Patent Application
20240000548
The present invention relates to a dental implant being at least partially covered with a protective layer.
Dental implants are used to replace individual teeth or for anchoring more complex structures, which generally replace several or even all teeth. Implants have two essential parts: an anchoring part and an abutment part. The anchoring part is embedded in the bone, where it osseointegrates with the bone tissue to provide a firm anchor for the prosthesis. The abutment extends into the oral cavity and provides a support for the prosthesis. The desired prosthetic element (e.g. bridge or crown) is fastened over the abutment such that at least part of the abutment is housed within the prosthesis and provides core support to this. The prosthetic element can be adhesively bonded, cemented, screwed or directly veneered onto the abutment.
Implants, such as dental implants, are well known in the art. They generally consist of a material, which is biocompatible and which additionally has favorable mechanical properties.
In addition, it is required that the dental implant provides good osseointegration. The term “osseointegration” designates the direct structural and functional connection between living bone and the surface of the load-bearing implant. A good osseointegration means that the implant, after reaching a primary stability by screwing it into the bone, safely ossifies within a short healing time so that a permanent bond between implant and bone is obtained.
In the beginning phase of modern implantology a minimally rough surface was the gold standard. Later, an increase to moderately rough surfaces led to faster and firmer osseointegration in several experimental studies using various animal models.
A breakthrough technology in the development of highly osseointegrative dental implants is the so-called “SLA” process which is disclosed in EP 0 388 576, involving sandblasting the implant's surface followed by acid-etching to achieve an optimal topography for the attachment of bone cells.
Based on the “SLA” technology, the so-called “SLActive” surface was developed and disclosed in WO 00/44305, which further comprises conditioning the “SLA” surface either in nitrogen or in an isotonic saline solution, thereby maintaining the high hydrophilicity of the “SLA” surface which would otherwise be lost due to reaction with the atmosphere. The packaging process is time-consuming and expensive, which is a disadvantage of this procedure.
US 2014/172028 discloses a surface treatment process of an endosseous implantable medical device by covering the surface of the medical implant with a sugar or a sugar alcohol.
WO 2018/189185 discloses a dental implant made of a ceramic material. The implant surface has at least partially a contact angle of less than 20° and the implant surface is at least partially covered with a protective layer. Said protective layer comprises a water-soluble dextran having a molecular weight of more than 15′ 000 Da.
WO 2008/098976 discloses a process for the production of implants with a hydrophilic surface by covering the surface of said implant with a salty layer.
US 2009/0132048 discloses a dental implant which is at least partly provided with a protective layer which dissolves on contact with body fluid and/or the bone, said protective layer consisting of salt. However, salty protective layers, in particular those consisting of NaCl, show a reduced long-term stability.
WO 03/030957 discloses an implant having roughened hydroxylated and hydrophilic surface and being treated in the hydroxylated state with high-energy ultraviolet radiation. One disadvantage of this solution is the additional treatment step which is supposed to be carried out by the surgeon in particular.
Campbell et al disclose the effect of a keratin hydrogel on osseointegration.
The object of the present invention was to provide a protective layer for dental implants which prevents the surface from turning to hydrophobic. In addition, the protective layer needs to be biocompatible and the product should not suffer from hydrothermal aging while being hydrophilic.
The problem is solved by a dental implant according to claim 1. Further preferred embodiments are subject of the dependent claims.
It was found that a dental implant comprising an implant surface having at least partially a contact angle of less than 20°, said implant surface being at least partially covered by a protective layer comprising a keratin hydrolyzate provides an excellent osteointegration. The protective layer has preferably a water content of less than 10% by weight. Interestingly, it is no longer necessary that the dentist removes the protective layer before implanting the dental implant. Further, the protective layer ensures hydrophilicity until the contact of the implant with the bone is established. Due to its composition, the protective layer according to the present invention is biocompatible and has a good accessibility.
The protective layer according to the present invention prevents the deposition of contaminants on the implant surface having at least partly a contact angle of less than 20°. In addition, the contact angle on the surface of the protective layer is less than 20°, that is, even the protective layer itself has an excellent wettability which allows an accelerated osseointegration even without removing the protective layer. In particular, it could be shown that the protective layer according to the present invention can withstand harsh conditions such as high humidity, low temperature or low pressure. The protective layer of the dental implant according to the present invention allows to maintain the hydrophilicity of the surface during the storage time in the air.
