CN112807488A - Ion adsorption type manganese dioxide coating with function of promoting bone differentiation and preparation method and application thereof - Google Patents
Ion adsorption type manganese dioxide coating with function of promoting bone differentiation and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses an ion adsorption type manganese dioxide coating with a function of promoting bone differentiation and a preparation method and application thereof. The ion adsorption type manganese dioxide coating with the function of promoting bone differentiation is an ion adsorption type manganese dioxide coating which grows on the surface of the base material in situ; the manganese dioxide coating is obtained by taking potassium permanganate as a manganese source, hydrochloric acid as a reducing agent and potassium chloride as a stabilizing agent and adopting a hydrothermal reaction method; wherein the crystal form of the manganese dioxide is birnessite type.
Description
Technical Field
The invention relates to an ion adsorption type manganese dioxide coating with a function of promoting bone differentiation, a preparation method and application thereof, belonging to the technical field of biomedicine.
Background
With the acceleration of the aging process of the population and the increase of bone injury accidents caused by traffic accidents, diseases, natural disasters and the like, the demand of artificial bone implant materials is increasing day by day. After the implant is implanted into a human body, the adsorption of proteins such as fibronectin on the surface of the implant can promote the actions of adhesion, proliferation, differentiation and the like of osteoblasts and improve the osseointegration capability. Researchers generally regulate factors such as the shape or chemical composition of the surface of an implant, so as to improve the fibronectin adsorption quantity and bone differentiation capacity of the implant. In a physiological environment, however, ions adsorb to the surface of the material before proteins. Therefore, ions adsorbed on the surface of the bone implant material can influence subsequent protein adsorption and cell behavior, and the research on a surface capable of regulating and controlling the concentration of adsorbed ions is of great significance.
Manganese dioxide (MnO)2) Is a transition metal oxide material with low price and easy preparation. Wherein the birnessite type MnO2The surface is rich in Mn vacancy, so that the surface has structural negative charge and can effectively adsorb metal ions. And the specific surface area of the lamellar structure is large, so that the adsorption and fixation of metal ions can be further promoted. Divalent calcium and magnesium ions adsorb more readily to MnO than monovalent sodium and potassium ions2Of (2) is provided. Further, Mn2+Is an important trace element in human body, directly participates in the synthesis of glycoprotein required by cartilage and bone matrix, and promotes the repair and regeneration of bones. Therefore, MnO having an ion adsorption function is prepared on the surface of the implant2The coating is expected to be used for regulating and controlling the adsorption behaviors of calcium and magnesium ions on the surface of the coating in a physiological environment, so that the fibronectin adsorption on the surface of an implant is promoted, and the possibility is provided for researching and developing a novel cheap biological coating for promoting the bone differentiation.
Disclosure of Invention
The present invention has been made to solve the above-mentioned problems occurring in the prior art, and an object of the present invention is to provide an ion-adsorbing MnO having a function of promoting bone differentiation2A coating and a preparation method and application thereof.
In a first aspect, the invention provides an ion-adsorbing manganese dioxide coating with a function of promoting bone differentiation, wherein the manganese dioxide coating is an ion-adsorbing manganese dioxide coating which grows in situ on the surface of a substrate; the manganese dioxide coating is obtained by taking potassium permanganate as a manganese source, hydrochloric acid as a reducing agent and potassium chloride as a stabilizing agent and adopting a hydrothermal reaction method; wherein the crystal form of the manganese dioxide is birnessite type.
Preferably, the substrate comprises a medical metal or medical alloy material, preferably at least one of pure titanium, titanium alloy, stainless steel or cobalt-chromium-molybdenum alloy.
Preferably, the concentration of the potassium permanganate is 0.001-0.01 mol/L.
Preferably, the concentration of the hydrochloric acid is 1-5 times of that of the potassium permanganate.
Preferably, the concentration of the potassium chloride is 1-5 times of that of the potassium permanganate.
Preferably, the hydrothermal reaction temperature is 80-150 ℃, and the heat preservation time is 1-24 h.
More preferably, the hydrothermal reaction temperature is 100-120 ℃, and the heat preservation time is 8-15 h.
The preparation method has the advantages of low cost, simple operation, good repeatability, suitability for large-scale production and the like.
The invention also provides an ion adsorption type manganese dioxide coating with the function of promoting bone differentiation, which is obtained by the preparation method.
