on its own in medical preparations.

[0033] - No bone-regenerating effect: PVP alone has no ability to promote bone regeneration. It does not support the formation and healing of bone tissue, which is necessary in the treatment of bone diseases or implant procedures.

[0034] - Limited mechanical properties: PVP alone does not offer the necessary mechanical properties to be used as a stable substrate material in

[0035] TM 1189 is intended for medical applications. It may not possess the desired strength and stability.

[0036] - Lack of biodegradability: Although PVP is considered biocompatible in some applications, it is not fully biodegradable, which can lead to long-term residues in the body. This can cause potential complications and negative long-term effects.

[0037] A mixture of polyvinylpyrrolidone with 3% iodine is used in wound care in the form of a solution, ointment, or cream, for example under the trade names Betaisodona® or Braunol®. However, these preparations do not have a bone-building effect and also exhibit the disadvantages of polyvinylpyrrolidone.

[0038] EP 3 273 948 B1 describes a preparation with antimicrobial properties. It is administered in capsule form. However, the preparation is not biodegradable and is not intended for the treatment of bones.

[0039] EP 2 683 421 B1 describes an antibiotic-containing implant coating, which, however, is not suitable for use on a patient's bone. Furthermore, a surface coating is not a pharmaceutical preparation.

[0040] EP 1 244 434 B1 discloses chemotherapeutic agents for the treatment of dental wounds and thus also provides a preparation with antimicrobial properties. Although it can be applied in gel form, it is not biodegradable and is not intended for the treatment of bone.

[0041] Tasks

[0042] The object of this invention is to provide an injectable, antiseptic, antimicrobial, and biodegradable preparation that can be used medically for the prevention or treatment of peri-implantitis (implant-associated infections), particularly in the bone area. It is also intended to

[0043] TM 1189 The preparation according to the invention is intended to prevent or reverse bone loss resulting from peri-implantitis. Likewise, the preparation according to the invention is intended to promote bone formation after the implantation of an implant and thus accelerate the osseointegration of the implant, i.e., stimulate bone regeneration. Overall, the invention is intended to reduce both the frequency and severity of peri-implantitis, reduce or reverse its consequences, and improve the osseointegration of an implant.

[0044] Solution to the tasks

[0045] These problems are solved according to the invention by a preparation according to the claims. The preparation comprises calcium hydroxide, polyvinylpyrrolidone (PVP), and collagen. The preparation according to the invention comprises the following w / w percentages: 30.0–40.0% calcium hydroxide, 17.0–22.5% polyvinylpyrrolidone, and 27.0–37.5% collagen. In a first embodiment, the preparation according to the invention is in powder form. In one embodiment, the preparation according to the invention can be processed with water to form a flowable paste. In another embodiment, the water used is distilled water. In a further embodiment, the water used is sterile water. In a further preferred embodiment, the water used is distilled sterile water.Taking into account the added water, the weight percentages of the inventive paste produced from the preparation according to the invention are 30.0-34.0% w / w calcium hydroxide, 17.0-19.0% w / w polyvinylpyrrolidone, 27.0-31.0% w / w collagen and 16.5-18.0% w / w water.

[0046] The combination of calcium hydroxide and collagen in the preparation ensures effective killing and inhibition of microorganism growth. While calcium hydroxide destroys the microorganisms through its alkaline effect, collagen supports the antimicrobial effect through physical inhibition and potentially through specific antimicrobial peptides. Polyvinylpyrrolidone

[0047] TM 1189 contributes to the stabilization and better distribution of the active ingredients, which further improves the effectiveness of the preparation.

[0048] The preparation according to the invention and the paste according to the invention produced from the preparation according to the invention are biodegradable. The biodegradability is mainly due to collagen. Polyvinylpyrrolidone (PVP) also contributes to the biodegradability.

[0049] The preparation according to the invention is antiseptic and antimicrobial. The paste produced from the preparation according to the invention is injectable, antiseptic, and antimicrobial.

[0050] The preparation according to the invention and the paste according to the invention produced from the preparation according to the invention are suitable for the prevention or treatment of peri-implantitis (implant-associated infections), especially in the bone area.

[0051] The preparation according to the invention and the paste according to the invention produced from the preparation according to the invention prevent bone loss as a result of peri-implantitis or reverse bone loss as a result of peri-implantitis.