The standard parameter to determine the hydrophilicity of a surface is the measurement of the water contact angle (DIN 55660-2). The contact angle quantifies the wettability of the implant by water and therefore the degree of contact with the hydrophilic surroundings. The term “contact angle” as used in the context of the present invention relates to the contact angle of water on the surface, i.e. to the angle formed at the interface where water meets the surface. Thereby, the term “water” used in the context of the contact angle measurement relates to pure water, specifically ultrapure water. In particular, the contact angle measurement is carried out by the sessile drop method (e.g. by means of a device of the type EasyDrop DSA20E, Kruss GmbH) using a drop size of 0.1 μL for hydrophilic and 0.3 μL for hydrophobic discs of 5 mm diameter, meaning that keratin coated discs are measured with 0.1 uL droplet. Contact angles were calculated by fitting a circular segment function to the contour of the droplet placed on the surface. The terms “hydrophilic” or “hydrophilicity” as used in the context of the present invention refer to a water contact angle of the hydrophilic surface area directly on the dental implant being less than 20°, more preferably less than 10°.
Such a hydrophilic surface having a contact angle of less than 20° can be obtained for example by the so-called SLA technology or by other etching procedures.
The expression “keratin hydrolyzate” stands for a hydrolyzate obtained from keratin wherein the peptide chains were divided into smaller peptides with lower molecular weights, that is a molecular weight of less than 20'000 g/mol. The molecular weight of the keratin hydrolysate may be determined for example by SDS-PAGE or by measuring absorption at 280 nm. A keratin hydrolyzate can be prepared, for example, by an acid or alkaline hydrolysis, or by enzymatic digestion. The keratin source usually used may be from several origins such as, for example, human hair fibers, wool, animal hair, feathers and horns.
As the keratin hydrolyzate exists in solution as colloids, it is capable of interacting with the blood and thus facilitates the protein absorption and the formation of a layer of blood components on the implant surface.
Preferably, the keratin hydrolyzate has a mean molecular weight of less than 20'000 Da, preferably less than 10'000 Da, most preferably less than 5'000 Da. It has to be assumed that such keratin hydrolyzates lead to very dense, well-structured protective layers.
The molecular weight of the keratin hydrolyzate can be determined, for example, by SDS-PAGE method on tricine polyacrylamide gels in minicell Novex Model-3540 (USA) with constant voltage 125 V and current intensity 80 mA on start and 40 mA on end (Krejci et al, preparation and characterization of keratin hydrolyzates, Mathematical Methods and Techniques in Engineering and Environmental Science, ISBN: 978-1-61804-046-6).
Preferably, the protective layer has a thickness of 30 to 200 nm, most preferably of 30 to 90 nm. Interestingly, such thin layers can protect the hydrophilicity of the dental implant, but the roughness of the dental implant is maintained on the surface of the protective layer and results therefore in a better adherence. Therefore, a dental implant covered with a protective layer according to the present invention shows essentially the same surface roughness as a dental implant without said protective layer.
In fact, the density of the protective layer according to the present invention is remarkable. For both, a protective layer consisting of keratin hydrolyzate only and for one comprising keratin hydrolyzate together with a salt, the density of said layers is about 5% higher than a corresponding protective layer comprising glucose and a salt.
In one embodiment of the present invention, the protective layer additionally comprises a salt, preferably a bivalent salt, and most preferably MgCl2.
In a further embodiment of the present invention, the protective layer essentially free from any salts. Such a layer showed an outstanding homogeneity and was essentially free of cracks. In addition, the microroughness of the protective layer of the present invention is comparable with the microroughness of a non-coated dental implant. In fact, it is even difficult to distinguish between samples coated with the protective layer and the uncoated samples by scanning electron microscopy (SEM).
Furthermore, it was shown that the protective layer according to the present invention avoids the hydrothermal aging and keeps the hydrophilicity of the implant surface over time, preferably for at least one year, in particular of Y-TZP ceramic material.