Preferably, the manganese dioxide coating has an adsorption capacity of 0.5 to 12.0 μ g/cm for calcium ions2。
Preferably, the adsorption capacity of the manganese dioxide coating to magnesium ions is 0.1-5.0 mu g/cm2。
In a third aspect, the invention also relates to the use of an ion-adsorbing manganese dioxide coating having the function of promoting bone differentiation. In particular to application of a manganese dioxide coating for promoting fibronectin adsorption and preosteoblast osteogenic differentiation by regulating the adsorption behavior of calcium and magnesium ions on the surface of a material.
The manganese dioxide coating provided by the invention can effectively adsorb calcium and magnesium ions in a physiological environment, so that the adsorption quantity of fibronectin is improved, the alkaline phosphatase secretion and extracellular matrix mineralization of preosteoblasts are promoted, the expression of genes related to osteogenic differentiation is up-regulated, and the good osteogenic differentiation promoting capability is shown.
Drawings
FIG. 1 shows two MnO types, 5M100-Ti (hydrothermal reaction temperature 100 ℃ C., potassium permanganate concentration 0.005M) and 5M120-Ti (hydrothermal reaction temperature 120 ℃ C., potassium permanganate concentration 0.005M)2XRD spectrum of the coating.
FIG. 2 shows two MnO types of 5M100-Ti and 5M120-Ti2SEM pictures of the coating, wherein, the picture A is a SEM picture of 5M100-Ti coating multiplied by 3000, and the picture B is a SEM picture of 5M100-Ti multiplied by 30000; FIG. C is an SEM photograph of 5M120-Ti coating × 3000 times, and FIG. D is an SEM photograph of 5M120-Ti × 30000 times.
FIG. 3 shows two MnO types2Calcium ion (panel A) and magnesium ion (panel B) adsorption of the coatings (5M100-Ti and 5M 120-Ti).
FIG. 4 shows a Ti substrate and two MnO2Adsorption of the coatings (5M100-Ti and 5M120-Ti) on fibronectin in deionized water (containing fibronectin) and a calcium/magnesium ion mixed solution (containing fibronectin).
FIG. 5 shows preosteoblasts MC3T3-E1 in Ti substrate and two MnO2Cell proliferation (panel a), alkaline phosphatase activity (panel B) and extracellular matrix mineralization (panel C) of the surface of the coating (5M100-Ti and 5M 120-Ti).
FIG. 6 shows preosteoblasts MC3T3-E1 in Ti substrate and two MnO2Osteogenic differentiation-related genes were expressed on the surface of the coatings (5M100-Ti and 5M 120-Ti).
Detailed Description
The present invention is further illustrated by the following examples, which are to be understood as merely illustrative and not restrictive.
The ion-adsorbing manganese dioxide coating having osteogenic differentiation promoting function and the preparation method thereof according to the present invention will be described in detail below.
The coating with the function of promoting the bone differentiation provided by the invention,refers to ion adsorption type MnO growing on the surface of a substrate in situ2Coating of MnO2The crystal form of (A) is birnessite type. Compared with MnO of other crystal forms2Material of birnessite type MnO with lamellar structure2The specific surface area is large, the cation exchange capacity is high, the capability of adsorbing metal ions is particularly outstanding, and the coating is beneficial to the fixation of calcium and magnesium ions on the surface of the coating and the subsequent protein adsorption and osteoblast differentiation. The base material of the invention can adopt common medical metal or medical alloy materials such as pure titanium, titanium alloy, stainless steel or cobalt-chromium-molybdenum alloy and the like. Such a matrix material may provide better processability and mechanical properties.
The manganese dioxide coating can be obtained by taking potassium permanganate as a manganese source, hydrochloric acid as a reducing agent and potassium chloride as a stabilizing agent and adopting a hydrothermal reaction method. The invention adopts a hydrothermal reaction method which is simple to operate and can be produced in large scale, and MnO with excellent ion adsorption function is grown in situ on the surface of the base material2Coating to improve osteogenic differentiation capacity of bone implantation.
