NANO calcium phosphate surfaces

Abstract

The present disclosure relates to a calcium phosphate coating that includes a plurality of nanometer sized calcium phosphate crystals that form a single layer. The coating is on a surface of a conductive substrate and is about 1 to 1000 nanometers thick, and greater than 25 percent of the coated surface area comprises the single layer of the crystals.

Description

Docket No. 2891.098AWO NANO CALCIUM PHOSPHATE SURFACES FIELD

[0001] The present disclosure relates to nanometer thick calcium phosphate surface coatings. BACKGROUND

[0002] Calcium phosphate coatings have many uses including coating metal surfaces in medical implants. Calcium phosphate coatings on medical implant surfaces are commonly created using a plasma spray method resulting in a calcium phosphate coating that is many microns in thickness. The micron thick calcium phosphate layer is thick enough to act independently from the surface of the implant leading to delamination. Micron thick calcium phosphate coatings include a surface roughness or texture that is also microns in size and may not promote cellular interactions with the coating. The present disclosure is directed to overcoming these and other limitations. SUMMARY

[0003] In an aspect, disclosed is a calcium phosphate coating comprising a plurality of nanometer sized calcium phosphate crystals, wherein the crystals form a single layer, wherein the coating is on a surface of a conductive substrate and is about 1 to 1000 nanometers thick, and wherein greater than 25 percent of the coated surface area comprises the single layer of the crystals.

[0004] In a further aspect, the conductive substrate is metal. In a further example, greater than 50 percent of an exterior surface comprises the single layer of the crystals. In yet a further example, greater than 50 percent of the crystals are vertically orientated about 20 degrees or greater from the surface. In still a further example, the crystals are ionically bound to the substrate.

[0005] In an example, the crystals are created from a layer of amorphous calcium phosphate 1 to 1000 nanometers thick on the substrate. In a further example, about 75 percent of the crystals are vertically orientated about 20 degrees or greater from the surface. In a yet further example, about 95 percent of the crystals are vertically orientated about 20 degrees or greater from the surface.

[0006] In an example, a density of the crystals is determined by a thickness of the amorphous calcium phosphate coating. In a further example, a size of the crystals is determined by a rate of conversion of amorphous calcium phosphate to crystalline calcium phosphate and a totalDocket No. 2891.098AWO conversion time. In yet a further example, the crystals are about 1 – 500 nanometers in length. In still a further example, the crystals have an aspect ratio of about 1:1.5 to 1:10. In an example, the coating is about 1 to about 100 nanometers thick. In a further example, the coating is about 150 to about 1000 nanometers thick. In yet a further example, the coating is about 100 to about 500 nanometers thick. In still a further example, the coating is about 500 to about 1000 nanometers thick.

[0007] In an example, the calcium phosphate includes hydroxyapatite, calcium deficient hydroxyapatite, octacalcium phosphate, dicalcium phosphate dihydrate, tricalcium phosphate and brushite.

[0008] In an example, the coating is adhered to the substrate at less than 100 degrees centigrade. In a further example, the calcium phosphate coating is adhered to the substrate by electrodeposition in an aqueous solution. In yet a further example, the calcium phosphate coating is adhered to the substrate by nanosecond electric pulse electrodeposition in an aqueous solution. In still a further example, the substrate includes an engineered nanometer scale topology and wherein the calcium phosphate coating preserves the nanometer scale topology of the substrate. In an example, the substrate includes an engineered micron scale topology and wherein the calcium phosphate coating preserves the micron scale topology of the substrate.

[0009] In an example, one or more dopants are adhered to the substrate with the calcium phosphate coating. In a further example, one or more dopants are electrodeposited with the calcium phosphate coating to substrate. In a yet further example, one or more dopants are diffused into the calcium phosphate coating. In still a further example, nanoparticles are added interstitial to a calcium phosphate layer or layers. In an example, nanoparticles are co-deposited with the calcium phosphate coating onto the substrate. In a further example, the conductive substrate comprises nanoparticles prior to addition of the calcium phosphate coating. In yet a further example, nanoparticles are added to the coating after the calcium phosphate coating.

[0010] In an example, one or more dopants comprise copper, magnesium, manganese, strontium, zinc, potassium, silver, gold, sodium, fluoride, chlorine, bromine, ions of the foregoing, or any combination thereof. In a further example, the calcium phosphate coating dissolves at a predetermined time, rate or combinations of both. In yet a further example, the calcium phosphate coating dissolves at a predetermined time to release one or more dopant, nanoparticleDocket No. 2891.098AWO or combinations of both. In still a further example, the calcium phosphate coating dissolves at a predetermined rate to release one or more dopant, nanoparticle or combinations of both.

[0011] In an example, one or more dopants comprise DNA, RNA, vitamins, minerals, antimicrobials, antivirals, steroids, anti-inflammatories, pain relief medication and growth factors. In a further example, the nanoparticles comprise DNA, RNA, vitamins, minerals, antimicrobials, antivirals, steroids, anti-inflammatories, pain relief medication and growth factors. In yet a further example, dissolution of the calcium phosphate coating releases antimicrobials. In still a further example. at least one additional amorphous calcium phosphate layer is deposited on the coating. In an example, at least one additional amorphous calcium phosphate layer comprising dopants is deposited on the coating. In a further example, at least one additional amorphous calcium phosphate layer comprising biological agents is deposited on the coating. In a yet further example, at least one additional amorphous calcium phosphate layer dissolves after a predetermined period of time and the calcium phosphate coating dissolves at a second predetermined period of time after the amorphous calcium phosphate layer. In still a further example, one or more dopant deposited with the calcium phosphate determines a dissolution rate of one or more coating.

[0012] In an example, the coating promotes mesenchymal stem cell differentiation into osteoblasts. In a further example, the coating includes at least one dopant which further promotes mesenchymal stem cell differentiation into osteoblasts as compared to the calcium phosphate coating alone.

[0013] In an example, the coating contains one or more fluorescent dye, photoluminescent dye, radiochemical or combinations thereof. In a further example, the coating comprises a first nanometer thick layer of amorphous calcium phosphate coated on the conductive substrate, wherein the amorphous calcium phosphate coating is converted to crystalline calcium phosphate using a solution with a basic pH. In a yet further example, the coating includes a first nanometer thick layer of amorphous calcium phosphate coated on the conductive substrate, wherein the amorphous calcium phosphate coating is converted to crystalline calcium phosphate using a solution with a basic pH, wherein the crystalline calcium phosphate comprises nanometer sized calcium phosphate crystals and wherein the size of the plurality of nanometer calcium phosphateDocket No. 2891.098AWO crystals is determined a pH of the solution, a temperature of the solution and a time the first nanometer thick layer of amorphous calcium phosphate soaks in the solution.

[0014] In an aspect, disclosed is an amorphous calcium phosphate coating, wherein the coating is on a surface of a conductive substrate and is about 1 to 1000 nanometers thick, and wherein the coating is characterized by a nanometer surface roughness wherein the nanometer surface roughness can come from a roughness of the conductive substrate, a roughness of the amorphous calcium phosphate coating or a combination of both. In an example, the amorphous calcium phosphate is deposited on the conductive substrate using nanosecond electric pulses. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] The accompanying drawings, which are incorporated in and constitute a part of the specification, illustrate embodiments and examples of the disclosure and together with the detailed description herein, serve to explain the principles of the disclosure. The drawings are only for purposes of illustrating embodiments and are not to be construed as limiting the disclosure. It is emphasized that, in accordance with the standard practice in the industry, various features may not be drawn to scale. In fact, the dimensions of the various features may be arbitrarily increased or reduced for clarity of discussion. These and other features, aspects, and advantages of the present disclosure will become better understood when the following detailed description is read with reference to the accompanying drawings, wherein:

[0016] FIGS. 1A and 1B shows a non-limiting embodiment of cathodic deposition of calcium phosphate on a stainless steel washer, in accordance with an aspect of the present disclosure;

[0017] FIG. 2 shows a non-limiting embodiment of the measured output of a device generating nanosecond pulses of electric current for electrodeposition of calcium phosphate coatings, in accordance with an aspect of the present disclosure;

[0018] FIG. 3 shows a non-limiting embodiment of amorphous calcium phosphate on a stainless steel washer, in accordance with an aspect of the present disclosure;

[0019] FIG. 4 shows a non-limiting embodiment of an SEM image of crystalline calcium phosphate, in accordance with an aspect of the present disclosure;Docket No. 2891.098AWO

[0020] FIG. 5 shows a non-limiting embodiment of amorphous calcium phosphate on a stainless steel washer, in accordance with an aspect of the present disclosure;

[0021] FIG. 6 shows a non-limiting embodiment of an SEM image of crystalline calcium phosphate, in accordance with an aspect of the present disclosure;

[0022] FIG. 7 shows a non-limiting embodiment of a SEM image of a control stainless steel surface without any coating, in accordance with an aspect of the present disclosure;

[0023] FIG. 8 shows a non-limiting embodiment of an SEM image of amorphous calcium phosphate on a stainless steel surface, in accordance with an aspect of the present disclosure;

[0024] FIG. 9 shows a non-limiting embodiment of an SEM image of calcium phosphate crystals, large and small, converted using basic pH solution, in accordance with an aspect of the present disclosure;

[0025] FIG. 10 shows a non-limiting embodiment of an SEM image of nanoscale topology of TiO2 Nanotubes without any coating, in accordance with an aspect of the present disclosure;

[0026] FIG. 11 shows a non-limiting embodiment of an SEM image of amorphous Zn-CaP on TiO2 nanotubes, in accordance with an aspect of the present disclosure;

[0027] FIG. 12 shows a non-limiting embodiment of an SEM image of crystalline converted Zn- CaP on TiO2 nanotubes, in accordance with an aspect of the present disclosure;

[0028] FIG. 13 shows a non-limiting embodiment of a graph quantifying Alizarin red on various titanium surfaces after 21 day differentiation of MSCs, in accordance with an aspect of the present disclosure;

[0029] FIG. 14 shows a non-limiting embodiment of a graph quantifying Alizarin red on various stainless steel surfaces after 21 day differentiation of MSCs, in accordance with an aspect of the present disclosure;

[0030] FIG. 15 shows a non-limiting embodiment of calcium phosphate crystal vertical orientation as compared to the surface of the substrate, in accordance with an aspect of the present disclosure;Docket No. 2891.098AWO

[0031] FIG. 16 shows non-limiting embodiments of ions used as dopants including embodiment concentration ranges and embodiments of biological effects, in accordance with an aspect of the present disclosure;

[0032] FIG. 17 shows a non-limiting embodiment of low density calcium phosphate crystals, in accordance with an aspect of the present disclosure;

[0033] FIG. 18 shows a non-limiting embodiment of high density calcium phosphate crystals, in accordance with an aspect of the present disclosure;

[0034] FIG. 19 shows a non-limiting embodiment of a nanometer thick crystalline calcium phosphate coating on a stainless steel surface, in accordance with an aspect of the present disclosure. DETAILED DESCRIPTION

[0035] This disclosure relates to nanometer thick calcium phosphate (CaP) coatings on conductive substrates where the coating includes a single layer of nanocrystals that have a vertical orientation relative to the surface of the substrate. The nanocrystals may exhibit 22 degrees or more of a vertical orientation relative to the substrate. The nanocrystals may be highly ordered and impart new properties to the underlying substrate. The nanocrystal density may be increased or decreased. The nanocrystals may bind to the substrate through ionic bonding or mostly through ionic bonding resulting in a strong attachment resistant to delamination and physical stress. The nanometer thick coatings may be applied by nanosecond pulses of direct electric current using electrodeposition. In some embodiments, a conductive substrate is placed in an electrolyte wherein the calcium phosphate is provided in an ionic form, such as one or more salts, in the electrolyte and is electrochemically deposited on the conductive substrate during the nanosecond electric pulses. The calcium phosphate coatings may be electrodeposited at temperatures that retain the function of co-deposited biopolymers and other compounds that may be heat sensitive., The calcium phosphate coating is initially electrochemically deposited as a nanometer thick amorphous calcium phosphate (ACP) coating and then can be converted to crystalline calcium phosphate using a basic or alkaline solution. The resulting crystalline phosphate coating maintains the nanometer thickness. The conversion process does not add anyDocket No. 2891.098AWO additional calcium phosphate but converts nanometer thick amorphous calcium phosphate to crystalline calcium phosphate. In some embodiments, the crystalline calcium phosphate is hydroxy apatite (HAP). In another embodiment the crystalline calcium phosphate is octacalcium phosphate (OCP). The ratio of calcium to phosphorous may be modified to produce a variety of calcium phosphate coatings with different physical and biological characteristics. Dopants, additional materials that may be incorporated into the calcium phosphate coating, may be included to impart new properties as described herein.