Preferably, the dental implant comprising the protective layer according to the present invention is sterilized by ethylene oxide gas. This technique can be applied on the dental implant that is packed already. The ethylene oxide gas permeates the packaging and reaches the dental article to be sterilized. It could be shown that a sterilization by ethylene oxide gas does not negatively influence the stability of the protective layer. The hydrophilicity of the surface remains stable, which is particularly relevant when the dental implant is stored over relative long periods. In particular, when performing climatic stress over 8 weeks, the protective layer according to the present invention showed significant better hydrophilicity in comparison with a protective layer comprising, for example, agarose. Even after ethylene oxide (EO) sterilization, the implant surface coated by a protective layer according to the present invention was highly attractive for blood. Unprotected implants did not show the same affinity for blood which is in agreement with their hydrophobic nature.
Preferably, the protective layer is free of hydroxyapatite since protective layers that are free from hydroxyapatite have a higher density than the one comprising a combination of keratin hydrolyzate and hydroxyapatite.
Preferably, the dental implant is made of ceramic, preferably of an yttria stabilized zirconia ceramic. One of the main advantages of a zirconia implant is its natural white color that makes it undistinguishable from natural teeth. In addition, zirconia implants trigger a higher fibrinogen adsorption and a lower risk of bone loss. The protective layer shows similar features on a ceramic dental implant compared to an uncoated dental implant with the exception of a small decrease in the sharpness of the image, that is, there are smoother edges in the topography.
Alternatively, the dental implant is made of metal, preferably of titanium or a titanium alloy. A preferred titanium alloy is a binary titanium-zirconium alloy, and most preferably a binary titanium-zirconium alloy containing from 13 to 17 wt-%, more preferably from 13 to 15 wt-% of zirconium. This material has been shown to exhibit outstanding properties regarding mechanical stability and biofunctionality; a particularly preferred binary titanium-zirconium containing zirconium in the above-mentioned ranges is available under the trade-name Roxolid® (Institut Straumann AG, Switzerland). It was shown that the protective layer according to the present invention can also maintain the hydrophilicity of such an implant until the contact of the implant with the bone is established.
The dental implant according to the present invention can be constructed in one or more parts, in which case they consist of at least an anchoring part, often referred to in isolation as the implant, and an abutment, sometimes referred to as a spacer or post element. The anchoring part is usually either embedded completely in the bone, that is to say to the height of the alveolar crest (so called bone level implants), or protrudes by a few millimetres from the alveolar crest into the soft tissue (so called soft tissue level implants). The abutment is mounted either directly or indirectly to the anchoring part after the latter has become incorporated (osseointegrated) into the bone, or directly after the anchoring part has been inserted. It can also be attached to the anchoring part prior to insertion.
Preferably, such an anchoring part of a multi-part dental implant is generally in a cylindrical or conical shape, having an apical end with a body portion and a coronal end with a neck portion intended to receive an abutment in case of multi-part implants, and a transition portion being arranged in axial direction between the body portion and the neck portion. On said abutment a crown, a bridge or another suprastructure may be fixed, for example by screwing, cementing or gluing. The body portion is intended to be directed against the bone tissue in the implanted state and the neck portion is intended to direct against the soft tissue, whereas the transition portion may direct against the bone tissue or the soft tissue depending on the patient. Preferably, the total length of the anchoring part is 4 to 19 mm in the axial direction. The body portion with the apical end is preferably 50 to 90% of the total length of the implant in axial direction. The neck portion with the coronal end which is essentially be intended to be in contact with the soft tissue covers preferably 10 to 40% of the total length of the implant in axial direction. The transition portion covers preferably 0 to 40% of the total length of the implant in axial direction. The neck portion has preferably a length of 1 to 4 mm, most preferably 1.8 to 2.8 mm, the transition portion has preferably a length of 0 to 2 mm, most preferably 1 to 2 mm, and the body portion has preferably a length of 4 to 18 mm.
The body portion typically includes a threaded part which may be self-tapping or non-self-tapping.
Similar to the multi-part implant system, the anchoring portion of such a monotype implant is generally in a cylindrical or conical shape, having an apical end with a body portion and a coronal end with a neck portion passing into the abutment. The anchoring portion is typically provided with a threaded part, which may be self-tapping or non-self-tapping. A transition portion is arranged in axial direction between the body portion and the neck portion. On the abutment portion, a crown, a bridge or another suprastructure may be fixed, for example, by screwing, cementing or gluing. The body portion is intended to be directed against the bone tissue in the implanted state and the neck portion is intended to direct against the soft tissue, whereas the transition portion may direct against the bone tissue or the soft tissue depending on the patient.