[0052] The preparation according to the invention and the paste according to the invention produced from the preparation according to the invention promote bone formation after the implantation of an implant and thus accelerate the healing of the implant into the bone, thereby stimulating the bone regeneration capacity.

[0053] The preparation according to the invention and the paste produced from it reduce both the frequency and severity of peri-implantitis, reduce or reverse its consequences, and improve the osseointegration of an implant. The preparation and the paste produced from it promote good bone regeneration. Bone regeneration is understood to mean the growth of new bone cells and the formation of new bone tissue. Bone regeneration depends on many factors, which are known to those skilled in the art, and whose qualitative assessment is also known to them.

[0054] TM 1189 The combination of collagen and calcium hydroxide in the preparation according to the invention ensures bone regeneration capacity. Collagen provides structural support for the growth of new bone cells and tissue, while calcium hydroxide promotes mineralization and the formation of new bone tissue by providing calcium ions and creating a conducive environment for bone healing.

[0055] The preparation according to the invention and the paste according to the invention produced from the preparation according to the invention are suitable for prophylactic or therapeutic use in humans or vertebrates. They thus fulfill all the tasks assigned to them.

[0056] Through our own scientific research, we have succeeded in developing a preparation that reduces the toxic properties of calcium hydroxide. This was achieved by combining it with collagen, resulting in a high antimicrobial effect, combined with excellent biodegradability and stimulation of bone regeneration.

[0057] It has been found that collagen, calcium hydroxide, and polyvinylpyrrolidone can be mixed with water to form a flowable paste exhibiting various advantageous medicinal properties. Without being bound to a specific theory, it can be assumed that this particular combination of calcium hydroxide with collagen and polyvinylpyrrolidone leads to an unexpected stabilization of the individual components, which could not have been anticipated. This stabilization can presumably be attributed to the effect of collagen and polyvinylpyrrolidone in combination as a kind of "protective colloid." As a result, the preparation according to the invention has an almost unlimited shelf life. The paste produced from the preparation according to the invention has a shelf life of at least three weeks.

[0058] While the effect of polymeric substances as protective colloids is already known in other areas, it could not have been expected a priori here, since the strongly basic effect of calcium hydroxide was expected to have a decomposing effect on collagen as a protein and thus lead to its rapid destruction. The binding of the water content in the paste according to the invention through the formation of a protective colloid has the technical effect that the

[0059] TM 1189 calcium hydroxide is stabilized because the water molecules bound in the protective colloid are no longer available for the reaction of the carbon dioxide from the environment with the calcium hydroxide, thus delaying its reaction to calcium carbonate.

[0060] The unexpected stability and effectiveness of the paste produced from the preparation according to the invention can be attributed - at least partially - to its pasty character, since the collagen and the polyvinylpyrrolidone bind the water molecules contained in the pasty preparation by forming hydrogen bonds.

[0061] Production of the inventive paste produced from the preparation according to the invention:

[0062] The preparation according to the invention consists of a mixture of calcium hydroxide, polyvinylpyrrolidone (PVP), and collagen. A flowable paste is produced from this preparation by mixing it with water, for example, distilled and / or sterile water. In a preferred embodiment, a paste with a ratio of 40.0% w / w calcium hydroxide, 22.5% w / w polyvinylpyrrolidone, and 37.5% w / w collagen was prepared. To produce the paste, the components calcium hydroxide, polyvinylpyrrolidone powder, and collagen powder were weighed in the specified proportions and mixed with a sufficient quantity of water to form a paste.

[0063] For the preparation according to the invention, polyvinylpyrrolidone PVP10, i.e., a polyvinylpyrrolidone with a molecular mass of approximately 10,000 Daltons, has proven to be particularly suitable. The tests carried out showed that the use of PVP10 positively influences the flow properties, setting behavior, and shelf life of the preparation. In addition, PVP10 has the following advantages over other PVPs:

[0064] - Lower molecular weight: PVP10 has a lower molecular weight (approx. 10,000 Daltons) compared to other PVP variants such as PVP40. This results in better flow properties and a lower viscosity of the paste, which facilitates handling and application. - Better solubility: PVP10 is more soluble in water than variants with higher molecular weights. This leads to a more even distribution in the paste and ensures a consistent and homogeneous mixture of the ingredients.