[0036] When describing nanometer thick amorphous, crystalline and semicrystalline calcium phosphate the term calcium phosphate may be used as a way to describe each form of calcium phosphate and characteristics that may be shared by each form such as thickness and dopants. In some embodiments, the specific morphology of calcium phosphate such as amorphous, crystalline and semicrystalline will be specified. This disclosure teaches a highly uniform calcium phosphate coating that is nanometers in thickness with new surface properties. All calcium phosphate coatings disclosed herein are nanometers thick and nanometer scale. The nanometer thick calcium phosphate coatings may contain nanometer size features. Nanometer size features and nanometer thick coatings may be referred to as nanometer scale indicating that one or more feature of the calcium phosphate coating has dimensions on the 1 to 1000 nanometer scale. Nanometer size, nanometer, nanometer thick, nanometer thickness and nanometer scale may include dimensions of calcium phosphate coatings such as thickness, crystal dimensions such as crystal height, crystal width and crystal depth, crystal topology, substrate topology, roughness, substrate features and nanoparticles where at least one measurement is about 1 to 1000 nanometers. Roughness may be calculated as average roughness, Ra, using the average, or arithmetic average of profile height deviations from the mean line. Roughness may be calculated using the quadratic mean, or root mean square average of profile height deviations from the mean line as Rq. Roughness may be calculated as the average peak height to the surface, such that a roughness of 100 nanometers means the average height of a feature, such as crystal, dopant, nanoparticle, nanotube, surface feature, surface texture, is on average 100 nanometers in height from the surface. A nanometer surface roughness may include a surface of features that are nanometers in size. A micron surface roughness may include a surface of features that are microns in size.Docket No. 2891.098AWO

[0037] The disclosed crystalline calcium phosphate coating is nanometers in thickness and the calcium phosphate crystals are nanometers in at least one dimension. The nanometer size crystals may be formed from a highly uniform single layer of amorphous calcium phosphate that is nanometers in thickness and converted with a basic solution to a highly uniform single layer of nanometer size crystalline calcium phosphate. The reduction in size of calcium phosphate crystals from micrometer size to nanometer size is significant and results in new properties in the calcium phosphate coating not found in micrometer thick coatings.

[0038] The disclosed nanometer scale crystalline calcium phosphate coating includes nanometer sized crystals that have a vertical orientation as compared to the surface of the substrate. Nanocrystal vertical orientation may be an angle measured from a central axis of a crystal to the surface of the substrate. In some embodiments a vertical orientation of 90 degrees from the surface of the substrate represents the greatest measured angle from the surface of the substrate. In some embodiments having a maximum 90-degree vertical orientation allows all crystal vertical orientation to be described in angles of 90 degrees or less from the surface of the substrate regardless of the direction of the crystal. Thus, where crystals face north, south, east or west, where north is the top of the image, the vertical orientation will be described relative to a perpendicular angle to the surface of the substrate where 90 degrees represents the greatest vertical orientation relative to the surface of the substrate.

[0039] A vertical orientation and vertically oriented may be used to describe an angle from a central axis of a crystal to the surface of a substrate. In some embodiments a vertical orientation expressed as an angle from a central axis of a nanocrystal to the surface of the substrate represents an average vertical orientation of each calcium phosphate nanocrystal of the plurality of nanocrystals. In some embodiments a plurality of calcium phosphate nanocrystals may be described by a minimum vertical orientation, expressed by an angle from the surface of the substrate to a central axis of a plurality of crystals wherein each nanocrystal of the plurality of nanocrystals exceeds the minimum vertical angle. In some embodiments, nanometer size crystals are vertically oriented at least 20 or more degrees from the surface. In some embodiments the calcium phosphate crystals are vertically oriented at least 22.5 degrees vertical from the surface.

[0040] The vertical nature of the crystals on the disclosed calcium phosphate coatings imparts new physical properties into the coating. The disclosed vertical crystals result in more freeDocket No. 2891.098AWO bonding atoms available to react with the environment. In some embodiments, crystals project nanometers from the substrate surface and create a novel vertical crystal topology of available binding atoms.

[0041] In some embodiments, a percentage of the plurality of nanometer sized calcium phosphate crystals are vertically oriented. In some embodiments, a percentage of the plurality of nanometer sized calcium phosphate crystals have a vertical orientation. In some embodiments the percentage of the plurality nanometer calcium phosphate crystals with a vertical orientation may be about 30 percent, about 40 percent, about 50 percent, about 55 percent, about 60 percent, about 65 percent, about 70 percent, about 75 percent, about 80 percent, about 85 percent, about 90 percent, about 95 percent or about 100 percent.

[0042] In some embodiments, a percentage range of the plurality of nanometer sized calcium phosphate crystals are vertically oriented. In some embodiments, the percentage range of the plurality nanometer calcium phosphate crystals vertically oriented may be about 30 percent or greater , about 40 percent or greater, about 50 percent or greater, about 55 percent or greater, about 60 percent or greater, about 65 percent or greater, about 70 percent or greater, about 75 percent or greater, about 80 percent or greater, about 85 percent or greater, about 90 percent or greater, about 95 percent or greater or about 100 percent. In describing range boundaries “or greater” may include about 50 percent, about 60 percent, about 70 percent, about 80 percent, about 90 percent or about 100 percent.

[0043] In some embodiments a percentage of the surface of the conductive substrate is coated with a single layer of calcium phosphate nanocrystals. In some embodiments a surface includes an exterior surface of conductive substrate. In some embodiments a surface includes an internal surface of a conductive substrate. In some embodiments a surface includes an internal surface, an exterior surface, or a combination of both surfaces. In some embodiments a porous surface results in an external surface of a conductive surface having a greater percentage of the surface coated with a single layer of calcium phosphate nanocrystals than an interior surface. In some embodiments a percentage of a surface coated with a single layer of calcium phosphate nanocrystals may be about 1 percent, about 5 percent, about 10 percent, about 15 percent, about 20 percent, about 25 percent, about 30 percent, about 40 percent, about 50 percent, about 55Docket No. 2891.098AWO percent, about 60 percent, about 65 percent, about 70 percent, about 75 percent, about 80 percent, about 85 percent, about 90 percent, about 95 percent or about 100 percent.

[0044] In some embodiments a minimum percentage of a surface of a conductive substrate is coated in calcium phosphate nanocrystals. In some embodiments a surface of a conductive substrate may be covered by about 5 percent or greater, about 10 percent or greater, about 15 percent or greater, about 20 percent or greater, about 25 percent or greater, about 30 percent or greater , about 40 percent or greater, about 50 percent or greater, about 55 percent or greater, about 60 percent or greater, about 65 percent or greater, about 70 percent or greater, about 75 percent or greater, about 80 percent or greater, about 85 percent or greater, about 90 percent or greater, about 95 percent or greater or about 100 percent. In describing range boundaries “or greater” may include about 50 percent, about 60 percent, about 70 percent, about 80 percent, about 90 percent or about 100 percent.

[0045] In some embodiments, a plurality of calcium phosphate nanocrystals may be vertically orientated where the vertical orientation includes a minimum vertical angle. A minimum vertical angle may include a minimum angle exceeded by each crystal. A minimum vertical angle may include a minimum angle wherein each crystal meets or exceeds the minimum vertical angle. In some embodiments, a plurality of calcium phosphate crystals are vertically oriented with a minimum vertical angle of about 10 degrees, about 15 degrees, about 20 degrees, about 22.5 degrees, about 25 degrees, about 30 degrees, about 35 degrees, about 40 degrees, about 45 degrees, about 50 degrees, about 55 degrees, about 60 degrees, about 65 degrees, about 70 degrees, about 75 degrees, about 80 degrees, about 85 degrees or about 90 degrees from the surface of the substrate.

[0046] In some embodiments a plurality of calcium phosphate nanocrystals may be vertically orientated where a vertical orientation is measured from the surface of a substrate to a central axis of each of a plurality of nanocrystals and a vertical orientation represents an average vertical orientation of the plurality of nanocrystals. In some embodiments a plurality of calcium phosphate nanocrystals are vertically oriented about 10 degrees, about 15 degrees, about 20 degrees, about 22.5 degrees, about 25 degrees, about 30 degrees, about 35 degrees, about 40 degrees, about 45 degrees, about 50 degrees, about 55 degrees, about 60 degrees, about 65Docket No. 2891.098AWO degrees, about 70 degrees, about 75 degrees, about 80 degrees, about 85 degrees or about 90 degrees from the surface of the substrate.

[0047] In some embodiments crystal vertical orientation is determined based on images. In some embodiments crystal vertical orientation is determined using computer software analysis of the images.

[0048] The nanometer calcium phosphate crystals may have an aspect ratio that describes the length of the crystal as compared to the width. An aspect ratio of 1 unit of length to 1 unit of width is 1:1 and would be spherical in shape. An aspect ratio may be the shortest dimension to the longest dimension. In some embodiments, the width may be the dimension that is the shortest dimension and the length the longest dimension. In some embodiments, the length is measured as the distance the crystal extends from the surface. In some embodiments, the aspect ratio of the nanometer calcium phosphate crystal can be controlled during the conversion process. In some embodiments, a crystal with a higher ratio than 1:1 such as 1:2 or 1:3 may be created using a weaker base that is cooled. In some embodiments, a crystal aspect ratio that is lower than 1:2 or such as 1:1.2 or 1:1.3 may be created using a stronger base that is heated. The disclosed nanometer scale calcium phosphate crystals may have aspect ratios of about of about 1:1.2, of about 1:1.3, of about 1:1.4, of about 1:1.5, of about 1:1.6, of about 1:1.7, of about 1:1.8, of about 1:1.9, of about 1:2, of about 1:2.1, of about 1:2.2, of about 1:2.3, of about 1:2.4, of about 1:2.5, of about 1:2.6, of about 1:2.7, of about 1:2.8, of about 1:2.9, of about 1:3, of about 1:3.1, of about 1:3.2, of about 1:3.3, of about 1:3.4, of about 1:3.5, of about 1:3.6, of about 1:3.7, of about 1:3.8, of about 1:3.9, of about 1:4, of about 1:4.1, of about 1:4.2, of about 1:4.3, of about 1:4.4, of about 1:4.5, of about 1:4.6, of about 1:4.7, of about 1:4.8, of about 1:4.9, of about 1:5, of about 1:5.1, of about 1:5.2, of about 1:5.3, of about 1:5.4, of about 1:5.5, of about 1:5.6, of about 1:5.7, of about 1:5.8, of about 1:5.9, of about 1:6, of about 1:6.1, of about 1:6.2, of about 1:6.3, of about 1:6.4, of about 1:6.5, of about 1:6.6, of about 1:6.7, of about 1:6.8, of about 1:6.9, of about 1:7, of about 1:7.1, of about 1:7.2, of about 1:7.3, of about 1:7.4, of about 1:7.5, of about 1:7.6, of about 1:7.7, of about 1:7.8, of about 1:7.9, of about 1:8, of about 1:8.1, of about 1:8.2, of about 1:8.3, of about 1:8.4, of about 1:8.5, of about 1:8.6, of about 1:8.7, of about 1:8.8, of about 1:8.9, of about 1:9, of about 1:9.1, of about 1:9.2, of about 1:9.3, of about 1:9.4, of about 1:9.5, of about 1:9.6, of about 1:9.7, of about 1:9.8, of about 1:9.9, or of about 1:10.Docket No. 2891.098AWO

[0049] The nanometer size calcium phosphate crystals may be described in terms of the crystal space group or family and may be monoclinic, triclinic, rhombohedral, hexagonal or combinations thereof. In some embodiments, the crystal structure is uniform in the coating. In some embodiments, the nanometer crystalline calcium phosphate coating includes a single layer of monoclinic nanometer crystals. In some embodiments, the nanometer crystalline calcium phosphate coating includes a single layer of triclinic nanometer crystals. In some embodiments, the nanometer crystalline calcium phosphate coating includes a single layer of rhombohedral nanometer crystals. In some embodiments, the nanometer crystalline calcium phosphate coating includes a single layer of hexagonal nanometer crystals. In some embodiments, the nanometer crystalline calcium phosphate coating includes a single layer of one or more monoclinic, triclinic, rhombohedral, and hexagonal crystals.

[0050] Nanometer thick coatings of amorphous calcium phosphate may be electrodeposited on electrically conductive substrates in varying ratios of calcium to phosphate. The calcium phosphate coating may have many chemical configurations. The ratio of calcium to phosphorous may be expressed as a ratio in a number of ways such as calcium : phosphorous and calcium / phosphorous. In some embodiments, the ratio of calcium to phosphorus can vary from 1:1 to 2:1. In some embodiments, the calcium phosphate coating may be hydroxyapatite (HAP) with a Ca / P = 1.5 to 1.67, tricalcium phosphate (TCP) with a Ca / P = 1.5, tetracalcium phosphate (TTCP) with a Ca / P = 2.0 and octacalcium phosphate (OCP) with a Ca / P = 1.33. In some embodiments calcium-deficient hydroxyapatite has a Ca / P ratio between 1.67 and 1.5.

[0051] Nanometer scale amorphous and crystalline calcium phosphate coatings may include monocalcium phosphate monohydrate, monocalcium phosphate anhydrous, dicalcium phosphate dehydrate, dicalcium anhydrous, octacalcium phosphate, apatite, hydroxyapatite, tricalcium phosphate, heptacalcium phosphate, pentacalcium phosphate, tetracalcium phosphate, calcium pyrophosphate, calcium metaphosphate, calcium phosphinate, calcium hydroxide phosphate and combinations thereof. In some embodiments, one or more form of calcium phosphate is electrodeposited onto the surface of an electrically conductive substrate as a highly uniform nanometer thick amorphous calcium phosphate coating which is then converted with a basic solution to a crystalline calcium phosphate with a single layer of highly uniform crystals with a vertical orientation. Nanometer scale crystalline calcium phosphate may be created from any ofDocket No. 2891.098AWO the forms or ratios of calcium phosphate described herein by conversion with a basic or alkaline solution.

[0052] The disclosed nanometer thick calcium phosphate coatings have many benefits including increased adhesion strength to the substrate. The increased adhesion strength is related to the nanosecond pulse electrodeposition of nanometer thick amorphous calcium phosphate. The initial nanometer thick amorphous calcium phosphate is adhered to the conductive substrate through ionic interactions making it tightly bound to the surface. The conversion of the nanometer thick amorphous calcium phosphate to nanometer thick crystalline calcium phosphate creates nanometer crystals with a vertical orientation that is still ionically bound to the surface. Both amorphous, crystalline and semicrystalline nanometer thick calcium phosphate coating have strong ionic bonding to the surface. The increased bonding strength to the surface translates in a very high resistance of the coating to delaminate from the substrate. The nanometer size calcium phosphate coating can expand and contract with the surface of the substrate. The nanometer size calcium phosphate coating demonstrates far less brittleness than the micron size coatings. There also is less stress introduced into the nanometer size calcium phosphate coating during bending of the substrate. In some embodiments, the disclosed nanometer thick calcium phosphate coatings are more resistant to bending, twisting, and tensile forces than micron thick calcium phosphate coatings.