The protective layer according to the present invention covers at least partially, preferably fully the hydrophilic surface of the dental implant and is preferably a continuous, crackless layer. In particular, if not only the body portion, but also the transition portion or the transition portion and the neck portion are covered with the protective layer, said protective layer is preferably one continuous layer.
In order to obtain an excellent osseointegration, the body portion is typically provided with a surface roughness. The term “surface roughness” within the context of the present invention stands for “arithmetical mean height” (Sa). Specifically, the surface roughness Sa (the arithmetic mean deviation of the surface in three dimensions) is determined in analogy to EN ISO 4287 relating to the respective parameters Ra in two dimensions. For the parameters in three dimensions, it is further referred to ISO 25178 (Sa is defined as Sq). Preferably, the surface roughness (Sa) is between 2 and 10 μm, preferably between 2 and 5 μm. Surprisingly, it is possible to maintain the surface roughness by the protective layer according to the present invention since the layer is extremely thin.
Preferably, the protective layer has a water content of less than 1% by weight. The absence of water to obtain a stable protective layer.
According to a further embodiment, the body portion is provided with a machined, but hydrophilic surface. Thus, said dental implants are acid etched, but no roughening step such as sandblasting, was applied.
Preferably, the dental implant coated with the protective layer is prepared by a comprising at least the following steps:
In order to providing the surface of the dental implant with a contact angle of less than 20°, the dental implant is preferably etchend by an inorganic acid or a blend of inorganic acids. More preferably, the inorganic acid(s) is/are selected from the group consisting of hydrofluoric acid, hydrochloric acid, sulphuric acid or mixtures thereof. Before carrying out the implant can at least partially be roughend mechanically and/or by using plasma technology and/or by laser structuring or by other methods known to the skilled person.
A preferred surface topography of an implant according to the present invention can be obtained for example by applying the process described in EP 1 982 670 or EP 1 982 671 before coating the roughened and etched surface. The disclosure of EP 1 982 670 and EP 1 982 671 is incorporated herein by reference.
Preferably, the surface of the dental implant is at least partly covered by dipping the implant into an aqueous solution or suspension comprising the keratin hydrolyzate. However, other methods to apply the aqueous solution to the surface to be protected are possible as well. Said dipping procedure ensures a constant thickness of the protective coating.
Preferably, the drying in step c), that is, the removal of water, is carried out by microwave treatment or by clear air drying (no specific drying, but storage at room temperature in a clean environment) drying in a convection oven, a vacuum oven or in an airstream, said airstream preferably having a temperature of 20° C. to 80° C., most preferably of 30° C. to 40° C. Preferably, the dental implants according to the present invention are dried in a convection oven at 20 to 80° C., preferably at 70° C., for 10 to 120 minutes, preferably for 15 to 90 minutes, most preferably for 20 to 60 minutes, ensuring that the protective layer is dried out completely. The above drying method does not induce splatters of the coating substance on the sample and allows to avoid hydrothermal aging.
Preferably, the hydrolyzed keratin is solubilized in an aqueous solution or aqueous suspension in a concentration of 0.1 to 10% by weight to volume (w/v), more preferably 1 to 5% by weight to volume (w/v).
The present invention is further illustrated by the following figures and examples:
The mounting part 220 has a frusto-conical or a conical shape and may be provided with at least one a flattening 230 at one side thereof.
At the side opposite the at least one flattening 260 there may be a groove 265 within the outer surface that extends from the coronal front surface of the mounting part 225 toward the apical side and ends in a conical section which forms the transition to the conical section of the anchoring part 205. The flattening 260 in combination with the groove 265 located on the opposite side functions to provide a positive a screwing tool which has a plug-in seat matched thereto. Alternatively, the mounting part may be provided with other means for receiving a screwing tool. The body portion 240 is intended to be directed against the bone tissue in the implanted state and the neck portion 220 is intended to direct against the soft tissue, whereas the transition portion 250 may direct against the bone tissue or the soft tissue depending on the patient. The surface of the body portion 240 has a contact angle of less than 20°, which is at least partly, preferably entirely covered with the protective layer 270. Optionally, not only the body portion 240 but also the transition portion 250 of the anchoring part is hydrophilic. Preferably, the transition portion 250 has a contact angle of less than 45°, preferably of less than 20°, which is covered by the protective layer 270. Preferably, at least 25%, most preferably at least 50% of the apical circumferential surface area of the transition portion is covered with said protective layer 250. This allows more flexibility when implanting the implant and ensures that the whole surface, which is intended to be in contact with the bone tissue, is hydrophilic.