In some specific examples of the invention, the substrate is placed in a mixed solution of potassium permanganate, potassium chloride and hydrochloric acid, a hydrothermal reaction method is adopted, potassium permanganate is used as a manganese source, the concentration of the potassium permanganate in a reaction solution is 0.001-0.01 mol/L, hydrochloric acid is used as a reducing agent, potassium chloride is used as a stabilizing agent, and the hydrothermal reaction is carried out for 1-24 hours at 80-150 ℃. The concentration of the hydrochloric acid can be 1-5 times of that of the potassium permanganate. In some embodiments, the concentration of potassium chloride can be 1-5 times the concentration of potassium permanganate. Preferably, the hydrothermal reaction temperature is 100-120 ℃, and the heat preservation time is 8-15 h. In the hydrothermal process, if the reaction temperature is too high or the reaction time is too long, the lamellar structure of the birnessite type manganese dioxide collapses, so that the birnessite type manganese dioxide is converted into other forms of manganese dioxide, such as alpha-manganese dioxide. In the invention, potassium chloride is adopted as a stabilizer, potassium ions can balance the negative charge of the structure of the birnessite, the lamellar structure of the birnessite type is stabilized, and MnO is ensured2The material has larger specific surface area, which is beneficial to cation adsorption.
The manganese dioxide coating obtained by the method hasGood crystallinity, and a micro popcorn spherical structure formed by gathering the nano sheets. In some embodiments, the micro-popcorn spherical structures may have a size of 0.5 to 3 μm. The growth mechanism of the birnessite coating is as follows: in the early growth stage of the coating, potassium permanganate can generate oxidation-reduction reaction (4 MnO) with the titanium matrix4 -+3Ti+4H+→4MnO2+3TiO2+2H2O) to ensure that the manganese dioxide coating can grow in situ on the titanium substrate. In the later growth stage of the coating, potassium permanganate can generate oxidation-reduction reaction (2 MnO) with hydrochloric acid4 -+8H++6Cl-→2MnO2+3Cl2+4H2O), or self-decompose in the hydrothermal temperature range (2 MnO)4 -→MnO4 2-+MnO2+O2) Thereby forming MnO of a lamellar structure2And (4) coating. The morphology control mechanism in the Chinese patent 201711370891.X is that phosphate radical is adsorbed on the surface of birnessite (particularly the edge of a lamellar structure), so that MnO is realized2The directional growth of (2); if the phosphate radical concentration is lower, the flower ball-shaped lamellar structure is obtained.
The invention also discloses application of the ion adsorption type manganese dioxide coating with the function of promoting bone differentiation. The manganese dioxide coating prepared by the invention has good calcium and magnesium ion adsorption capacity, ions are adsorbed on the surface of the material before proteins in a physiological environment, fibronectin is easy to combine with divalent ions such as calcium, magnesium and the like, and calcium ion sites implanted on the surface of the material can promote the adsorption of fibronectin, thereby being beneficial to cell adhesion and osteogenic differentiation. In some embodiments, the manganese dioxide coating can have an adsorption capacity of 0.5 to 12.0 μ g/cm for calcium ions2. The manganese dioxide coating layer has an adsorption amount of magnesium ions of 0.1 to 5.0 [ mu ] g/cm2. These adsorbed calcium and magnesium ions serve as positive charge sites that promote the adsorption of negatively charged fibronectin on the surface of the material. Osteoblasts can specifically recognize fibronectin adsorbed on the surface of the material, so that osteogenic differentiation of cells on the surface of the material is promoted.
According to the invention, the adsorption performance of the birnessite type manganese dioxide is mainly utilized, and the component promoting function of the birnessite type manganese dioxide is realized after calcium and magnesium ions are adsorbed, so that the birnessite type manganese dioxide can be regarded as an expanded application of the adsorption function. The adsorption performance of manganese dioxide is positively correlated with its negative structural charge and the relative surface area of the coating. The potassium ions can balance the negative structural charge of the manganese dioxide, and the higher the content of the potassium ions in the coating is, the more negative charges need to be balanced in the material is shown, and the subsequent coating is favorable for exchange adsorption with calcium and magnesium ions. The larger the relative surface area of the coating, the more exposed the adsorption sites and the stronger the adsorption capacity of the coating.