[0053] The thickness of the electrodeposited nanometer thick amorphous calcium phosphate is nanometers thick. The thickness of the crystalline calcium phosphate coating may be approximately the same as the initial amorphous calcium phosphate coating as the conversion process does not add additional calcium phosphate but changes the morphology. When describing nanometer thick amorphous, crystalline and semicrystalline calcium phosphate the term calcium phosphate may be used as a way to describe each form of calcium phosphate. The thickness of the calcium phosphate coating on the substrate may be about 1 nm, about 5 nanometers, about 10 nanometers, about 15 nanometers, about 20 nanometers, about 25 nanometers, about 30 nanometers, about 35 nanometers, about 40 nanometers, about 45 nanometers, about 50 nanometers, about 55 nanometers, about 60 nanometers, about 65 nanometers, about 70 nanometers, about 75 nanometers, about 80 nanometers, about 85 nanometers, about 90 nanometers, about 95 nanometers, about 100 nanometers, about 105Docket No. 2891.098AWO nanometers, about 110 nanometers, about 115 nanometers, about 120 nanometers, about 125 nanometers, about 130 nanometers, about 135 nanometers, about 140 nanometers, about 145 nanometers, about 150 nanometers, about 200 nanometers, about 250 nanometers, about 300 nanometers, about 350 nanometers, about 400 nanometers, about 450 nanometers, about 500 nanometers, about 550 nanometers, about 600 nanometers, about 650 nanometers, about 700 nanometers, about 750 nanometers, about 800 nanometers, about 850 nanometers, about 900 nanometers, about 950 nanometers, or about 1000 nanometers.

[0054] Micron or greater thickness of the calcium phosphate coating is substantial enough to act independently from the base material leading to delamination of the calcium phosphate coating from the surface. Delamination of coatings may lead to unpredictable delivery of drug or compounds in the coating that were designed to be slow released. The micron thickness of plasma spray coatings may exceed machine tolerances of tight fitting parts or pass throughs parts. In some embodiments, micron thick coating cannot be used on precision parts or parts with precise dimensions.

[0055] In some embodiments, the disclosed nanometer thick calcium phosphate coating is a synthetic calcium phosphate coating created using nanosecond pulse electrodeposition. Electrodeposition may be carried out using a power supply connected to electrodes in a conductive liquid, electrolyte or aqueous solution. The electrodeposition system includes an anode and cathode, where the substrate to be coated is electrically connected to either the anode or the cathode. The electrodeposition substrate may act as either the anode or cathode. The counter electrode, the electrode not electrically connected to the substrate, is electrically connected to the deposition liquid. In some embodiments, a power supply is connected to a pulse generator such that nanosecond electric pulses are provided to electrodes which are electrically connected to the substrate and the liquid and the charged substrate attracts material of the opposite charge in the liquid which is electrodeposited on the surface. The liquid may be an aqueous liquid or an ionic polar liquid. Any liquid suitable for conducting electrical pulses may be used and may be optimized. Electrodeposition is discussed in Li, TT., Ling, L., Lin, MC. et al. Recent advances in multifunctional hydroxyapatite coating by electrochemical deposition. J Mater Sci 55, 6352–6374 (2020) and Drevet R, Benhayoune H. Electrodeposition of Calcium Phosphate Coatings on Metallic Substrates for Bone Implant Applications: A Review. Coatings.Docket No. 2891.098AWO 2022; 12(4):539, both of which are hereby incorporated by reference in their entirety. Electrodeposition is discussed in related application PCT / US24 / 48303 published as WO 2025 / 072258 and hereby incorporated by reference in its entirety.

[0056] The duration and amplitude of the pulses used to deposit nanometer thick amorphous calcium phosphate onto the substrate surface are in a range of 1 to 200,000 volts with a duration of 10 to 1,000 nanoseconds with a period of 0.001 to 10,000 hertz. Some embodiments utilize voltage in the range of 100 to 200,000 volts and may be called high voltage embodiments as the voltages used are high compared to traditional electrodeposition. Adjusting the parameters of the voltage, amperage, pulse duration, deposition liquid concentration, pulse frequency and total electrodeposition pulse number allow for control over the electrodeposition of material onto a substrate including the thickness, nanoscale morphology, nanoparticle size, the number of nucleation sites, deposition geometry and combinations thereof.

[0057] The electrodeposition of amorphous calcium phosphate on conductive substrates at the nanoscale allows for precise uniformity of coating. The uniformity in coating of a material on a substate may be the uniformity in coverage of the substrate with the electrodeposition material. The uniformity in coating may be in the strength of adhesion of the material to the substrate. The uniformity in the coating may be in the thickness of the calcium phosphate on the substrate. The uniformity may be in the nanoscale morphology of the calcium phosphate on the substrate. In some embodiments, the uniformity of electrodeposition of amorphous calcium phosphate onto a conductive substrate, of even highly complex geometries, allows for the creation of highly uniform single layer of nanometer size calcium phosphate crystals. The uniformity of the crystalline calcium phosphate coating may be in the uniformity of the crystal size, crystal spacing, crystal vertical orientation, crystal density and combinations thereof. The uniformity may be in one or more properties of the single layer of nanometer sized crystals coating the substrate. The uniformity in the coating may be any combination of thickness, nanoscale morphology, adhesive strength and percent of substrate coated. The electrodeposition of material may be less than a nanometer thick per pulse of electricity. The electrodeposition of material may be on the molecular scale with the electrodeposition of single molecules on the substrate per electric pulse.Docket No. 2891.098AWO

[0058] In some embodiments nanosecond electrochemical deposition or electrodeposition is used to coat the conductive substrate in a nanometer thick coating of calcium phosphate. In some embodiments, nanosecond electrodeposition creates a nanometer thick coating of amorphous calcium phosphate on the surface of the conductive substrate. The voltage used in electrodeposition may influence the size and strength of the nanoparticles of calcium phosphate that coats the substrate. The voltage used for nanosecond pulse electrodeposition used to create nanometer calcium phosphate coatings may be about 1 volt, about 5 volts, about 10 volts, about 15 volts, about 20 volts, about 30 volts, about 40 volts, about 50 volts, about 60 volts, about 70 volts, about 80 volts, about 90 volts, about 100 volts, about 200 volts, about 300 volts, about 400 volts, about 500 volts, about 600 volts, about 700 volts, about 800 volts, about 900 volts, about 1,000 volts, about 2,000 volts, about 3,000 volts, about 4,000 volts, about 5,000 volts, about 6,000 volts, about 7,000 volts, about 8,000 volts, about 9,000 volts, about 10,000 volts, about 20,000 volts, about 30,000 volts, about 40,000 volts, about 50,000 volts, about 100,000 volts, about 150,000 volts or about 200,000 volts.

[0059] The frequency of the nanosecond electric pulses used in electrodeposition to create nanometer thick calcium phosphate coatings may be measured in cycles per second referred to as hertz. The frequency of the electric pulses may be or about 0.001 hertz, or about 0.005 hertz, or about 0.01 hertz, or about 0.015 hertz, or about 0.02 hertz, or about 0.025 hertz, or about 0.03 hertz, or about 0.035 hertz, or about 0.04 hertz, or about 0.045 hertz, or about 0.05 hertz, or about 0.055 hertz, or about 0.06 hertz, or about 0.065 hertz, or about 0.07 hertz, or about 0.075 hertz, or about 0.08 hertz, or about 0.085 hertz, or about 0.09 hertz, or about 0.095 hertz, or about 0.1 hertz, or about 0.5 hertz, or about 1 hertz, or about 5 hertz, or about 10 hertz, or about 20 hertz, or about 50 hertz, or about 75 hertz, or about 100 hertz, or about 250 hertz, or about 500 hertz, or about 750 hertz, or about 1000 hertz, or about 5000 hertz or about 10,000 hertz.

[0060] The electric pulses used for the electrodeposition of nanometer thick calcium phosphate coatings may have a duration of about 1 nanosecond, or about 5 nanoseconds, or about 10 nanoseconds, or about 15 nanoseconds, or about 20 nanoseconds, or about 25 nanoseconds, or about 30 nanoseconds, or about 35 nanoseconds, or about 40 nanoseconds, or about 45 nanoseconds, or about 50 nanoseconds, or about 55 nanoseconds, or about 60 nanoseconds, or about 65 nanoseconds, or about 70 nanoseconds, or about 75 nanoseconds, or about 80Docket No. 2891.098AWO nanoseconds, or about 85 nanoseconds, or about 90 nanoseconds, or about 95 nanoseconds, or about 100 nanoseconds, or about 105 nanoseconds, or about 110 nanoseconds, or about 115 nanoseconds, or about 120 nanoseconds, or about 125 nanoseconds, or about 130 nanoseconds, or about 135 nanoseconds, or about 140 nanoseconds, or about 145 nanoseconds, or about 150 nanoseconds, or about 200 nanoseconds, or about 250 nanoseconds, or about 300 nanoseconds, or about 350 nanoseconds, or about 400 nanoseconds, or about 450 nanoseconds, or about 500 nanoseconds, or about 550 nanoseconds, or about 600 nanoseconds, or about 650 nanoseconds, or about 700 nanoseconds, or about 750 nanoseconds, or about 800 nanoseconds, or about 850 nanoseconds, or about 900 nanoseconds, or about 950 nanoseconds, or about 1000 nanoseconds.

[0061] The thickness of one or more calcium phosphate coating on the substrate may be about 1 nanometer, or about 2 nanometers, or about 5 nanometers, or about 10 nanometers, or about 15 nanometers, or about 20 nanometers, or about 25 nanometers, or about 30 nanometers, or about 35 nanometers, or about 40 nanometers, or about 45 nanometers, or about 50 nanometers, or about 55 nanometers, or about 60 nanometers, or about 65 nanometers, or about 70 nanometers, or about 75 nanometers, or about 80 nanometers, or about 85 nanometers, or about 90 nanometers, or about 95 nanometers, or about 100 nanometers, or about 105 nanometers, or about 110 nanometers, or about 115 nanometers, or about 120 nanometers, or about 125 nanometers, or about 130 nanometers, or about 135 nanometers, or about 140 nanometers, or about 145 nanometers, or about 150 nanometers, or about 200 nanometers, or about 250 nanometers, or about 300 nanometers, or about 350 nanometers, or about 400 nanometers, or about 450 nanometers, or about 500 nanometers, or about 550 nanometers, or about 600 nanometers, or about 650 nanometers, or about 700 nanometers, or about 750 nanometers, or about 800 nanometers, or about 850 nanometers, or about 900 nanometers, or about 950 nanometers, or about 1000 nanometers.

[0062] The calcium phosphate coating may be electrodeposited as calcium phosphate nanoparticles onto the conductive substrate which may form a uniform coating nanometers thick. The size of the nanoparticles of calcium phosphate that are electrodeposited on the conductive substrate may depend on the voltage, frequency, pulse duration, electrolyte concentration and combinations thereof. The size of the nanoparticles of calcium phosphate electrodeposited on the substrate may be about 0.1 nm, or about 0.25 nm, or about 0.5 nm, or about 0.75 nm, or about 1Docket No. 2891.098AWO nm, or about 5 nanometers, or about 10 nanometers, or about 15 nanometers, or about 20 nanometers, or about 25 nanometers, or about 30 nanometers, or about 35 nanometers, or about 40 nanometers, or about 45 nanometers, or about 50 nanometers, or about 55 nanometers, or about 60 nanometers, or about 65 nanometers, or about 70 nanometers, or about 75 nanometers, or about 80 nanometers, or about 85 nanometers, or about 90 nanometers, or about 95 nanometers, or about 100 nanometers, or about 105 nanometers, or about 110 nanometers, or about 115 nanometers, or about 120 nanometers, or about 125 nanometers, or about 130 nanometers, or about 135 nanometers, or about 140 nanometers, or about 145 nanometers, or about 150 nanometers, or about 200 nanometers, or about 250 nanometers, or about 300 nanometers, or about 350 nanometers, or about 400 nanometers, or about 450 nanometers, or about 500 nanometers, or about 550 nanometers, or about 600 nanometers, or about 650 nanometers, or about 700 nanometers, or about 750 nanometers, or about 800 nanometers, or about 850 nanometers, or about 900 nanometers, or about 950 nanometers, or about 1000 nanometers.

[0063] The disclosed calcium phosphate coatings may be applied to conductive surfaces using nanosecond electrodeposition at temperatures that do not damage or alter the properties of metallic substrates. In some embodiments, the nanometer calcium phosphate coatings are deposited at temperatures that do not affect metallic implants or properties of the implants in contrast to other current techniques to create micron thick calcium phosphate coatings. Current techniques used to create micron thick calcium phosphate coating on metal implants includes plasma spray, RF-magnetron sputtering and hydrothermal thermal methods. Plasma spraying was a common commercial technique to apply micron thick calcium phosphate coatings to implants. Plasma spray HAP coating involves heating the HAP material to a semi-molten or molten state and propelling it on a metal substrate. The plasma flame temperature has been reported to be in the range of 6,000 degrees centigrade to 16,000 degrees centigrade. These temperatures may damage one or more material or material property in a medical implant and prevent co- deposition of heat sensitive dopants. Portions of metal implants or metal implants with low thermal mass may suffer metallic phase changes at these temperatures thereby jeopardizing the physical properties of the implant. The plasma spray is at a high temperature that can heat parts to the extent of altering the temper of the base metal, especially in parts with small thermal masses such as wire, screws and plates. Plasma spray heating may alter mechanical strength,Docket No. 2891.098AWO electrical conductivity, magnetic properties and corrosion resistance of the metal substrate. The disclosed coatings may be applied at temperatures far lower than required to temper commonly used metals or metal alloys used in medical applications thus physical properties, such as hardness, of the metals or alloys are not changed during the coating process as is possible with plasma spray coating. Thus, the process of applying the calcium phosphate coating will not damage the temper, corrosion resistance, magnetic properties or tensile strength of an implant as can happen with plasma sprayed calcium phosphate surfaces.