The term ZLA within the context of the present invention stands for yttria stabilized zirconia, i.e. 3Y-TZP according to DIN ISO 12677, having a sand blasted (corundum 0.1-0.4 mm, 6 bar) and acid etched surface (for example HF). The used keratin hydrolyzate was Nutrilan® Keratin LM, BASF (CAS-Nr. 69430-36-0) with a molecular weight of 2000 g/mol. Ker:MgCl2 stands for a Nutrilan® Keratin LM:MgCl2 ratio of 9:1, whereby the keratin solution had a concentration of 1 mg/ml and the MgCl2 solution had a concentration of 0.05M.
Within the context of the present invention, “low concentration” stands for about 1 mg/ml (i.e. for example 1 mg Nutrilan® Keratin LM in 1 ml water which was obtained by adding 870 μl water per 100 ml of Nutrilan Keratin having a concentration of 115 mg/ml). The expression high concentration stands for about 30 mg/ml.
Samples used for coating experiments were cleaned and stored as follows:
Open the gas bottle and connect the O2 plasma cleaner and the valve to plasma. Press the pump button and wait until the pressure is lower than 8*10−2 mbar to switch on the gas. Give more gas flow (until 4-5*10−1 mbar) and wait 5 minutes. Lower the gas flow to 1*10−1 mbar and press the Generation button. Make sure that the parameters are set to 4 minutes (or two cycles of 2 minutes) and 35 W. Turn flow to 0 and switch off the gas. Stop pumping and open the chamber (Ventilation button). Take the samples holders from inside. Place the samples to be cleaned. Start cleaning procedure. Measure the contact angle of 3 samples to make sure that the process was successful (contact angle must be of a value of 0 degrees).
The sample was dipped in the dipping solution for 3 minutes on the coating beakers under sonication, placed on a Teflon mesh, placed in a shaker at 150 rpm for three minutes to get a more homogeneous coating and dried. The drying was carried out with recirculating air at 55° C. for 15 minutes in order to dry the coating on the samples.
Storage: the samples were stored in 24 well plates under laminar flow if no EO sterilization was performed.
Same procedure as for disks, with the following exceptions: No implants holder was used. They were submerged one by one in the coating solution and taken out with ceramic tweezers. Further, the implants were not placed in a Teflon mesh for trying, but directly in the implants primary package.
The experimental procedure was carried out according to DIN 55660-2:2011-12. Usually, the static contact angles were determined using a sessile drop test with ultrapure water (EasyDrop DSA20E, Kruss GmbH). The water droplets with a size of 0.1 μL for hydrophilic and 0.3 μL for hydrophobic discs of 5 mm diameter were dosed using an automated unit. Values for contact angles were calculated by fitting a circular segment function to the contour of the droplet placed on the surface.
Contact angles were determined for two different cases: (a) without washing the protection layer away and (b) after rinsing the samples with UPw for 15 seconds followed by blow drying in a stream of air in order to remove the coating.
As shown in
Further, in
Among the investigated protective layers at low concentrations only the discs coated with low-concentration keratin hydrolyzate could fulfil the criteria of having a contact angle below 25° without additional washing step performed by the clinical, ensuring sufficient hydrophilicity of the surface at the time of implantation (
Following the Wilhelmy plate method by means of a tensiometer (Lauda TE 3, Lauda Dr. R. Wobser GmbH & Co. KG), when a solid is submerged into a liquid and afterwards extracted from the liquid, the surface tension can be calculated from the necessary force that is needed for that specific solid.
The dynamic contact angle performed on coated ZLA implants showed that a coating with keratin hydrolyzate resulted in superhydrophilic implant surfaces, significantly outperforming their counterparts.