The birnessite type MnO provided by the invention2The coating can effectively adsorb calcium and magnesium ions in a physiological environment, so that the adsorption quantity of fibronectin is improved, the secretion of alkaline phosphatase of preosteoblasts and the mineralization of extracellular matrix are promoted, the expression of genes related to osteogenic differentiation is up-regulated, and the coating has good capacity of promoting osteogenic differentiation. The preparation method has the advantages of low cost, simple operation, good repeatability, suitability for large-scale production and the like.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
Example 1
Preparation of MnO by hydrothermal reaction method2Coating layer
Placing the titanium sheet polished smooth by the abrasive paper in a mixed solution of potassium permanganate, potassium chloride and hydrochloric acid, and growing MnO on the surface of the titanium sheet in situ by adopting a hydrothermal reaction method2And (4) coating.
Wherein the volume of the ion mixed liquid is 60mL, the concentration of the potassium permanganate is 0.005M, the concentration of the potassium chloride is 0.015M, and the concentration of the hydrochloric acid is 0.02M. The hydrothermal reaction time is 12h, and the hydrothermal reaction temperature is 100 ℃ and 120 ℃. According to the concentration of the potassium permanganate and the hydrothermal reaction temperature, the obtained coating is marked as 5M100-Ti (the concentration of the potassium permanganate is 0.005M, and the hydrothermal reaction temperature is 100 ℃) and 5M120-Ti (the concentration of the potassium permanganate is 0.005M, and the hydrothermal reaction temperature is 120 ℃).
The coating was subjected to phase analysis. As shown in FIG. 1, the 5M100-Ti and 5M120-Ti coatings are both birnessite type MnO2(JCPDS 80-1098). Wherein MnO is contained in the 5M120-Ti coating2Degree of crystallinity ofMnO higher than 5M100-Ti in coating2The crystallinity of (a).
As shown in the SEM scanning picture of FIG. 2, the surfaces of the 5M100-Ti and 5M120-Ti coatings are all the popcorn spherical structures formed by the aggregation of the nanosheets, wherein the size of the popcorn spherical structures in the 5M120-Ti coating is larger.
Table 1 shows the relative surface area of the material measured by nitrogen adsorption, wherein MnO is2The relative surface area of the coating is significantly higher than that of the Ti substrate, and the relative surface area of the 5M120-Ti coating is larger than that of the 5M100-Ti coating.
TABLE 1 relative surface area of materials
| Relative surface area (cm)2/cm2) | |
| Ti | 26 |
| 5M100-Ti | 215 |
| 5M120-Ti | 263 |
Example 2
MnO2Calcium and magnesium ion adsorption amount detection of coating
The sample was immersed in a mixed solution of calcium chloride (1.8mM) and magnesium chloride (0.8mM) in a volume of 5mL for an adsorption time of 24 hours. And detecting the K/Mn ratio of the coating before and after adsorption by adopting photoelectron spectroscopy. And detecting the concentrations of calcium and magnesium ions in the adsorbed solution by adopting an inductively coupled plasma emission spectroscopy technology, and calculating the adsorption quantity of the calcium (magnesium) ions of the 5M100-Ti and 5M120-Ti coating samples.
As can be seen from Table 2, the content of exchangeable potassium ions in 5M120-Ti is higher than that in 5M100-Ti, indicating that the adsorption capacity of the 5M120-Ti coating is stronger. The change of the K/Mn ratio before and after adsorption shows that the amount of exchanged potassium ions of the 5M120-Ti coating is more.
TABLE 2K/Mn ratio Change before and after coating adsorption
| K/Mn (%, before adsorption) | K/Mn (%, after adsorption) | ΔK/Mn(％) | |
| 5M100-Ti | 10.8 | 2.7 | 8.1 |
| 5M120-Ti | 12.7 | 3.2 | 9.5 |
As shown in FIG. 3, the 5M100-Ti and 5M120-Ti coated samples were able to effectively adsorb calcium and magnesium ions, and the 5M120-Ti coated samples had better calcium and magnesium ion adsorption capacity. The hydrothermal reaction temperature is higher when the 5M120-Ti coating is prepared, and MnO is2The growth on the Ti substrate is faster, the required balance negative charge is more, and the exchangeable potassium ion content is moreHigh. Meanwhile, the laminated structure of the surface of the 5M120-Ti coating is more abundant, and the relative surface area is large. Therefore, the 5M120-Ti has stronger adsorption performance than the 5M 100-Ti.