[0064] Nanosecond electrodeposition of calcium phosphate may occur at temperatures that do not inactive dopants. Nanosecond electrodeposition of a calcium phosphate coating may be applied to a conductive substrate at about -30 degrees centigrade, at about -25 degrees centigrade, at about -20 degrees centigrade, at about -15 degrees centigrade, at about -10 degrees centigrade, at about -5 degrees centigrade, at about 0 degrees centigrade, at about 5 degrees centigrade, at about 10 degrees centigrade, at about 15 degrees centigrade, at about 20 degrees centigrade, at about 25 degrees centigrade, at about 30 degrees centigrade, at about 35 degrees centigrade, at about 40 degrees centigrade, at about 45 degrees centigrade, at about 50 degrees centigrade, at about 55 degrees centigrade, at about 60 degrees centigrade, at about 65 degrees centigrade, at about 70 degrees centigrade, at about 75 degrees centigrade, at about 80 degrees centigrade, at about 85 degrees centigrade, at about 90 degrees centigrade, at about 95 degrees centigrade, at about 100 degrees centigrade, at about 105 degrees centigrade, at about 110 degrees centigrade, at about 115 degrees centigrade, at about 120 degrees centigrade, at about 130 degrees centigrade or at one or more of the foregoing temperatures.

[0065] Methods of depositing calcium phosphate coatings such as continuous direct current electrodeposition have inconsistent uniformity of micron thick calcium phosphate coating which is often a result of gas bubble formation on the surface of the conductive substrate. Continuous direct current electrodeposition suffers from uneven deposition of coating materials and areas with larger surface area are coated at different micron thickness than area of smaller surface area such as fine edges and points. The resulting uneven micron thick coating results in areas of the coated substrate with different properties than other areas with more coating. The problem is particularly evident in substrates with complex geometries which may suffer discontinuous, uneven micron thick coatings. In an embodiment, an uneven calcium phosphate micron thickDocket No. 2891.098AWO coating with antimicrobial additives may allow biofilm attachment in areas where the coating is uneven due to lower concentrations of antimicrobial agents. Areas of less coating thickness may wear or dissolve faster than areas of thicker coating resulting in a different cellular response to the surface coating in worn areas. A nanometer thick uniform calcium phosphate coating has many advantages including predictable life span, uniform strength and consistent host cellular interactions with the surface.

[0066] The disclosed nanometer thick calcium phosphate coatings have a crystal vertical orientation unlike other commercially available nano scale thickness calcium phosphate crystals that are dip coated onto the substrate from a solution of precipitated calcium phosphate crystals which can collect unevenly on the surface. The dip coating method also relies on the van der waals forces for adhesion of the nanoscale crystals. The larger the crystals the more susceptible the crystals are to coming off. This weak adhesion also makes the surface very susceptible to damage if it gets wet during storage. The crystals in dip coating are also randomly oriented on the surface with a slight bias to lying flat on the surface which may not be an optimal orientation to resist the adhesion of bacteria and provides few bonding atoms to the environment.

[0067] It is believed micron thick calcium phosphate coatings bond primarily with van der Waals force and are formed from micron size particles adhering to the substrate in a variety of ways. In some instances, hydrophobic interactions are used to force calcium phosphate to a surface. Unlike ionic or covalent bonds, these attractions do not result from a chemical electronic bond and are comparatively weak and therefore more susceptible to breaking. The van der Waals force quickly vanishes at longer distances between interacting molecules. This may explain why micron thick calcium phosphate coatings delaminate. The further the calcium phosphate coating material is from the substrate the less likely it is bonding with the substrate and more likely it is bonding with the coating.

[0068] Without being limited to any particular theory, the nanometer thick calcium phosphate coating appears to nucleate as very small nanoparticles that directly adhere to the substrate through ionic bonds. Each nanoparticle of the calcium phosphate is bound to the conductive substrate through ionic charge attraction including each additional nanoparticle as the calcium phosphate coating grows in thickness.Docket No. 2891.098AWO

[0069] The disclosed ionic bonded nanometer thick calcium phosphate coating adheres to the substrate with more adhesion force, and demonstrate superior resistance to delamination, lack of brittleness, improved tensile strength and compressive resistance than micron thick calcium phosphate coatings. In some embodiments, nanometer thick calcium phosphate coatings are created by nanosecond electric pulse electrodeposition. In some embodiments, nanosecond electric pulse created nanometer calcium phosphate coatings are significantly more resistant to delamination. In some embodiments, electrodeposited nanometer thick calcium phosphate coating is more resistant to delamination than other methods of depositing calcium phosphate coatings, such as plasma spray coating. The nanosecond electric pulsed electrodeposition of the nanometer thick amorphous calcium phosphate is very well adhered to the surface due to the ionic bonds formed with the surface due to the ionization of the surface during the direct current electric pulses. Individual crystals are grown directly attached to the surface, making the surface much more durable, less prone to delamination or sloughing in solution. The more durable crystalline coating can withstand autoclaving in addition to eBeam, gamma, UV and peroxide vapor sterilization and mechanical stress. In some embodiments, nanometer thick amorphous calcium phosphate coating of conductive substrate is converted to delamination resistant nanometer thick crystalline calcium phosphate using a base while maintaining the original thickness of the nanometer thick amorphous calcium phosphate coating.

[0070] Calcium phosphate coatings are used to improve the biological compatibility of medical implants. Medical implants utilize metals and alloys of stainless steel (SS), titanium (Ti), cobalt (Co), nickel-titanium (NiTi), magnesium (Mg), cobalt-chromium (CoCr) and others. These metals and alloys have physical properties, such as high strength, high fracture toughness, hardness, corrosion resistance and biocompatibility that make them useful for metallic bone implants. Calcium phosphate coatings improve how the cells interact with the implant and can lead to faster healing and stronger implant fixation. Calcium phosphate, in the form of hydroxy appetite (HAP), is the main inorganic component present in bone and teeth and may be used as coating on medical implants. HAP coatings lead to osteointegration of the implant by creating a direct structural and functional connection between ordered living bone and the surface of a load- carrying implant. The HAP surface induces osteoblasts to form over the surface of the implant improving bone fixation. Though HAP is beneficial on the surface of medical implants for improving implant integration into the recipient commonly used micron thick coatings sufferDocket No. 2891.098AWO from a number of problems. Commercial micron thick HAP used in implants suffer from brittleness, low tensile strength and low fracture resistance. The disclosed nanometer thick calcium phosphate coatings have properties that are novel as compared to the same coatings in the micron scale of thickness which are currently used in commercial calcium phosphate coating of metal implants.

[0071] Some of the benefits of the disclosed uniform nanometer thick coating include nanometer crystals are the many unsaturated bonds for binding biomolecules such as DNA, RNA, proteins, peptides, hormones, antibodies and antibody drug conjugates. The nanometer crystals may also bind molecules such as ions, small molecules, receptor ligands, agonists and antagonists. The disclosed nanometer thick calcium phosphate can be used as a carrier for various bioactive elements or molecules such as growth factors, antivirals, antibiotics, and antimicrobials. Additionally, the uniform nanometer thick calcium phosphate coating can be tuned to create predetermined nanometer thickness and predetermined nanometer size crystals. By controlling the thickness of the calcium phosphate coating and the size of the crystals it is possible to fine tune the calcium phosphate coating to bind specific molecules to the coating. Specific calcium phosphate coating can be engineered to bind to specific biological or chemical molecules to influence specific biologic functions such as promoting bone growth, promoting bone strength, creating antimicrobial properties, and creating a coating with a desired decay velocity. In some embodiments, a nanometer thick crystalline calcium phosphate coating with a plurality of calcium phosphate crystal about 1 nanometer will optimally bind a desired compound. In some embodiments, larger nanometer calcium phosphates crystals may be used to coat a metal implant in tissues with larger cellular structures. In some embodiments, smaller nanometer calcium phosphates crystals may be used to coat a metal implant in tissues with smaller cellular structures. In some embodiments, tuning crystal size to tissue type improves cellular integration of a device such as an implant with a nanometer calcium phosphate surface.

[0072] In some embodiments, a plurality of calcium phosphate crystals are about or 1, or about 2, or about 3, or about 4, or about 5, or about 6, or about 7, or about 8, or about 9, or about 10, or about 12, or about 15, or about 20, or about 25, or about 30, or about 35, or about 40, or about 45, or about 50, or about 55, or about 60, or about 65, or about 70, or about 75, or about 80, or about 85, or about 90, or about 95, or about 100, or about 105, or about 110, or about 115, orDocket No. 2891.098AWO about 120, or about 125, or about 130, or about 135, or about 140, or about 145, or about 150, or about 155, or about 160, or about 165, or about 170, or about 175, or about 180, or about 185, or about 190, or about 195, or about 200, or about 225, or about 250, or about 275, or about 300, or about 325, or about 350, or about 375, or about 400, or about 425, or about 450, or about 475, or about 500, or about 525, or about 550, or about 575, or about 600, or about 625, or about 650, or about 675, or about 700, or about 725, or about 750, or about 775, or about 800, or about 825, or about 850, or about 875, or about 900, or about 925, or about 950, or about 975, or about 1000 nanometers in height, width, length or combinations thereof. In some embodiments, the aspect ratio of the crystal may be used to determine one or more dimension.

[0073] The calcium phosphate coating can be composed of quantifiable nanoscale structures such as crystals. Nanoscale amorphous spheres or balls may be electrodeposited onto the surface of the substrate. Nanometer size spheres may be electrodeposited to form a uniform coating or non-uniform coating. The coating can utilize and preserve pre-existing nanoscale features on a substrate which could be useful for medical devices and implants and may be configured with various additional biomolecules and achieve an FDA nanotechnology designation.

[0074] The disclosed uniform nanometer thick calcium phosphate coatings may be used to release dopants at a specific rate based on the rate of dissolution of the calcium phosphate coating. The rate of dissolution may be referred to as rate of decay of the coating. The nanometer thick calcium phosphate coating may dissolve at a pH dependent rate. The coating may dissolve at a rate dependent on the local environment. In some embodiments, calcium phosphate coatings may include a homogenous concentration of dopants. In some embodiments, calcium phosphate coatings may include a heterogeneous concentration of dopants. In some embodiments, dopants may be concentrated near the substrate. In some embodiments, dopants may be concentrated at the very surface of the coating in contact with the environment. In some embodiments, calcium phosphate coatings may include a gradient of dopants. A gradient of dopant may be created by soaking a calcium phosphate coating into a solution, such as zinc, where the dopant is substituted for calcium at decreasing concentrations with increasing depth into the coating. In an embodiment, a zinc gradient is formed by soaking the nanometer thick calcium phosphate crystalline surface in a solution of zinc where the zinc substitutes for calcium and the highest concentration of zinc is at the surface of the coating and the concentration of zinc decreases withDocket No. 2891.098AWO increasing depth into the coating. The calcium phosphate coating may include one or more layers of nanometer thick calcium phosphate coatings. In some embodiments, nanosecond electrodeposition is performed to create one or more layer of nanometer thick calcium phosphate coatings such that each layer may be defined by the inclusion of one or more dopant, one or more nanoparticles, one or more calcium to phosphate ratio or combinations thereof. One or more nanometer thick calcium phosphate layer may include one or more dopant. One or more dopants may be present at different nanometer thickness of the coating as measured from the substrate or the surface. In some embodiments, specific dopants are included at specific thickness of the coating such that the outermost layer, in contact with the environment, and thereby releases a first dopant for a period of time and then a layer below may release a second dopant for a period of time and a layer below may release a third dopant, a layer below may release a fourth dopant, a layer below may release a fifth dopant, a layer below may release sixth dopant, a layer below may release a seventh dopant and so on for any desired amount of dopants. Dopants may be slow release or release in bursts. In some embodiments, a slow release dopant may be a low concentration dopant homogenized in the calcium phosphate coating. In some embodiments, a dopant is released in burst when it is highly concentrated to a specific layer of the calcium phosphate coating. In some embodiments a dopant may be between one or more calcium phosphate layers, which may be termed interstitial to one or more layers.

[0075] The cellular environment may affect the rate of decay. Nanometer thick calcium phosphate coatings in contact with immune cells, cancer cells or osteoclasts may require more chemical resistance or specific dopants to preserve the coating for a predetermined time period. Medical implants that are in contact with areas of bone, cartilage and cellular healing may benefit from dopants selected for this environment. The coating may include dopants such as metal ions known to have antimicrobial or antibacterial effects, such as silver, copper, or zinc that create an antimicrobial effect for a predetermined length of time. In some embodiments, a biological effect may last for at about one month, about two months, about three months, about four months, about five months, about six months, about seven months, about eight months, about nine months or about twelve months. In some embodiments, a biological effect may last at about one year, about two years, about three years, about four years or about five years. In some embodiments, a biological effect may be about a week, about two weeks, about three weeks or about four weeks. A biological effect may include promoting or inhibiting osteoblasts,Docket No. 2891.098AWO osteocytes, osteoclasts and bone lining cells, stem cells, cellular receptors and cellular pathways. A biological effect may include inhibiting microorganisms such as bacteria, viruses, parasites, fungi and protozoa. A biological effect may include bioresorption, cell adhesion activity, cell proliferation activity, cell migration activity, cell differentiation activity, anti-bacterial activity, bactericidal activity, anti-inflammatory activity, osseointegration activity, biocorrosion activity, cell differentiation activity, immuno-modulating activity during acute or chronic inflammation or any combination of these. Biological effects may include the nanometer calcium phosphate crystal size of the coating thereby encouraging mesenchymal stem cells to differentiate into osteoblast. Additional biological effects include osteoblasts excreting extracellular matrix to form bone on growth. Biological effects include osteointegration of a coated implant. In some embodiments, a nanometer thick crystalline calcium phosphate coating includes dopants to promote osteointegration.