[0065] - Improved stability: PVP10 contributes to the stabilization of the other components of the paste, especially when combined with collagen and calcium hydroxide. This stability is important to ensure the paste's shelf life and effectiveness over extended periods.

[0066] - Reduced viscosity: The lower viscosity of PVP10 allows for easier injection of the paste, which is particularly important for medical applications where precise and controlled application is required.

[0067] - Biocompatibility: PVP10 is biocompatible and is used in many pharmaceutical and medical applications because it is well tolerated and does not cause any unwanted reactions in the body.

[0068] - Moisture binding: PVP10 has excellent moisture-binding properties, which improves the hydration and stability of the paste and helps to maintain the desired physical properties of the paste.

[0069] The molecular weight of the polyvinylpyrrolidone used directly influences the flow properties (viscosity, flow resistance) of the paste-like preparation. If different flow properties are desired, or if a different type of collagen is to be used, a different type of polyvinylpyrrolidone (such as PVP40) can be employed to achieve the desired flow properties. Experts are aware that the flow properties of paste-like preparations depend on the properties and proportions of the starting materials, so there are always several possible variations regarding the types and proportions of the starting materials for adjusting the flow properties. The combined use of different PVPs, such as PVP10 and PVP40, is also possible. The following preparations are possible to achieve specific desired properties:

[0070] - Combination of PVP10 and PVP40: This combination offers the advantage of a balanced viscosity and improved mechanical properties. PVP10 ensures good solubility and easy processing, while PVP40 increases the paste's strength and stability. This combination can be used when both good injectability and increased mechanical strength of the paste are required.

[0071] - Combination of PVP10 and PVP K30: This combination has the advantage that PVP K30 has a medium molecular weight (approx. 30,000 Daltons) and can therefore be a compromise between using PVP10 and PVP40. It combines the good solubility of PVP10 with a moderate increase in viscosity and strength. This combination is useful when a paste with medium viscosity and sufficient stability is required.

[0072] - Use of PVP-VA copolymers: These copolymers of PVP and vinyl acetate (e.g., PVP / VA S630) can offer specific properties such as improved adhesion and flexibility. These copolymers are particularly advantageous in applications where good adhesion to tissue is required. They are ideal for situations where the paste needs to stay firmly in place, such as in the treatment of bone defects or for wound healing.

[0073] - Combination of PVP10 and PVP K90: PVP K90 has a high molecular weight (approx. 90,000 Daltons) and can significantly increase viscosity and mechanical strength. In combination with PVP10, controlled adjustment of the flow properties can be achieved. This combination can be used when a very firm and stable paste is required that is still sufficiently flowable for injection.

[0074] The flow properties can be further optimized by adjusting the proportions of collagen, calcium hydroxide, and PVPs. For example, a

[0075] TM 1189: A higher proportion of PVP10 improves solubility and injectability, while a higher proportion of PVP40 or PVP K90 increases strength.

[0076] The flow properties can also be adjusted by introducing additives to meet specific requirements. This can be achieved, for example, by adding plasticizing or thickening agents such as glycerin or polyethylene glycol (PEG).

[0077] Production on a laboratory scale:

[0078] All percentages mentioned below as examples are given as weight percent (% w / w), even if they refer to liquids. To produce the pasty preparation according to the invention on a laboratory scale, 3.37 g of collagen powder, 4 g of calcium hydroxide, and 2.25 g of polyvinylpyrrolidone powder are weighed out. To create a flowable but solid mass (solid in the sense of pasty, i.e., malleable and retaining its shape at least partially for at least a few seconds), each substance is mixed with just enough distilled and sterile water to produce a flowable yet malleable and shape-retaining paste. This paste remains flowable yet malleable and retains its shape for at least several hours. The paste therefore exhibits thixotropic properties because it becomes more flowable under shear stress and returns to a more solid form when at rest.This thixotropic behavior is particularly advantageous because it makes it well suited for 3D printing.

[0079] Table 1: Dry substances of the preparation according to the invention for the production of a paste as well as exemplary values ​​of the required water addition (in ml per gram of the respective substance), which - depending on the moisture content and / or particle size of the starting substances - may vary by up to 20% around the respective specified value.