Example 3
BCA (bicinchoninic acid) method for detecting fibronectin adsorption quantity of sample
The sample was immersed in deionized water or a mixed solution of calcium and magnesium ions (1.8mM calcium chloride and 0.8mM magnesium chloride) containing 40. mu.g/mL fibronectin in a volume of 1 mL. After 4h of adsorption, the supernatant was collected. 50 μ L of the supernatant was taken, and 200 μ L of BCA working solution was added and mixed well. The solution was incubated at 37 ℃ for 2h and the OD of the solution was measured by a microplate reader at 562 nm. And (4) drawing a standard curve according to the concentration of the protein standard substance and the corresponding OD value, and calculating the total protein concentration in the supernatant according to a standard equation. The amount of fibronectin adsorbed by the sample is the amount of change in the concentration of fibronectin in the solution before and after adsorption.
As shown in FIG. 4, the samples adsorbed 5M100-Ti > Ti on fibronectin in the presence and absence of calcium/magnesium ions. Wherein the adsorption amount of the Ti substrate to the fibronectin in the deionized water solution is larger than that of the Ti substrate to the fibronectin in the calcium/magnesium ion mixed solution. The adsorption capacity of the 5M100-Ti or 5M100-Ti coating sample on fibronectin in the calcium/magnesium ion mixed solution is larger than that of the sample on fibronectin in the deionized water solution. The calcium/magnesium ions are involved in the process of adsorbing fibronectin on the surface of the material, and the stronger the adsorption capacity of the sample on calcium and magnesium ions is, the more the sample can obviously improve the adsorption amount of fibronectin in the calcium/magnesium ion mixed solution.
Example 4
MnO2Detection of osteogenic differentiation-promoting Properties of the coating
Mouse preosteoblasts MC3T3-E1 are adopted to carry out experiments on cell proliferation, alkaline phosphatase activity, extracellular matrix mineralization and osteogenic differentiation related gene expression.
(1) Cell proliferation
The samples were sterilized using a steam sterilizer (121 ℃, 30min), and each set of sterile materials was carefully placed in a 48-well cell culture plate. Collecting growth stateGood MC3T3-E1 cells, digested and cell suspension concentration adjusted. 1mL of cell suspension (10000 cells/mL) was seeded onto the sample surface. At 37 deg.C, 5% CO2After culturing in the cell culture box for 1, 4 and 7 days, respectively, the culture solution was discarded. 1mL of fresh medium and 0.1mL of CCK-8 solution were added to each well. 37 ℃ and 5% CO2After further incubation for 3h in the cell incubator, the well solutions were carefully aspirated and added to the 96-well plates. The OD of each well was measured at 450nm using a microplate reader.
(2) Alkaline phosphatase Activity
The samples were sterilized using a steam sterilizer (121 ℃, 30min), and each set of sterile materials was carefully placed in a 48-well cell culture plate. The MC3T3-E1 cells with good growth state are collected, digested and the concentration of the cell suspension is adjusted. 1mL of cell suspension (50000 cells/mL) was seeded onto the sample surface. At 37 deg.C, 5% CO2After culturing for 7 and 14 days in the cell culture box, the culture medium was discarded. mu.L of 0.1% Triton X-100(PBS diluted) was added to each well. After the surface of the sample is blown by repeatedly sucking with a gun head, the liquid in the hole and the foam are sucked into an EP tube together for centrifugation (10000 revolutions, 5 min). Chromogenic substrate solutions and standard working solutions (0.5mM) were prepared, and blank control wells, standard wells, and sample wells were set using 96-well plates with reference to Table 1. The amounts of standards were 4, 8, 16, 24, 32 and 40 μ L, respectively. After mixing the liquid with the aid of a shaker (50rpm/min), incubation was carried out for 10min at 37 ℃. mu.L of stop solution was added to each well, and absorbance was measured at 405nm using a microplate reader. ALP quantification was obtained after normalization of total protein concentration.