[0076] In some embodiments the nanometer thick calcium phosphate coating promotes mesenchymal stem cells to differentiate into osteoblasts. The osteoblast may then excrete extracellular matrix to form bone growth. In some embodiments the nanometer thick calcium phosphate coating nanometer crystal surface promotes mesenchymal stem cells to differentiate into osteoblasts wherein they form bone growth on the surface. The rapid promotion of new bone growth on the disclosed surface may allow for the rapid fixation of medical devices, metal hardware and combinations thereof.

[0077] The nanometer size calcium phosphate coating also prevents over osteointegration that occurs in micron thick calcium phosphate coatings. In micrometer thick calcium phosphate coatings, such as HAP, osteoblasts are able to integrate into the micron sized features of the coating. Since cells can adhere and grow in the micron size features it is possible for osteoclastic pits to form in the micron thick coating and subsequent bone formation in the micron size pits. The micron thick calcium phosphate coating can also lead to difficulty of removing the implant after osteoclast activity burrows pits into the coating forming bone in the new pits. The nanometer size calcium phosphate coating doesn’t have enough thickness for a single cell to fit into a surface coating cavity. In an embodiment, nanometer size coatings do not have large enough pits created by osteoclast resorption to allow cell growth inside a coating feature. Nanometer size coatings require cells to attach to the coating and to biological material, such asDocket No. 2891.098AWO bone or cartilage, increasing speed and strength of implant osteointegration and resistance to delamination of the coating.

[0078] The disclosed nanometer thick calcium phosphate coating can be used to trigger specific phases of bone development. In some embodiments the coating will release dopants after certain periods of time to effect one or more bone phase. Bone remodeling is a dynamic process composed of four phases: activation, resorption, reverse, and formation. The first phase, Activation, is initiated when bone lining cells are activated and increase the surface expression of receptor activator for nuclear factor κ B ligand (RANKL). The second phase, Resorption, is initiated with the activation of counterpart RANK, which is expressed on pre-osteoclasts and triggers osteoclast differentiation. Osteoclasts proceed with bone resorption and signaling to initiate osteoid matrix formation by osteoblasts. The third phase, Reversal, begins when new bone formation by osteoblasts overtakes bone resorption by osteoclasts. The last phase, Formation, completes the remodeling process, in which a new bone matrix is deposited by osteoblasts, mineralizing the collagenous osteoid matrix. Any one or more of these phases can be promoted or inhibited with dopants in the disclosed nanometer scale calcium phosphate coating. One or more nanometer thick calcium phosphate coating may act to promote or inhibit one or more phase of bone remodeling. One or more calcium phosphate coating in combination with one or more dopant, nanoparticle or combinations thereof may act to promote or inhibit one or more phases of activation, resorption, reverse and formation. In some embodiments, a nanocrystal calcium phosphate coating may contain dopants such as RANKL to promote bone activation. The relative concentration of Receptor activator of NF-κB ligand (RANKL) and osteoprotegerin (OPG) in bone is a major determinant of bone mass and strength. In some embodiments, OPG and RANKL are dopants in the nanometer thick calcium phosphate coating to promote bone mass and strength.

[0079] Calcium phosphate coating may be used to coat conductive substrates such as indium tin oxide (ITO) glass. ITO glass is conductive and would be a suitable substrate for nanometer size calcium phosphate coating. In some embodiments, ITO glass coated with calcium phosphate could be used in microwell plates, slides, petri dishes, and other labware. In some embodiments, microplates with ITO glass coated with nanometer thick calcium phosphate coating, wherein wells of a microplate are coated with calcium phosphate which binds nucleic acid and when cellsDocket No. 2891.098AWO are place in the wells the cell contacts the bound nucleic acid on the cell surface, internalizes the nucleic acid in the process of transfection. In some embodiments, nanometer thick calcium phosphate coatings may transfect one or more cells or tissues. One transfection technique is to use calcium phosphate as a solution to mix with a solution of nucleic acid to transfect cells, where the calcium phosphate assists in the precipitation of the nucleic acid onto the surface of the cell for transfection to occur. Calcium phosphate coatings may allow transfection to occur through glass surfaces such as microplates and slides or using nanoparticles with calcium phosphate coatings.

[0080] Nanometer thick calcium phosphate coatings may be on any conductive substrate including metals and alloys. In some embodiments, non-conductive substrates may be treated to have a conductive surface and then be suitable for electrodeposition of calcium phosphate. In some embodiments, a medical device or implant may have at least one conductive surface that may be coated with nanometer thick calcium phosphate. Conductive substrates include metals, alloys or any combination thereof, Conductive substrates include metal and alloy implants. Metal and alloy implants may include medical devices, screws, stents, mesh, plates, pins, wire, nails, rods, tubes, balls, joints and combinations thereof.

[0081] The topology of a conductive substrate may be preserved or engineered. If a conductive substrate has a nanometer scale topology already present a uniform nanometer thick coating may be applied and preserve the existing nanometer scale topology. The ability of nanometer size calcium phosphate coatings to preserve nanometer coatings allows highly functionalized surfaces with the benefits of nanotechnology features to also include the benefits of an additional nanotechnology in the coating itself. Many coating technologies cannot preserve nanometer size features. In some embodiments, nanotubes are present on a surface and a nanometer size calcium phosphate coatings preserves the nanotubes. In some embodiments, nanoparticles are present and a nanometer size calcium phosphate coating preserves the topology of the nanoparticles. In some embodiments, if a substrate lacks a nanometer scale topology one can be created by varying the nanometer size thickness of the coating or the nanometer size of the crystals. Creating a nanometer scale surface on a substrate greatly increases the surface area for chemical or biological reactions. Nanometer size crystal surfaces may impart new functionality to a surface such as different binding, optical, and electric properties. A conductive surface mayDocket No. 2891.098AWO include a micrometer scale topology that may be preserved and enhanced with nanometer scale features such as nanometer thick calcium phosphate coatings, calcium phosphate crystals or combinations thereof. A surface topology may be engineered using surfacing techniques, the additional of nanoparticles, nanotubes, microparticles, microtubes, anodization, passivation, etching, or any combination thereof.

[0082] In an embodiment, a surface, such as a conductive surface, already has an engineered nanometer scale topology but is made from a base material that has a non-desired or inert biological interaction. A coating of 1-50 nanometers of calcium phosphate or doped calcium phosphate can be added preserving the topology but adding the enhanced performance of the bioactive surface chemistry. In an embodiment, nanotubes are coated with 5-10 nanometers of Zinc doped calcium phosphate whereby the nanotube topology is preserved but gains improved macrophage, endothelial cell, mesenchymal stem cell and osteoblast function in addition to antimicrobial properties from the coating.

[0083] In some embodiments, a conductive surface does not have a nanoscale topology already present and a nanoscale topology can be engineered on the surface with a nanometer thick calcium phosphate layer and a post processing crystalline conversion step resulting in tunable size and density of the crystalline calcium phosphate surface.

[0084] The base layer of amorphous calcium phosphate produced with the nanosecond electric pulsed electrodeposition method has uniformity over the entire surface including inside more porous geometries. The pulsed electrodeposition method is also referred to as the pulsed electrochemical deposition method. The crystalline conversion also can then convert the uniform base layer into the nano-crystalline surface. The converting solution does not add any material to the surface, just converts what is there. Surface uniformity and thickness is preserved based on the initial amorphous coating process. The nanometer thin calcium phosphate coating has reduced risk of delamination which would lead to unpredictable release of a dopant in the coating.

[0085] A basic pH solution may be used to convert amorphous calcium phosphate to crystalline calcium phosphate. In some embodiments, a calcium phosphate coating is soaked in basic solution. A feature of the nanometer thick amorphous calcium phosphate coating or layer being converted to a crystalline calcium phosphate coating or layer is that the crystal size and densityDocket No. 2891.098AWO can change based on the amorphous calcium phosphate layer thickness and the conversion step. In some embodiments, a smaller nanometer scale calcium phosphate crystal size is made using a thinner nanometer thick amorphous calcium phosphate layer followed by conversion with a relatively higher concentration of base that is heated and the conversion reaction occurs for a shorter period of time. In some embodiments, larger nanometer calcium phosphate crystals are made using a relatively thicker nanometer amorphous calcium phosphate coatings that is converted more slowly with a lower concentration base that is at a cooler temperature and reacted for a longer period of time. In some embodiments, the longer the conversion process or conversion step the larger the nanometer calcium phosphate crystals produced. In some embodiments, the shorter the conversion process the smaller the nanometer calcium phosphate crystals. In some embodiments an amorphous calcium phosphate coating is converted with a base, such as sodium hydroxide, resulting in a conductive substrate with a surface of vertically oriented nanometer calcium phosphate crystals. The pH of the conversion solution may be used to adjust the size of the calcium phosphate crystals. For example, a very basic solution such as pH 10 may convert amorphous calcium phosphate to smaller nanometer calcium phosphate crystals. In some embodiments, the crystals are approximately 22.5 degrees vertical relative to the substrate. In some embodiments, the vertical crystals are more effective at decreasing bacterial attachment and may physically disrupt bacteria membranes, leading to contact mediated bacterial cell death. In some embodiments, the pointed crystal surface may be difficult for bacterial attachment. The plurality of nanometer crystals may act to inhibit bacterial colonization of a surface thereby producing an antimicrobial effect. This property may be especially useful for medical implants. It may also be useful for conductive surfaces that would benefit from an antimicrobial effect such as bioreactors that may be used as is known in the art such as for cell culture, antibody culture and fermentation.

[0086] The density of nanometer calcium phosphate crystal may include the number of crystals per square unit of area and a number of grams of calcium phosphate per unit of square area. The density of the crystals may be increased or decreased. In some embodiments, increasing or decreasing crystal density is to achieve a desired binding effect, such as binding capacity, as the crystals have many free atoms available for binding. Crystal density may be controlled by the thickness of the initial electrodeposited amorphous calcium phosphate coating. In some embodiments, increasing the amorphous calcium phosphate coating from 100 to 500 nanometersDocket No. 2891.098AWO increases the density of nanometer size calcium phosphate crystals formed during the conversion of amorphous calcium phosphate into crystalline calcium phosphate using a basic solution.

[0087] The density of nanometer calcium phosphate crystals may be about 5 crystals per square micron, about 25, about 50, about 75, about 100, about 125, about 150, about 175, about 200, about 225, about 250, about 275, about 300, about 325, about 350, about 375, about 400, about 425, about 450, about 475 or about 500 crystals per square micrometer.

[0088] The density of nanometer calcium phosphate may in square meters per gram. The density of nanometer calcium phosphate may be about 300 square meters per gram, about 500 square meters per gram, about 1000 square meters per gram, about 2000 square meters per gram, about 3000 square meters per gram, about 4000 square meters per gram, about 5000 square meters per gram, about 6000 square meters per gram, about 7000 square meters per gram, about 8000 square meters per gram, about 9000 square meters per gram, or about 10,000 square meters per gram. In some embodiments, the density in square meters per gram refers to a crystalline calcium phosphate. In some embodiments, the density in square meters per gram refers to semicrystalline calcium phosphate. In some embodiments, the density in square meters per gram refers to amorphous calcium phosphate.

[0089] An amorphous calcium phosphate coating may be converted using a base or alkaline solution. The temperature of the base may influence the crystal size. In some embodiments the base solution for conversion of amorphous to crystalline calcium phosphate is at about 0 degrees centigrade, at about 5 degrees centigrade, at about 10 degrees centigrade, at about 15 degrees centigrade, at about 20 degrees centigrade, at about 25 degrees centigrade, at about 30 degrees centigrade, at about 35 degrees centigrade, at about 40 degrees centigrade, at about 45 degrees centigrade, at about 50 degrees centigrade, at about 55 degrees centigrade, at about 60 degrees centigrade, at about 65 degrees centigrade, at about 70 degrees centigrade, at about 75 degrees centigrade, at about 80 degrees centigrade, at about 85 degrees centigrade, at about 90 degrees centigrade, at about 95 degrees centigrade or at about 100 degrees centigrade.

[0090] Amorphous calcium phosphate may be converted to a crystalline form of calcium phosphate such as hydroxyapatite. In some embodiments, amorphous calcium phosphate (ACP), octacalcium phosphate (OCP), monetite (CaHPO4), and even brushite, may be converted to crystalline calcium phosphate in alkali conditions to produce a nanometer thick coating withDocket No. 2891.098AWO nanometer size calcium phosphate crystals. In other embodiments, amorphous calcium phosphate of various ratios of calcium to phosphate, as described herein, are converted to crystalline calcium phosphate to create a surface with a single layer of nanometer sized crystals that have a vertical orientation. In some embodiments, calcium deficient hydroxyapatite may be converted to nanometer crystalline calcium phosphate. Nanometer thick amorphous calcium phosphate may be formed into nanometer thick semicrystalline calcium phosphate. Semicrystalline calcium phosphate may include amorphous calcium phosphate that has not been completely converted into a crystalline surface. Nanometer thick semicrystalline calcium phosphate may include coatings both amorphous and crystalline calcium phosphate. Semicrystalline can also be amorphous with crystals dispersed throughout. Semicrystalline may be a crystalline and amorphous layers created with independent steps.