[0080] The calcium hydroxide powder is spread out on a clean surface and moistened drop by drop with measured distilled sterile TM 1189 water using a disposable pipette. A spatula is used to mix the calcium hydroxide powder with the water. Water is added to the calcium hydroxide gradually until a paste-like, mixable mass is formed. Next, the collagen is mixed drop by drop with measured distilled sterile water using a spatula. Only enough water is added to form a paste. This paste should have a flowable yet malleable consistency, which can be described as "pasty" and "crumbly sticky." This consistency allows the paste to hold its shape for a few seconds before deforming under shear and, if necessary, returning to its original shape.

[0081] Polyvinylpyrrolidone powder is then mixed in small amounts with measured distilled sterile water to form a paste. All substances are then combined and mixed. Mixing continues until the collagen, calcium hydroxide, and polyvinylpyrrolidone have formed a solid but still flowable (i.e., paste-like) mass. An additional 0.1 ml of distilled sterile water is added to the mixture to ensure the paste is flowable. The paste is then filled into a suitable container, such as disposable syringes, and is ready for use.

[0082] The viscosity and flow resistance of the preparation according to the invention, i.e. the paste, can be illustrated by the following general formula:

[0083] Here, F represents the flow resistance in Newtons (N), q the viscosity in Pascal seconds (Pa * s), v the flow velocity in meters per second (m / s), and d the diameter of the flow channel in meters (m).

[0084] Following the procedure described above, further formulations of the preparation according to the invention were produced and tested on a laboratory scale. A compilation of the variants tested to date is included in Table 2, together with a general composition according to the invention extrapolated from the present examples, encompassing the concentration ranges of the components collagen, calcium hydroxide, and polyvinylpyrrolidone. In addition to the specifically mentioned components, the preparation according to the invention may contain further additives such as thickening agents, reaction retarders, stabilizers, buffer substances, and the like. These modify the properties of the preparation or the paste produced from the preparation according to the invention (for example, flowability, stability, viscosity, pH value).The specified upper and lower limits of the concentration ranges are therefore to be understood as meaning that the contents of each specific mixture according to the invention of the components collagen, calcium hydroxide and polyvinylpyrrolidone as well as distilled water each amount to 100% by weight.

[0085] The weight percent of collagen is 27-31% w / w, the weight percent of calcium hydroxide is 30-34% w / w, the weight percent of polyvinylpyrrolidone is 17-19% w / w, and the weight percent of distilled sterile water is 16.5-18% w / w.

[0086] The production of the preparation according to the invention and the paste produced from it is arbitrarily scalable, since the percentage proportions of the substances remain constant. The production of large batches requires only a corresponding adjustment of the equipment and continuous monitoring of the mixing processes to ensure the homogeneity and consistency of the preparation according to the invention and the paste produced from it. Controlling the viscosity and consistency of the preparation according to the invention is also possible.

[0087] The quality of the preparation and the paste produced from the preparation according to the invention can be ensured by regular quality controls.

[0088] The preparation according to the invention and the paste produced from it are used in surgery to prevent or treat tissue infections, particularly in the bone area. They are used prophylactically by being added to the surgical field before the implantation of an endoprosthesis. They are also used therapeutically in the early stages of an infection (within the first three weeks) by applying the preparation or the paste produced from it around the infected area to preserve the implant and prevent its loss. The preparation and the paste produced from it are also used to treat peri-implantitis in the surgical field after the removal of endoprostheses.In addition to the prophylaxis and treatment of peri-implantitis, another potential application is prophylactic use in open fractures with a high risk of infection, as a first-aid measure by paramedics, e.g., in cases of gunshot wounds or traffic accidents. The application is diverse and encompasses all medical treatments related to peri-implantitis in bones or injuries with exposed bone (open fractures).

[0089] Examples of implementation

[0090] Inhibitory effect of the individual components of the preparation according to the invention and of the paste according to the invention produced from the preparation according to the invention on E. coli bacteria

[0091] Experiment 1: Preventing bacterial growth

[0092] To verify whether the inventive paste produced from the preparation according to the invention prevents the growth of E. coli bacteria, the

[0093] TM 1189 performed the following experiment. A Petri dish containing agar-agar medium was removed from the refrigerator, and a swab of previously cultured E. coli bacteria was diluted with 1 ml of distilled water. This swab was spread onto the medium using a Drigalski spatula. The Petri dish was sealed, and the bacteria were allowed to dry for up to 15 minutes.