Effective microroughness values calculated using the 3D roughness values to be determined according to relevant procedure defined by Straumann Research with a low-pass Gaussian filter at 30 μm cut-off. Values of non-coated samples are used for all calculations, since that is the available surface for a coating.
The samples have to be dried before weight measurement (15 minutes in the oven at 55° C.; afterwards 110° C. for 40 minutes).
The weight difference of the discs before and after coating resulted in the following thickness estimation:
Since the samples have a specific micro surface roughness after sandblasting and acid etching, depending on the coating, the specific roughness parameters will change. These parameters are defined by the ISO 25178:
Ssk (AU): Skewness represents the symmetry of the surface heights in relation to the mean plane. If the parameter is larger than 0, peaks are predominant in front of valleys, while if the parameter is smaller than 0, valleys are predominant.
Sa (μm) and Sz (μm) Arithmetic mean height and maximum height represent a measure of the surface texture, containing information about the peaks and valleys, as well as information about the spacing in among those surface characteristics.
Sdr (μm): Developed interfacial area ratio represents the percentage of additional surface area that has to be added in relation to a complete flat surface without a texture.
Before analyzing the samples, they should be shortly rinsed under Argon gas in order to remove dust from the environment.
Parameters used: (objective lens: 20×/0.45—Mplan FL N; horizontal step distance: 0.22 μm; illumination: 70% exposure time/algorithm/quality: 28.5 ms/quick/max; gain: 1.5 dB).
In
Microroughness measurements of the coated implants, specifically Sa values, were compared between the apex (tip of the implant) and the 6th thread starting from the apex of the implant (
In order to check if the protective layers would be damaged under critical conditions during their storage, the cycle in Table 1 was applied after EO sterilization and the contact angle of the samples was measured and compared to the contact angle of the samples without going under this stress cycle.
As shown in
Aging parameters are established according to ASTM F1980-16
Accelerated Aging of Sterile Barrier Systems for Medical Devices.
At 2θ=30°, the higher intensity for the tetragonal phase was measured, with a depth of approximately 9.4 μm. The maximum depth penetration in the sample was about 20 μm, depending on the specific sample and if the mentioned was coated or not. The measure was performed from 10 to 40 incident ray degree
In order to test if the coatings provide antimicrobial activity, each sample was incubated in Lysogeny broth with E. coli bacteria. E. coli was pre-cultured one day before the infection start. At the time of the infection, a dissolution of the bacteria in fresh media to reach a concentration of 1*106 bacteria/mL were incubated (to test the amount of bacteria, Optical Density (OD) at 600 nm was measured and adjusted to 0.001). A control group was added, where no sample was added. From this control, the number of bacteria after 1 day is measured and compared to the samples of interest.
The bioburdent test consists of 7 days incubation of the samples in a LB media. Afterwards, OD-values of medium are taken to quantify the bacterial growth. If bacteria is observed, plating for CFU determination is needed.
Each protective layer was analyzed with three different replicates.
All samples were EO sterilized before analysis. The PLs were not removed, but cultivated in the media, so that sterilization was tested on the surface of the discs and in the dissolved components of the protective layers.
A caso broth was incubated at 30° C. in aerobic conditions during 14 days. Different media was used for each sample. Afterwards, visual analysis was performed to see if there were bacterial growth in the caso, according to ISO 11737-2. The analyzed microorganisms were the following:
In this case, one replicate was used for each PL.
As a proof of concept, the implants were dipped into blood. The blood was obtained fresh, and the implants were dipped into the blood for 10 seconds at the speed of 0.166 mm/second. After keeping for 6 minutes the implants in the blood (at 1.66 mm depth), the uncoated implant did not wet with blood media. Keratin layers quickly adsorbed blood, which resulted in an increasing quick red coloration of the threads from the apex upwards (after one minute it was already at the maximum blood adsorption).
Figure . . . Blood wettability of implants after 6 minutes made at the DCA instrument (at 1.66 mm depth from the first contact to the apex). (a) NoCoating (b) KerL
| Number | Date | Country | Kind |
|---|---|---|---|
| 20205274.2 | Nov 2020 | EP | regional |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/EP2021/080246 | 11/1/2021 | WO |