[0091] A basic or alkali solution may be used to convert nanometer thick amorphous calcium phosphate to nanometer sized crystalline calcium phosphate. In some embodiments, a basic or alkali solution made from sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide, zinc hydroxide, ammonia, guanidine, barium hydroxide, caesium hydroxide, strontium hydroxide,lithium hydroxide, rubidium hydroxide, Sodium amide, Sodium hydride, and combinations thereof may be used. In some embodiments, a weak base may be used to convert amorphous calcium phosphate to crystalline calcium phosphate. In other embodiments, a strong base may be used to convert amorphous calcium phosphate to crystalline calcium phosphate. In some embodiments heat in addition to a basic or alkali solution is used to convert nanometer thick amorphous calcium phosphate into nanometer thick crystalline calcium phosphate. In some embodiments a combination of a weak base and strong base may be used for conversion of amorphous calcium phosphate to crystalline calcium phosphate. The concentration of the base or alkaline solution may influence the speed of conversion of amorphous calcium phosphate to crystalline calcium phosphate. In some embodiments, the concentration of the base or alkaline solution used is about 0.1mM, about 0.5 mM, about 1 mM, about 2 mM, about 5 mM, about 10 mM, about 20 mM, about 50 mM, about 100 mM, about 250 mM, about 500 mM, about 1 M, about 2 M, about 5 M, or about 10 M.

[0092] The total conversion time may be dependent on the desired density and / or size of nanometer sized crystals. The total conversion time for converting amorphous calcium phosphateDocket No. 2891.098AWO to nanometer sized crystalline calcium phosphate may be about 30 seconds, about 1 minute, about 5 minutes, about 10 minutes, about 15 minutes, about 20 minutes, about 30 minutes, about 45 minutes, about 1 hour, about 1.5 hours, about 2 hours, about 4 hours, about 6 hours, about 8 hours, about 12 hours, about 24 hours, about 36 hours about 48 hours, about 72 hours, about 96 hours, about 125 hours, about 144 hours, about 168 hours, about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks or about 6 weeks.

[0093] In some embodiments, phosphate conversion may involve surface treatment, such as cleaning the conductive surface before the coating process. In some embodiments, a post- treatment, after conversion, such as thermal annealing and additional alkaline treatment to improve the property of coatings and crystals.

[0094] The nanometer thickness also makes it possible for some molecules to be added to the coating by diffusing into the coating. Diffusing molecules into a coating may be done in a liquid or gas.

[0095] Molecules may be added to a nanometer thick calcium phosphate coating by ion substitution or inclusion through the entire coating surface area. Dopants may bond to ions and nanoparticles present in the calcium phosphate coating. In some embodiments, pulsed nanosecond electrodeposition is used to deposit calcium phosphate coatings and to co-deposit dopants. During electrodeposition dopants such as ions may be integrated into the calcium phosphate coating. Dopants may include divalent cations. Divalent cations may be used as dopants in ionic substitutions of calcium phosphate coatings. Some divalent cation dopants used for ionic substitutions include Co2+, Cu2+, Mg2+, Mn2+, Sr2+ and Zn2+. Dopants may include monovalent cations such as K+, Ag+ and Na+ ions. Dopants may include anions such as CO2-, F-. Cl- and SiO4- ions. Dopants may impart biological activity to a conductive surface. Cobalt may promote angiogenesis. Copper may have antibacterial properties. Magnesium may promote bone densification. Manganese may regulate osteoblast differentiation and promote cell adhesion. Strontium may stimulate bone formation and inhibit bone resorption. Zinc may provide antibacterial, anti-inflammatory properties and promote growth of bone. Ionic dopants are discussed in Drevet et al. (2022). Coatings. 12. 539, which is hereby fully incorporated by reference.Docket No. 2891.098AWO

[0096] In some embodiments, the nanometer calcium phosphate vertical crystals have more unbound crystal faces that are free to bind to dopants. In some embodiments, the crystals bind much more protein than a surface without vertical crystals of calcium phosphate. In some embodiments, the vertical crystal surface allows or more protein adsorption leading to more sites for eukaryotic cells attachment. This may be a result of the eukaryotic cells being larger than bacteria or having more fluid cell membranes than bacteria and can conform to the nanometer geometries of the crystalline surface.

[0097] Dopants as described herein may include a variety of drugs, nanoparticles, medicines, antimicrobials, metals, ions and any other dopant as so described herein. Any one or more dopants may be combined with another one or more dopants. Combinations of dopants described herein are embodiments of dopants that may be used with the disclosed nanometer thick calcium phosphate coating. In some embodiments, metals dopants are combined with nanoparticles dopants, anti-inflammatory dopants and pain medicine dopants to create a desired dopant for use with the disclosed calcium phosphate coating. In some embodiments, nanoparticles contain dopants and they are adhered to the substrate wherein the disclosed nanometer thick crystal calcium phosphate coating applied over the nanoparticles and as the nanocrystal calcium phosphate coating dissolves the nanoparticles release dopants. In other embodiments, nanoparticles containing dopants are co-deposited with the calcium phosphate coating which is then converted with a base to a nanometer thick crystalline calcium phosphate coating where the nanoparticles act to release dopants immediately and for a predetermined period of time. In some embodiments, a nanometer thick crystalline calcium phosphate coating contains dopants such as bone growth hormones, proteins, anti-inflammatory medicine, and antimicrobials.

[0098] Dopants may include nanoparticles. Nanoparticles may be any particle of any material that is of nanometer size. In some embodiments, nanoparticles may be in an electrodeposition solution and are co-deposited into the calcium phosphate coating. In some embodiments, pre- existing nanoparticles can be added to a substrate or surface before the nanometer thin calcium phosphate coating. In some embodiments, during the pulsed deposition process the coating will cover and integrate the nanoparticles into the calcium phosphate coating and be physically retained. In some embodiments nanoparticles are adhered to a conductive substrate using adhesive or other chemical bonding that does not affect the conductivity of the substrate suchDocket No. 2891.098AWO that the calcium phosphate is uniformly deposited as nanometer thick coating trapping the nanoparticles. In some embodiments, nanoparticles are added after the formation of the nanometer thick calcium phosphate coating. Additionally, preformed nanoparticles can be incorporated into the calcium phosphate coating. In some embodiments, the nanoparticles are released as the coating dissolves. The nanoparticles may include dopants. The nanoparticles may be metal nanoparticles. Nanoparticles include carbon nanotubes, fullerenes, such as C60 and C70 fullerenes, metal nanoparticles, ceramic nanoparticles, and polymeric nanoparticles. The size of the nanoparticles may influence the optical properties of surfaces or coatings due to the size of the wavelength of light they absorb.

[0099] Dopants may include one or more fluorescent dye, photoluminescent dye, fluorescent protein, radiochemical, marker chemical, reporter molecule or combinations thereof. Marker chemicals and reporter molecules may be used to monitor aspects of the coating. In some embodiments, fluorescent dyes may monitor the dissolution of a coating. In other embodiments, fluorescent dyes may monitor cell growth over the nanometer calcium phosphate coated substrate, such as bone growth, bone fixation, osteoblast growth, osteoclast growth and combinations of cells and cell activity. In some embodiments, using an infrared dye that penetrates tissue, without the need for invasive surgery, will allow monitoring of fixation of medical implants and potential bacterial infections without the risk of introducing new bacteria into the implant. Fluorescent dyes may include, for example, cyanine 3, cyanine 5, Cy7, Cy7.5, rhodamine-based dye molecules, squarylium-based dye molecules, cyanine-based dye molecules, aromatic ring-based dye molecules, oxazine-based dye molecules, carbopyronine-based dye molecules and pyrromethene-based dye molecules. Energy transfer dyes may be used. In some embodiments, fluorescent tagged cells may be identified by energy transfer from dyes present in the calcium phosphate coating.

[0100] A dopant may include one or more therapeutic materials, bioactive molecules, biomaterials, biological agents, biominerals, hydroxyapatite, bioactive calcium phosphate, antibiotics, medicaments, antibacterial agents, anti-viral agents, cariostatics, anti-inflammatory agents, anti-oxidants, organic and mineral fraction of bones and teeth, biologic factors, bioactive substances and cells, drugs, proteins, hormones, enzymes, antigens, antibody, antibody drug conjugate, immunogens, cytotoxins, neurotransmitters, interferons, interleukins, chemokines,Docket No. 2891.098AWO cytokines, extracellular matrix components, ligands and peptides, osteoinductive factors, and combinations thereof.

[0101] Vitamins and minerals are also beneficial to bone growth and remodeling and can be included in calcium phosphate coatings as dopants. A dopant may include one or more of vitamin A (beta-carotene), vitamin B1 (Thiamine HCl), vitamin B2, riboflavin, riboflavin 5′- monophosphate, vitamin B3 (niacin), vitamin B5 (calcium D-pantothenate), vitamin B6, pyridoxine HCl, pyridoxal 5′-phsophate, vitamin B12 (hydroxycobalamin), vitamin C (L- ascorbate), vitamin D3 (cholecalciferol), vitamin E (as mixed tocopherols), vitamin K1 (phylloquinone), vitamin K2 (menatetranone, MK-4), biotin, folate (folic acid), PABA, boron (boron ascorbate), calcium (calcium citrate), chromium, chromium picolinate, chromium ascorbate, copper (copper citrate), magnesium (as magnesium citrate, magnesium malate or magnesium succinate), manganese (manganese ascorbate), molybdenum (molybdenum ascorbate), potassium (potassium citrate), selenium (L-selenomethianone), vanadium (vanadium ascorbate), zinc (zinc picolinate) and combinations thereof. In some embodiments, dopants including vitamins and minerals are slowly released from a calcium phosphate coating to promote cell attachment and growth.

[0102] Dopants may include systemic factors that affect bone remodeling include the parathyroid hormone (PTH), calcitriol, sex hormones, glucocorticoids, and thyroid hormones, cytokines, prostaglandins, tumor growth factor-beta (TGF-β), and certain morphogenetic proteins.

[0103] Dopants and conductive substrates may include zinc, copper, magnesium, strontium, silver, gold, palladium, silicon, gallium, europium, terbium and combinations thereof.

[0104] Dopants may include bone growth proteins such as TGFβ1, TGFβ2, TGFβ3, bone morphogenic protein (BMP)-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8, BMP-9, cartilage-derived morphogenic protein (CDMP)-1, CDMP-2, and / or CDMP-3, fibroblast growth factor (FGF)-1, BMP-1, BMP-2α, BMP-2β, BMP-3b, BMP-8b, BMP-10, BMP-11, BMP-12, BMP-13, BMP-14, BMP-15, TGFβ4, TGFβ5 and combinations thereof.

[0105] Dopants and conductive substrates may include metals such as Lithium (Li), Beryllium (Be), Sodium (Na), Magnesium (Mg), Aluminum (Al), Potassium (K), Calcium (Ca), ScandiumDocket No. 2891.098AWO (Sc), Titanium (Ti), Vanadium (V), Chromium (Cr), Manganese (Mn), Iron (Fe), Cobalt (Co), Nickel (Ni), Copper (Cu), Zinc (Zn), Gallium (Ha), Rubidium (Rb), Strontium (Sr), Yttrium (Y), Zirconium (Zr), Niobium (Nb), Molybdenum (Mo), Technetium (Tc), Ruthenium (Ru), Rhodium (Rh), Palladium (Pd), Silver (Ag), Cadmium (Cd), Indium (In), Tin (Sn), Cesium (Cs), Barium (Ba), Lanthanum (La), Cerium (Ce), Praseodymium (Pr), Neodymium (Nd), Promethium (Pm), Samarium (Sm), Europium (Eu), Gadolinium (Gd), Terbium (Tb), Dysprosium (Dy), Holmium (Ho), Erbium (Er), Thulium (Tm), Ytterbium (Yb), Lutetium (Lu), Hafnium (Hf), Tantalum (Ta), Tungsten (W), Rhenium (Re), Osmium (Os), Iridium (Ir), Platinum (Pt), Gold (Au), Mercury (Hg), Thallium (Tl), Lead (PB), Bismuth (Bi), Polonium (Po), Francium (Fr), Radium (Ra), Actinium (Ac), Thorium (Th), Protactinium (Pa), Uranium (U), Neptunium (Np), Plutonium (Pu), Americium (Am), Curium (Cm), Berkelium (Bk), Californium (Cf), Einsteinium (Es), Fermium (Fm), Mendelevium (Md), Nobelium (No), Lawrencium (Lr), Rutherfordium (Rf), Dubnium (Db), Seaborgium (Sg), Bohrium (Bh), Hassium (Hs), Meitnerium (Mt), Darmstadtium (Ds), Roentgenium (Rg), Copernicium (Cn), Ununtrium (Uut), Flevorium (Fl), Livermorium (Lv) and alloys and combinations thereof. Alloys may include the combination of two or more metals.

[0106] Dopants may include antibiotics such as aminoglycoside antibiotic, an ansamycin antiobiotic, a beta-lactam antibiotic, a glycopeptide antibiotic, a lincosamide antibiotic, a lipopeptide antibiotic, a macrolide antibiotic, a monobactam antibiotic, a nitrofuran antibiotic, an oxazolidinone antibiotic, a quinolone antibiotic, a fluoroquinolone antibiotic, a sulfonamide antibiotic, a tetracycline antibiotic, pexiganan, fusidic acid, mupirocin, tobramycin, streptomycin, rifampicin, vancomycin, clindamycin, daptomycin, erythromycin, linezolid, penicillin, minocycline, pexiganan, fusidic acid, mupirocin, bacitracin, neomycin, polymixin B, metronidazole, silver, zinc, copper, and any combination thereof.