[0094] In order to compare the individual effects of the individual components of the preparation according to the invention with the effect of the paste according to the invention produced from the preparation according to the invention, the individual components were mixed with water to form comparison pastes (calcium hydroxide comparison paste, polyvinylpyrrolidone comparison paste, collagen comparison paste).

[0095] Using a template and a sterile pipette tip, test holes were pierced into the nutrient medium to evenly distribute the comparison pastes and the inventive paste produced from the preparation according to the invention in flowable form and in the hardened state (see Table 3).

[0096] Table 3: Components of the inventive paste produced from the preparation according to the invention, individually and in combination according to the invention, as well as distilled sterile water as a reference substance (negative control) for the prepared wells in the agar-agar medium. The wells were filled as follows: 150 pl of calcium hydroxide reference paste was pipetted into the first well using a pipette. The second well was filled with 150 pl of polyvinylpyrrolidone reference paste, and 150 pl of collagen reference paste was added to the third well. 0.1 g of the hardened inventive paste produced from the preparation according to the invention was filled into the fourth well, corresponding to 150 pl of the still-flowable inventive paste produced from the preparation according to the invention before hardening.Into the fifth well of the agar-agar medium, 150 µl of the flowable paste prepared according to the invention was added. Into the sixth well, 150 µl of distilled sterile water was pipetted as a control. The Petri dish was sealed and placed in the incubator for 8 hours at the optimal growth temperature for E. coli bacteria. After this incubation period, the results were documented to determine whether bacterial growth had been inhibited (see Figure 1).

[0097] Figure 2 shows that in test 1, the radial extent of the inhibition zones in the hardened paste produced from the inventive preparation reached an average value of over 1.33 cm. In the flowable paste produced from the inventive preparation, the average radial extent of the inhibition zones was 0.66 cm. The collagen comparison paste had an average inhibition zone diameter of approximately 0.38 cm, while the polyvinylpyrrolidone comparison paste showed the lowest value with an average of only 0.26 cm.

[0098] Attempt 2: Killing the bacteria

[0099] To verify whether the inventive paste produced from the preparation according to the invention kills any existing E. coli bacteria, a second experiment was conducted. A hardened agar-agar medium was removed from the refrigerator. A swab of the cultured E. coli bacteria was diluted with 1 ml of distilled water and spread onto the medium. The Petri dish was sealed and incubated for 8 hours at 37°C to promote bacterial growth. After this incubation period, holes were punctured in the medium using a sterile pipette tip and the same template as in Experiment 1. The holes were filled with the same test substances as in the first experiment. The Petri dish was then sealed again and placed in the incubator for 24 hours. After this second incubation period, the Petri dish was removed and the results were documented to determine whether the bacteria had been killed.

[0100] In experiment 2, the radial extents of the inhibition zones in the hardened paste produced from the inventive preparation were smaller and had an average value of approximately 0.42 cm. The flowable paste produced from the inventive preparation showed inhibition zone diameters averaging 0.33 cm. In the second experiment, the collagen comparison paste had an average inhibition zone diameter of approximately 0.24 cm, while the polyvinylpyrrolidone comparison paste exhibited an average inhibition zone diameter of only 0.21 cm.

[0101] Experiment 3: Influence of the inventive paste produced from the preparation according to the invention on cells of mammals

[0102] In experiment 3, the effect of the inventive paste prepared from the inventive preparation on rat mesenchymal stem cells was investigated. This demonstrates the compatibility of the inventive preparation and the inventive paste prepared from the inventive preparation with living cells. Initially, the cells were counted using a CASY® cell counter. The cells were then transferred to a 6-well culture plate, and the test substances were applied to a sieve over the cells, but in contact with the medium. A control sample with an empty cell sieve was also prepared (Figure 3). The cell culture plate was sealed and incubated at 37°C. After 24 hours, the cells were examined.