TABLE 3 arrangement of blank control well, standard well and sample well
| Blank control | Standard article | Sample (I) | |
| Detection buffer solution | 50μL | 100-x | - |
| Chromogenic substrates | 50μL | - | 50 |
| Sample (I) | - | - | 50 |
| Working solution for standard substance | - | x | - |
(3) Extracellular matrix mineralization
The samples were sterilized using a steam sterilizer (121 ℃, 30min), and each set of sterile materials was carefully placed in a 48-well cell culture plate. The MC3T3-E1 cells with good growth state are collected, digested and the concentration of the cell suspension is adjusted. 1mL of cell suspension (50000 cells/mL) was seeded onto the sample surface. At 37 deg.C, 5% CO2After 14 and 21 days of culture in the cell incubator, respectively, the culture medium was discarded. 1mL of 4% paraformaldehyde solution was added to each well and incubated at 4 ℃ for 15 min. The supernatant was discarded, washed with PBS 3 times, and 500. mu.L of alizarin red dye solution was added to each well, followed by incubation at room temperature for 20 min. The supernatant was discarded, washed with PBS 3 times, and 500. mu.L of 10% cetylpyridinium chloride solution was added to each well at room temperatureIncubate for 15 min. OD was measured at 590nm using a microplate reader.
(4) Osteogenic differentiation-related Gene expression
The samples were sterilized using a steam sterilizer (121 ℃, 30min), and each set of sterile materials was carefully placed in a 48-well cell culture plate. MC3T3-E1 cells were separately digested, counted, and formulated into cell suspensions at a concentration of 250000/mL. 2mL of cell suspension was seeded onto the surface of each well material. After standing for 6h, the supernatant was discarded and replaced with osteoinductive fluid. At 37 ℃ with 5% CO2And (5) culturing in an incubator for 7 d. And detecting the expression of the genes related to osteogenic differentiation of the MC3T3-E1 cells by adopting a qPCR method.
As can be seen from FIG. 5, the proliferation behaviors of MC3T3-E1 preosteoblasts on the surfaces of 5M100-Ti and 5M120-Ti coatings are not obviously different from that on the surface of a Ti substrate, which indicates that the developed coating has good biocompatibility. The 5M100-Ti and 5M120-Ti coated samples were significantly better than the Ti substrate control group in their ability to promote differentiation of preosteoblasts (alkaline phosphatase activity) and mineralization of extracellular matrix of MC3T 3-E1. And the 5M120-Ti coating sample has better capability of promoting differentiation and mineralization of osteoblasts before MC3T3-E1 than the 5M100-Ti coating sample.
As can be seen from FIG. 6, the 5M100-Ti and 5M120-Ti coated samples have significantly better ability to promote the expression of the osteogenic differentiation associated genes (Col1, Opn, Bmp2 and Runx2) in cells than the Ti substrate control group. And the 5M120-Ti coated sample has better capability of promoting the expression of genes (Col1, Opn, Bmp2 and Runx2) related to the osteogenic differentiation of cells than the 5M100-Ti coated sample.
Claims (10)
1. An ion adsorption type manganese dioxide coating with the function of promoting bone differentiation, which is characterized in that the manganese dioxide coating is an ion adsorption type manganese dioxide coating growing on the surface of a base material in situ; the manganese dioxide coating is obtained by taking potassium permanganate as a manganese source, hydrochloric acid as a reducing agent and potassium chloride as a stabilizing agent and adopting a hydrothermal reaction method; wherein the crystal form of the manganese dioxide is birnessite type.
2. Manganese dioxide coating according to claim 1, wherein said substrate comprises a medical metal or medical alloy material, preferably at least one of pure titanium, titanium alloy, stainless steel or cobalt chromium molybdenum alloy.
3. The method for preparing manganese dioxide coating according to claim 1 or 2, wherein the concentration of potassium permanganate is 0.001-0.01 mol/L.
4. The method according to any one of claims 1 to 3, wherein the concentration of the hydrochloric acid is 1 to 5 times that of potassium permanganate.
5. The method according to any one of claims 1 to 4, wherein the concentration of the potassium chloride is 1 to 5 times that of the potassium permanganate.
6. The preparation method according to any one of claims 1 to 5, wherein the hydrothermal reaction temperature is 80 to 150 ℃ and the holding time is 1 to 24 hours.
7. The preparation method according to claim 6, wherein the hydrothermal reaction temperature is 100-120 ℃ and the holding time is 8-15 h.
8. Ion-adsorbing manganese dioxide coating with a function of promoting bone differentiation obtained by the preparation method according to any one of claims 1 to 7.
9. Use of an ion-adsorbing manganese dioxide coating according to claim 8 to promote osteogenic differentiation.
10. The use according to claim 9, wherein said manganese dioxide coating promotes fibrin adsorption and osteoblastic osteogenic differentiation by modulating the adsorption of calcium and magnesium ions on its surface.
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