[0107] Dopants may include anti-inflammatory, pain medicines and combinations of both. Dopants may include acetylated salicylates (aspirin), non-acetylated salicylates (diflunisal, salsalate), anthranilic acids (meclofenamate and mefenamic acid), propionic acids (naproxen and ibuprofen), enolic acids (meloxicam and piroxicam), acetic acids (diclofenac and indomethacin), naphthylalanine (nabumetone), selective COX-2 inhibitors (celecoxib, etoricoxib), Oxycodone, Hydrocodone, dihydrocodeinone, Morphine, Codeine, Fentanyl, Hydromorphone, Meperidine,Docket No. 2891.098AWO Oxymorphone and combinations thereof. In some embodiments, one or more anti-inflammatory and one or more pain medicine are slowly released as the calcium phosphate coating dissolves in body fluid.

[0108] Dopants may include one or more antiproliferative, antiplatelet, anticoagulant agents such as paclitaxel, calcium channel antagonists, inhibitors of angiotensin converting enzyme, corticosteroids, biodegradable polymers, doxorubicin, vinblastine and paclitaxel.

[0109] Dopants may include one or more antirestenotic agent, antidiabetic, analgesic, antiinflammatory agent, antirheumatic, antihypotensive agent, antihypertensive agent, psychoactive drug, tranquillizer, antiemetic, muscle relaxant, glucocorticoid, agent for treating ulcerative colitis or Crohn's disease, antiallergic, antibiotic, antiepileptic, anticoagulant, antimycotic, antitussive, arteriosclerosis remedy, diuretic, protein, peptide, enzyme, enzyme inhibitor, gout remedy, hormone and inhibitor thereof, cardiac glycoside, immunotherapeutic agent and cytokine, laxative, lipid-lowering agent, migraine remedie, mineral product, otological, anti parkinson agent, thyroid therapeutic agent, spasmolytic, platelet aggregation inhibitor, vitamin, cytostatic and metastasis inhibitor, phytopharmaceutical, chemotherapeutic agent and amino acid, acarbose, antigen, beta-receptor blocker, non-steroidal antiinflammatory drug [NSAIDs], cardiac glycosides acetylsalicylic acid, virustatic, aclarubicin, acyclovir, cisplatin, actinomycin, alpha- and beta-sympatomimetics, (dmeprazole, allopurinol, alprostadil, prostaglandins, amantadine, ambroxol, amlodipine, methotrexate, S-aminosalicylic acid, amitriptyline, amoxicillin, anastrozole, atenolol, azathioprine, balsalazide, beclomethasone, betahistine, bezafibrate, bicalutamide, diazepam and diazepam derivatives, budesonide, bufexamac, buprenorphine, methadone, calcium salts, potassium salts, magnesium salts, candesartan, carbamazepine, captopril, cefalosporins, cetirizine, chenodeoxycholic acid, ursodeoxycholic acid, theophylline and theophylline derivatives, trypsins, cimetidine, clarithromycin, clavulanic acid, clindamycin, clobutinol, clonidine, cotrimoxazole, codeine, caffeine, vitamin D and derivatives of vitamin D, colestyramine, cromoglicic acid, coumarin and coumarin derivatives, cysteine, cytarabine, cyclophosphamide, ciclosporin, cyproterone, cytabarine, dapiprazole, desogestrel, desonide, dihydralazine, diltiazem, ergot alkaloids, dimenhydrinate, dimethyl sulphoxide, dimeticone, domperidone and domperidan derivatives, dopamine, doxazosin, doxorubizin, doxylamine, dapiprazole, benzodiazepines, diclofenac,Docket No. 2891.098AWO glycoside antibiotics, desipramine, econazole, ACE inhibitors, enalapril, ephedrine, epinephrine, epoetin and epoetin derivatives, morphinans, calcium antagonists, irinotecan, modafinil, orlistat, peptide antibiotics, phenyloin, riluzoles, risedronate, sildenafil, topiramate, macrolide antibiotics, oestrogen and oestrogen derivatives, progestogen and progestogen derivatives, testosterone and testosterone derivatives, androgen and androgen derivatives, ethenzamide, etofenamate, etofibrate, fenofibrate, etofylline, etoposide, famciclovir, famotidine, felodipine, fenofibrate, fentanyl, fenticonazole, gyrase inhibitors, fluconazole, fludarabine, fluarizine, fluorouracil, fluoxetine, flurbiprofen, ibuprofen, flutamide, fluvastatin, follitropin, formoterol, fosfomicin, furosemide, fusidic acid, gallopamil, ganciclovir, gemfibrozil, gentamicin, ginkgo, Saint John's wort, glibenclamide, urea derivatives as oral antidiabetics, glucagon, glucosamine and glucosamine derivatives, glutathione, glycerol and glycerol derivatives, hypothalamus hormones, goserelin, gyrase inhibitors, guanethidine, halofantrine, haloperidol, heparin and heparin derivatives, hyaluronic acid, hydralazine, hydrochlorothiazide and hydrochlorothiazide derivatives, salicylates, hydroxyzine, idarubicin, ifosfamide, imipramine, indometacin, indoramine, insulin, interferons, iodine and iodine derivatives, isoconazole, isoprenaline, glucitol and glucitol derivatives, itraconazole, ketoconazole, ketoprofen, ketotifen, lacidipine, lansoprazole, levodopa, levomethadone, thyroid hormones, lipoic acid and lipoic acid derivatives, lisinopril, lisuride, lofepramine, lomustine, loperamide, loratadine, maprotiline, mebendazole, mebeverine, meclozine, mefenamic acid, mefloquine, meloxicam, mepindolol, meprobamate, meropenem, mesalazine, mesuximide, metamizole, metformin, methotrexate, methylphenidate, methylprednisolone, metixene, metoclopramide, metoprolol, metronidazole, mianserin, miconazole, minocycline, minoxidil, misoprostol, mitomycin, mizolastine, moexipril, morphine and morphine derivatives, evening primrose, nalbuphine, naloxone, tilidine, naproxen, narcotine, natamycin, neostigmine, nicergoline, nicethamide, nifedipine, niflumic acid, nimodipine, nimorazole, nimustine, nisoldipine, adrenaline and adrenaline derivatives, norfloxacin, novamine sulfone, noscapine, nystatin, ofloxacin, olanzapine, olsalazine, omeprazole, omoconazole, ondansetron, oxaceprol, oxacillin, oxiconazole, oxymetazoline, pantoprazole, paracetamol, paroxetine, penciclovir, oral penicillins, pentazocine, pentifylline, pentoxifylline, perphenazine, pethidine, plant extracts, phenazone, pheniramine, barbituric acid derivatives, phenylbutazone, phenyloin, pimozide, pindolol, piperazine, piracetam, pirenzepine, piribedil, piroxicam, pramipexole, pravastatin, prazosin, procaine, promazine, propiverine,Docket No. 2891.098AWO propranolol, propyphenazone, prostaglandins, protionamide, proxyphylline, quetiapine, quinapril, quinaprilat, ramipril, ranitidine, reproterol, reserpine, ribavirin, rifampicin, risperidone, ritonavir, ropinirole, roxatidine, roxithromycin, ruscogenin, rutoside and rutoside derivatives, sabadilla, salbutamol, salmeterol, scopolamine, selegiline, sertaconazole, sertindole, sertralion, silicates, sildenafil, simvastatin, sitosterol, sotalol, spaglumic acid, sparfloxacin, spectinomycin, spiramycin, spirapril, spironolactone, stavudine, streptomycin, sucralfate, sufentanil, sulbactam, sulphonamides, sulfasalazine, sulpiride, sultamicillin, sultiam, sumatriptan, suxamethonium chloride, tacrine, tacrolimus, taliolol, tamoxifen, taurolidine, tazarotene, temazepam, teniposide, tenoxicam, terazosin, terbinafine, terbutaline, terfenadine, terlipressin, tertatolol, tetracyclins, teryzoline, theobromine, theophylline, butizine, thiamazole, phenothiazines, thiotepa, tiagabine, tiapride, propionic acid derivatives, ticlopidine, timolol, timidazole, tioconazole, tioguanine, tioxolone, tiropramide, tizanidine, tolazoline, tolbutamide, tolcapone, tolnaftate, tolperisone, topotecan, torasemide, antioestrogens, tramadol, tramazoline, trandolapril, tranylcypromine, trapidil, trazodone, triamcinolone and triamcinolone derivatives, triamterene, trifluperidol, trifluridine, trimethoprim, trimipramine, tripelennamine, triprolidine, trifosfamide, tromantadine, trometamol, tropalpin, troxerutine, tulobuterol, tyramine, tyrothricin, urapidil, ursodeoxycholic acid, chenodeoxycholic acid, valaciclovir, valproic acid, vancomycin, vecuronium chloride, Viagra, venlafaxine, verapamil, vidarabine, vigabatrin, viloazine, vinblastine, vincamine, vincristine, vindesine, vinorelbine, vinpocetine, viquidil, warfarin, xantinol nicotinate, xipamide, zafirlukast, zalcitabine, zidovudine, zolmitriptan, zolpidem, zoplicone, and zotipine.

[0110] The calcium phosphate coating may include at least one dopant of a particular atomic percentage. In some embodiments, the atomic percentage of the dopant within the calcium phosphate coating is about 0.05% or about 1%. In some embodiments, the atomic percentage of at least one dopant metal ion within the doped portion of the doped calcium phosphate coating is about 0.05%, about 0.1%, about 0.15%. , About 0.2%, about 0.25%, about 0.3%, about 0.35%, about 0.4%, about 0.45%, about 0.5%, about 0.55%, about 0.6%, about 0.65%, about 0.7%, about 0.75%, about 0.8%, about 0.85%, about 0.9%, about 1%, about 1.1% , About 1.2%, about 1.3%, about 1.4%, about 1.5%, about 1.6%, about 1.7%, about 1.8%, about 1.9%, about 2%, about 2.1%, about 2.2%, about 2.3%, about 2.4%, about 2.5%, about 2.6%, about 2.7%, about 2.8% , About 2.9%, about 3%, about 3.1%, about 3.2%, about 3.3%, about 3.4%, about 3.5%, about 3.6%, about 3. 7%, about 3.8%, about 3.9%, about 4%, about 4.1%, about 4.2%, aboutDocket No. 2891.098AWO 4.3%, about 4.4%, about 4.5%, about 4.6%, about 4.7%, about 4.8%, about 4.9%, about 5%, about 5.1%, about 5.2%, about 5.3%, about 5.4%, Or about 5.5%.

[0111] The calcium phosphate coating may include one or more dopant of a particular weight percentage of the calcium phosphate coating. In some embodiments, the weight percentage of the dopant within the calcium phosphate coating is about 0.1 percent weight , about 0.2 percent weight , about 0.3 percent weight , about 0.4 percent weight , about 0.5 percent weight , about 0.6 percent weight , about 0.7 percent weight , about 0.8 percent weight , about 0.9 percent weight , about 1 percent weight , about 2 percent weight , about 3 percent weight , about 4 percent weight , about 5 percent weight , about 6 percent weight , about 7 percent weight , about 8 percent weight , about 9 percent weight , about 10 percent, about 11 percent, about 12 percent, about 13 percent, about 14 percent, about 15 percent, about 16 percent, about 17 percent, about 18 percent, about 19 percent, about 20 percent, about 21 percent, about 22 percent, about 23 percent, about 24 percent, about 25 percent, about 30 percent, or about 50 percent weight of the calcium phosphate coating.

[0112] As used herein, the term “about” means that the numerical value is approximate and small variations would not significantly affect the practice of the disclosed embodiments. Where a numerical limitation is used, unless indicated otherwise by the context, “about” means the numerical value can vary by ±1 or ±10% , or any point therein, and remain within the scope of the disclosed embodiments.

[0113] As used herein, embodiments and embodiments may be used interchangeably to describe non-limiting embodiments. EXAMPLES

[0114] The following examples are intended to illustrate particular embodiments of the present disclosure, but are not intended to limit the scope thereof.

[0115] Although some non-limiting examples have been depicted and described in detail herein, it will be apparent to those skilled in the relevant art that various modifications, additions, substitutions, and the like can be made without departing from the spirit of the present disclosureDocket No. 2891.098AWO and these are therefore considered to be within the scope of the present disclosure as defined in the claims that follow.

[0116] It should be appreciated that all combinations of the foregoing concepts and additional concepts discussed in greater detail herein (provided such concepts are not mutually inconsistent) are contemplated as being part of the inventive subject matter disclosed herein. In particular, all combinations of claimed subject matter appearing at the end of this disclosure are contemplated as being part of the inventive subject matter disclosed herein and may be used to achieve the benefits and advantages described herein.

[0117] Conductive substrates that may be used for nanometer thick calcium phosphate coatings may utilize one or more surface preparation prior to the nano second electrodeposition of the coating. a. In some embodiments, one or more surface preparation may include: Passivation, Plasma cleaning, Ultrasonic Cleaning, Acid Etching, Anodization and dip coating. In some embodiments, additional coatings may be applied to the conductive substrate, the calcium phosphate coating or any combination thereof. Sodium silicate may be an additional coating applied to the conductive substrate using electric nanosecond pulses. Sodium silicate may be added to a one or more calcium phosphate coatings, the conductive substrate or both. Sodium silicate may be added to one or more layers of calcium phosphate coatings, the conductive substrate or both by dipping, nanosecond electrodeposition or a combinations thereof. In some embodiments, sodium silicate may be a dopant.