[0103] The initial cell count was approximately 101,000, with 88,600 live and 12,400 dead cells. After 24 hours, the control samples showed an average of 468,313 cells / mL, of which 411,275 were live and 57,038 were dead. The flowable solution from the

[0104] The inventive paste prepared from the preparation TM 1189 resulted in significant cell growth to 1.3 million cells / mL, of which 937,500 were viable. The hardened inventive paste prepared from the inventive preparation showed lower growth with 754,800 cells / mL, of which 665,500 were viable. These results demonstrate that the flowable inventive paste prepared from the inventive preparation promotes stem cell growth, from which the bone-regenerating effect of the inventive paste prepared from the inventive preparation can be derived.

[0105] The calcium hydroxide control paste yielded 482,000 cells, of which 442,000 were viable. The polyvinylpyrrolidone and collagen control pastes showed less growth than the control. The polyvinylpyrrolidone control paste had 365,000 cells (329,000 viable), and the collagen control paste had 252,000 cells (218,000 viable). The percentage growth was calculated using the following formula:

[0106] Percentage growth = (Number of living cells after 24 hours / Total number of cells at time 0) x 100

[0107] The paste produced from the preparation according to the invention showed the highest percentage growth of living cells. These data are shown as a bar chart in Figure 4. Overall, the paste produced from the preparation according to the invention showed the greatest improvement in cell growth compared to the other components and the control.

[0108] Experiment 4: Viscosity test of the inventive paste produced from the preparation according to the invention

[0109] To investigate how the flowability of the inventive paste produced from the preparation according to the invention changes depending on the storage time, the viscosity of the inventive paste produced from the preparation according to the invention was checked over time. For this purpose, the inventive paste produced from the preparation according to the invention was

[0110] TM 1189 paste according to the invention was mixed at various times: after one day (D1), as well as after one week (1W), two weeks (2W), and three weeks (3W). These flowable pastes produced from the preparation according to the invention were prepared three, two, and one week, respectively, and one day before the viscosity test. This allowed the investigations into changes in flowability for the flowable pastes produced from the preparation according to the invention at different ages to be carried out on the same day. Two ml of the flowable pastes produced from the preparation according to the invention were filled into 5 ml disposable syringes, sealed airtight (e.g., with Parafilm®), and stored in the dark at approximately 20°C (room temperature) to prevent them from hardening prematurely. For the viscosity test, eight samples were measured for each storage period.

[0111] The viscosity of the free-flowing pastes produced from the inventive preparation as a function of storage time was investigated using a material testing machine (e.g., from ZwickRoell) and testing software (e.g., testXpert® III). The investigation was carried out on the material testing machine. Initially, all syringes were detached from the Parafilm® and each was clamped into the testing machine. The test speed was 5 mm / min. The disposable syringe was fixed vertically and upside down in a holder in the testing machine, so that the machine's plunger exerted an axial force on the piston of the disposable syringe, ejecting the free-flowing paste produced from the inventive preparation.The results, showing the force (N) with which the syringe plunger is pressed down to expel the flowable paste produced from the preparation according to the invention, were documented.

[0112] The results show how the viscosity of the inventive paste produced from the preparation according to the invention changes depending on the storage time.

[0113] The force (N) required to displace the flowable paste produced from the preparation according to the invention increased considerably from D1 to 3W. Group D1 showed the lowest force values, which suggests

[0114] TM 1189 indicates that the flowable paste produced from the preparation according to the invention was easiest to displace after one day. With increasing curing time, the force values ​​increased, with group 3W exhibiting the highest values. This suggests that the paste becomes significantly more resistant to displacement over time, probably due to increased cohesion and structural integrity during the curing process.

[0115] The displacement values ​​(m) also varied significantly over time. After one day (D1), the flowable paste produced from the preparation according to the invention showed the highest displacement, indicating greater deformability. With increasing curing time to one week (1W), two weeks (2W), and three weeks (3W), the displacement values ​​decreased, with the lowest displacement observed in the 3W group. This decrease in displacement over time correlates with the increase in force, further supporting the conclusion that the flowable paste produced from the preparation according to the invention becomes stiffer and less deformable the longer it cures.

[0116] The flow rate (m 3The flowable paste produced from the preparation according to the invention showed a decreasing trend from D1 to 3W. Group D1 had the highest flow rate, indicating that the flowable paste produced from the preparation according to the invention was most fluid after one day. With progressive curing over weeks, the flow rate decreased, with group 3W exhibiting the lowest flow rate. This indicates a significant increase in viscosity and a decrease in flowability as the flowable paste produced from the preparation according to the invention cures.

[0117] The shear rate (s -1The shear rate and shear stress (Pa) both increased with the curing time. Group D1 had the lowest shear rate and shear stress, while group 3W had the highest values. The increase in shear rate and shear stress over time shows that the resistance of the flowable paste produced from the preparation according to the invention to shear forces increases as it cures, which is due to increased internal friction.

[0118] TM 1189 and structural stability are indicated. These changes are consistent with the observed increases in force and decreases in displacement and flow rate.

[0119] The viscosity values ​​(Pa s) increased considerably from D1 to 3W, with group D1 exhibiting the lowest viscosity and group 3W the highest. This increase in viscosity over time indicates that the flowable paste produced from the preparation according to the invention becomes thicker and more resistant to flow as it cures. The higher viscosity in the later weeks is consistent with the lower flow rates and higher forces required for displacement, supporting the trend of increasing material stiffness and decreased flowability.

[0120] The results show significant differences in the flowability properties of the flowable paste produced from the inventive preparation when tested after different time periods (one day (D1), one week (1W), two weeks (2W), and three weeks (3W)). Each variable showed clear trends and differences across these time points, providing valuable insights into the temporal changes in the material properties.

[0121] The force required at constant speed to displace the flowable paste produced from the preparation according to the invention increased considerably from D1 to 3W. This indicates that the flowable paste produced from the preparation according to the invention becomes significantly more resistant to displacement over time, probably due to increased cohesion and structural integrity during storage.

[0122] The displacement values ​​also varied significantly over time. After one day (D1), the flowable paste produced from the preparation according to the invention showed the highest displacement, indicating higher deformability. With increasing storage time to one week (1W), two weeks (2W), and three weeks (3W), the displacement values ​​decreased, with the lowest displacement observed in the 3W group. This decrease in displacement over time correlates with the increase in force, which further

[0123] TM 1189 supports the finding that the flowable paste produced from the preparation according to the invention becomes stiffer and less deformable the longer it is stored.

[0124] The flow rate of the flowable paste produced from the preparation according to the invention showed a decreasing trend from D1 to 3W. This indicates a significant increase in viscosity and a decrease in flowability, as the flowable paste produced from the preparation according to the invention hardens during storage and it becomes more difficult for the material to flow.

[0125] The shear rate and shear stress both increased with storage time. The increase in shear rate and shear stress over time indicates that the resistance of the flowable paste produced from the inventive preparation to shear forces increases during storage, suggesting increased internal friction and structural stability. These changes are consistent with the observed increases in force and decreases in displacement and flow rate.

[0126] The viscosity values ​​increased considerably from D1 to 3W, indicating that the flowable paste produced from the preparation according to the invention becomes thicker and more resistant to flow during storage. The higher viscosity in the later weeks corresponds to the lower flow rates and the higher forces required for displacement, supporting the trend of increasing material stiffness and decreased flowability.

[0127] The tests show that the flowability properties of the free-flowing paste produced from the inventive preparation change considerably over time. After one day, the free-flowing paste produced from the inventive preparation is relatively easy to displace, highly deformable, and exhibits high flow rates as well as low viscosity, shear rate, and shear stress. With progressive storage over one week, two weeks, and three weeks, the free-flowing paste produced from the inventive preparation becomes significantly more resistant to displacement, less deformable, with lower flow rates, and higher viscosity, shear rate, and shear stress.

[0128] TM 1189 Viscosities, shear rates and shear stresses. These findings show that the flowable paste produced from the preparation according to the invention undergoes significant changes in its physical properties during the storage process.

[0129] The preparation according to the invention and the paste produced from the preparation according to the invention can be used prophylactically to prevent tissue infections, particularly in bones. This applies both to its use in preventing peri-implantitis during the implantation of an endoprosthesis and to other injuries of bones or other tissues, such as open fractures, gunshot wounds, or other injuries.

[0130] The preparation according to the invention can be used therapeutically to treat tissue infections, particularly in bones. This applies both to the treatment of peri-implantitis after the implantation of an endoprosthesis and to infections of bones or other tissues following injuries such as open fractures, gunshot wounds, or other injuries.

[0131] Figure caption

[0132] Figure 1 shows the agar Petri dish with bacteria and holes containing the components of the preparation according to the invention or the paste according to the invention produced from the preparation according to the invention: Hole 1 = calcium hydroxide comparison paste, Hole 2 = polyvinylpyrrolidone comparison paste, Hole 3 = collagen comparison paste, Hole 4 = hardened paste according to the invention produced from the preparation according to the invention, Hole 5 = flowable paste according to the invention produced from the preparation according to the invention, Hole 6 = distilled sterile water.

[0133] Figure 2 shows the effect of the inventive paste produced from the preparation according to the invention and the comparison pastes on E. co / / bacteria.

[0134] TM 1189 Shown is the radial extent of the inhibition zones (inhibition zone diameter) in cm.

[0135] Figure 3 shows the effect of the inventive paste and the comparison pastes produced from the preparation according to the invention on mesenchymal stem cells of rats.

[0136] Figure 4 shows the number of cells after incubation with the comparison pastes and the inventive paste (flowable and hardened) produced from the preparation according to the invention.

[0137] Figure 5 shows the percentage growth of the total number of cells to the number of living cells after incubation with the comparison pastes and the inventive paste (flowable and hardened) produced from the preparation according to the invention.

[0138] Figure 6 shows the disposable syringe containing the flowable paste produced from the preparation according to the invention, inserted into a material testing machine.

[0139] TM 1189

Claims

Claims 1. Antimicrobial preparation comprising at least calcium hydroxide, polyvinylpyrrolidone (PVP) and collagen.

2. Antimicrobial preparation according to claim 1, characterized in that it comprises 30-40% w / w calcium hydroxide, 17-22.5% w / w polyvinylpyrrolidone (PVP) and 27-37.5% w / w collagen.

3. Antimicrobial preparation according to claims 1 and 2, characterized in that it comprises 40.0% w / w calcium hydroxide, 22.5% w / w polyvinylpyrrolidone (PVP) and 37.5% w / w collagen.

4. Antimicrobial preparation according to one of claims 1 to 3, characterized in that the polyvinylpyrrolidone (PVP) used is a polyvinylpyrrolidone with a molecular weight of 10,000 Daltons (PVP10).

5. Antimicrobial preparation according to claim 4, characterized in that at least one further polyvinylpyrrolidone selected from the group consisting of PVP40 (with a molecular weight of 40,000 Daltons), PVP K90 (with a molecular weight of 90,000 Daltons) or PVP K30 (with a molecular weight of 30,000 Daltons) is used in addition to PVP10.

6. A method for producing a paste comprising the steps i. providing an antimicrobial preparation according to any one of claims 1 to 5 ii. providing water iii. mixing the preparation from step i. with the water from step ii., such that a flowable paste is formed. TM 1189 7. Method for producing a paste according to claim 6, characterized in that the water from step ii. is distilled and / or sterile water.

8. Method for producing a paste according to claims 6 and 7, wherein the weight percent of collagen is 27-31% w / w, the weight percent of calcium hydroxide is 30-34% w / w, the weight percent of polyvinylpyrrolidone (PVP) is 17-19% w / w and the weight percent of water is 16.5-18% w / w.

9. Method for producing a paste according to claim 6, characterized in that, in addition to steps i. to iii., glycerin and / or polyethylene glycol (PEG) are added in a further step iv. to adjust the flow properties of the paste.

10. Method for producing a paste according to claims 6 to 9, characterized in that, in addition to steps i. to iv., additives such as thickening agents, reaction retarders, buffer substances and / or stabilizers are added in a further step v. to modify the properties of the paste.

11. Use of the antimicrobial preparation according to any one of claims 1 to 5 and / or the paste produced according to any one of claims 6 to 10, for the manufacture of a pharmaceutical preparation.

12. Use of the pharmaceutical preparation of claim 11 as a medicament for the prophylaxis of tissue infections.

13. Use of the pharmaceutical preparation of claim 11 as a drug for the treatment of tissue infections. TM 1189 14. Use of the pharmaceutical preparation of claim 11 as a drug against tissue infections in the bone area. TM 1189

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