[0118] In some embodiments, there is preparation of deposition solution for pulsed nanosecond electrodeposition for example: a. Solution containing Ca+ and PO4- Ions along with any dopants. Configurations currently use either CaCl2 or CaNO3 with NH4H2PO4. For a zinc dopant we used ZnCl2 and Zn(NO3)2, Copper nitrate or chloride

[0119] In some embodiments, a calcium phosphate coating on a conductive substrate may include post-processing such which may include:Docket No. 2891.098AWO a. Chemical base or alkali may be in a solution, such as sodium or ammonium hydroxide, soak at various temperatures and solution concentrations. Warmer base or alkali solution, warmer environment may be used to increase conversion speed and make smaller calcium phosphate crystals. Colder base or alkali solution may be used for larger calcium phosphate crystals. Higher concentration of alkali or base may be used for smaller calcium phosphate crystals and lower concentration base or alkali may be used for larger calcium phosphate crystals. b. Thermal or hydrothermal treatment may be used to achieve various forms or sizes of calcium phosphate crystals. Thermal annealing may be used. In some embodiments, thermal annealing may be at about 400 degrees centigrade, 500 degrees centigrade, 800 degrees centigrade or 1000 degrees centigrade. c. Chemical bath or soak may be used to incorporate other ions, antimicrobials, drugs, biologics, stains, dyes, DNA, RNA or vector (liposome, virus, etc.) In some embodiments, the chemical bath or soak may be used to diffuse dopants into the calcium phosphate coating. d. Additional amorphous CaP coating may be applied to protect the surface from contamination. e. Chemical or Plasma vapor deposition may be used to such as for the addition of dopants. f. LASER treatment may be used to change, ablate or convert complete or localized areas of the surface. g. Plasma treatment may be used to activate the surface for additional treatments or coatings.

[0120] In some embodiments, a nanometer thick crystalline calcium phosphate coating includes the following procedure: 1. Clean Conductive Substrate. a. Ultrasonic clean dilute lab soap solution 1hr. b. Rinse well and dry. 2. Create Calcium Phosphate Deposition SolutionDocket No. 2891.098AWO a. Create 100x CaCl2Stock.b.Create 100x NH4H2PO4Stock. c. In 500ml Beaker add 498ml of H2O with stir bar. d. Add NaCl to 500ml of H20 while vigorously stirring. e. Add CaCl2 stock. f. Add NH4H2PO4 stock slowly dropwise to limit precipitation. g. Turn off stirring. 3. Setup Electrode Array (Fig 1) a. Secure Monel Anode with Ring Stand (MAKE SURE RUBBER TIPS ON CALMPS AND ANODE IS ELECTRICLY INSULATED) b. Attach sample holder (Fig 2) c. Rest on Electrical Contact 4. Pulse a. Make sure solution is stagnant. b. Apply desired pulse amplitude (example pulse Fig 3). c. During pulsing make sure system is properly isolated and proper. electrical contact is made, look for sparking. d. Check for color consistency on substrate for desired thickness of coating based approximately on color. e. Remove sample, rinse in H2O, and blow dry. 5. Amorphous calcium phosphate to Crystalline calcium phosphate Conversion a. Submerge Samples in NaOH at desired concentration and temperature. b. Remove sample, rinse in H2O, and blow dry (color should be gone.)Docket No. 2891.098AWO 6. Imaging: use 5KV, 10mA to get clear imagine and minimize melting of HA Parameters, where P-count is the total number of nanosecond electric pulses, where amplitude is the voltage, NaOH is the base used to convert amorphous nanometer thick calcium phosphate coating to nanometer thick crystalline calcium phosphate coating where the crystals have a vertical orientation. Sample CaCl2NH4H2PO4NaCl P-Count Amplitude NaOH Conversion

[0120] FIG. 1A shows a non-limiting embodiment of cathodic electrodeposition of nanometer thick amorphous calcium phosphate (CaP) on a conductive substrate in an electrolyte at room temperature, where the substrate is electrically connected to the negative electrode and is negatively charged during the nanosecond electric pulse of direct current. In this embodiment the electrolyte contains calcium and phosphate as salts, CaCl2 and NH4H2PO4, respectively. The dissolved salts dissociate into charged ions wherein the charged calcium and phosphate ions are ionically bonded to the charged conductive substrate. In this embodiment a non-conductive container holds the electrolyte, conductive substrate and electrodes such that they are connected electrically when the electric pulse is delivered to the electrode. This is an embodiment of cathodic electrodeposition of a nanometer thick calcium phosphate coating on a conductive substrate. In some embodiments, the conductive substrate is connected to the positive electrode. In some embodiments, one or more conductive substrates are connected to one or more negative electrode. In some embodiments, one or more conductive substrates are connected to one or more positive electrode. In some embodiments, one or more dopant is co-electrodeposited with calcium phosphate. In some embodiments, the conductive substrate has a pre-existing nanometer size topology. In some embodiments, the electrolyte contains calcium phosphate plus at least oneDocket No. 2891.098AWO additional dopant. In some embodiments, the nanometer thick amorphous calcium phosphate is electrodeposited on a conductive substrate and then converted by a base to crystalline calcium phosphate. In some embodiments, a nanometer size topology is applied to a conductive substrate by the addition of nanometer thick crystalline calcium phosphate coating.

[0121] FIG. 1B shows a non-limiting embodiment of a conductive substrate, a stainless steel washer, electrically connected to an approximately three inch supporting wire that holds the substrate in the electrolyte during calcium phosphate coating. In some embodiments, the substrate acts as the electrode. In some embodiments, the substrate has additional non-conductive regions that do not get coated. In some embodiments, one or more substrates are electrically connected together. In some embodiments, a counter electrode contains dopants that are co- deposited with the nanometer thick calcium phosphate coating. In some embodiments, the conductive substrate is masked in one or more region to prevent coating. In some embodiments the conductive substrate has a negative charge. In some embodiments the conductive substrate has a positive charge. In some embodiments one or more substrate has a first negative charge and a second positive charge, a first positive charge and a second negative charge or any combination thereof.

[0122] FIG. 2 shows a non-limiting embodiment of the measured output of a nanosecond electric pulse generator configured to deliver a 4kv nanosecond electric pulse. A pulse generator may be used to deliver nanosecond pulses of direct electrical current to a conductive substrate for the electrodeposition of nanometer thick calcium phosphate coatings. A nanometer thick amorphous calcium phosphate coating may be deposited on a conductive substrate using a nanosecond pulse generator. In some embodiments, the nanosecond electric pulse generator may deliver a nanosecond electric pulse of about 1 to 40 kilovolts. In some embodiments, the pulse generator may deliver nanosecond electric pulses at about 0.001 hertz. In some embodiments, the pulse generator may deliver nanosecond electric pulses at about 0.01 hertz. In some embodiments, the pulse generator may deliver nanosecond electric pulses at about 0.1 hertz. In some embodiments, the pulse generator may deliver nanosecond electric pulses at about 1 hertz. In some embodiments, the pulse generator may deliver nanosecond electric pulses at about 10 hertz. In some embodiments, the pulse generator may deliver nanosecond electric pulses at about 100 hertz. In some embodiments, the pulse generator may deliver nanosecond electric pulses at aboutDocket No. 2891.098AWO 1000 hertz. In some embodiments, the pulse generator may deliver electric pulses at about 100,000 hertz. In some embodiments, the number of total number of nanosecond electric pulses may be varied. In some embodiments, one or more voltage, hertz and total electric pulses may be varied to deliver a predetermined nanometer thick calcium phosphate coating.

[0123] FIG. 3 shows a non-limiting embodiment of a conductive substrate, in this embodiment a 316L stainless steel washer. The washer is coated with nanometer thick amorphous calcium phosphate as the intermediate step to creating the 75 nanometer small texture crystalline surface (FIG. 4). The amorphous layer is approximately 150 nanometers thick though not shown in the image. The colorful appearance on the washer is from the uniform nanometer thick film which causes light interference. In some embodiments, a small texture may be used to describe a crystalline calcium phosphate surface with small crystals of about 1 to about 250 nanometers. In some embodiments, a large texture may be used to describe a crystalline calcium phosphate surface with large crystals of about 300 to about 1000 nanometers.

[0124] FIG. 4 shows a non-limiting embodiment of an SEM image of nanometer thick amorphous calcium phosphate electrodeposition on stainless-steel washer from FIG. 3 then post processed to a crystalline state. The electrodeposition liquid was aqueous CaCl25mM, NH4H2PO4 3mM and NaCl 50mM. The electric pulse voltage was 4,000 volts. The electric pulse duration was 300 nanoseconds. The total number of pulses was 1500 applied at 5-10hz. The washer was electrically connected to the cathode and thereby negatively charged. After electrodeposition the substrate was treated in a bath of NaOH 0.5M for 2 hours at 65°C. This sodium hydroxide treatment converts amorphous calcium phosphate coating to crystalline calcium phosphate coating. In some embodiments, the quicker the nanometer thick amorphous calcium phosphate coating is converted using a base the smaller nanometer calcium phosphate crystals become. In some embodiments, using a stronger heated base converts the nanometer thick amorphous calcium phosphate quicker and in turn into smaller nanometer sized calcium phosphate crystals. In some embodiments, modifying the temperature of the base and strength of base can determine how quickly nanometer thick amorphous calcium phosphate is converted to nanometer thick crystalline calcium phosphate and the nanometer size of the calcium phosphate crystals. In some embodiments, the size of the calcium phosphate crystals is dependent on the alkaline soak time, alkaline temperature, alkaline strength, pH and combinations thereof. In someDocket No. 2891.098AWO embodiments, the crystal size depends on the rate of crystal formation. In some embodiments, higher temperatures of a basic pH solution convert the amorphous calcium phosphate to crystalline calcium phosphate faster or at a higher rate and lead to smaller crystals. In some embodiments, a higher concentration of basic pH solution converts the amorphous calcium phosphate to crystalline calcium phosphate faster or a higher rate leading to smaller crystals. In some embodiments, the higher the concentration of base in a solution the higher the pH and the more quickly amorphous calcium phosphate is converted to crystalline calcium phosphate. In some embodiments, both higher concentrations and heated basic pH solution together convert the amorphous calcium phosphate to crystalline calcium phosphate more quickly and produce smaller crystals. In embodiments, the amount of time used in the conversion of amorphous calcium phosphate to crystalline calcium phosphate determines the final crystal size. A longer conversion time may result in larger calcium phosphate crystals forming from the amorphous calcium phosphate layer. A shorter conversion time may result in smaller calcium phosphate crystals being formed from the amorphous calcium phosphate layer. In some embodiments, a partially converted amorphous calcium phosphate layer on a surface results in a semicrystalline calcium phosphate layer on a surface.

[0125] FIG. 5 shows a non-limiting embodiment of a nanometer thick amorphous calcium phosphate coating on a stainless steel washer.

[0126] FIG. 6 shows a non-limiting embodiment of an SEM image of nanometer thick crystalline calcium phosphate on stainless-steel. The crystals are approximately 100 nanometers wide by 200 nanometers tall. The calcium phosphate was electrodeposited and the electrodeposition liquid was aqueous CaCl25mM, NH4H2PO43mM and NaCl 50mM. The electric pulse voltage was 4,000 volts. The electric pulse duration was 300 nanoseconds. The total number of nanosecond electric pulses was 2500. The substrate was electrically connected to the cathode and thereby negatively charged. After electrodeposition the substrate was treated in a bath of NaOH 0.05M for 2 hours at 65°C. This sodium hydroxide treatments converts the nanometer thick amorphous morphology calcium phosphate to a crystalline nanoscale morphology. The nanoscale morphology of the electrodeposition material is larger than FIG. 4 which has similar conditions except for the total count of electric pulses was 1500 in FIG. 4 and the concentration of the post processing NaOH was ten-fold higher in FIG. 4. The largerDocket No. 2891.098AWO morphology includes larger crystal size and topology. The larger topology of the surface having altered biological properties compared to FIG. 4.

[0127] FIG. 7 shows a non-limiting embodiment of an SEM image of a stainless steel substrate that is used as control and has no surface coating. The stainless steel surface contains approximately 500 nanometer wide grain boundaries that represent the pre-existing nanometer topology of the substrate. In some embodiments, nanometer scale topologies include nanotubes. In some embodiments, nanometer scale topologies include nanoparticles. In some embodiments, nanometer scale topologies include nanopores. In some embodiments, one or more nanometer size topology is included on a substrate. Nanoscale topology includes features, objects, and particles that have at least one dimension that is nanometer in size. In some embodiments, a substrate may have a pre-existing topology that is micron sized wide but nanometer in height and may be considered to have a nanoscale topology. In some embodiments, nanoscale refers to all dimensions being nanometer in size.

[0128] FIG. 8 shows a non-limiting embodiment of an SEM image of a cathodic deposition of nanometer thick calcium phosphate onto a stainless-steel substrate. Stainless-steel was electrically connected to the cathode and thereby negatively charged during the nanosecond electrical pulses. The electric pulse voltage was 4000 volts, the electric pulse duration was 300 nanoseconds, the electric pulse period was 5 hertz, and the total number of electric pulses was 2000. The concentration of the electrodeposition liquid was aqueous 25 mM CaCl2, 3 mM NH4H2PO4, and 50 mM NaCl. The NaCl acts as a bystander ion to adjust the solution conductivity. The nanometer thick amorphous calcium phosphate layer has an approximate 200 nm thickness with a texture less than 200 nm. Texture may include distances from the average coating height to the average coating depth. The 500 nanometer wide grain boundaries can still be observed on the stainless steel surface and the nanometer thick amorphous coating preserves the underlying surface topology. In some embodiments, dopants are included in the nanometer thick amorphous calcium phosphate coating. The grain boundaries are coated in amorphous calcium phosphate and the nanometer topology of the stainless steel is preserved. In some embodiments, stainless steel is not biologically functional such that it does not promote cell adhesion, proliferation or differentiation and stainless steel modified with a calcium phosphateDocket No. 2891.098AWO coating will promote osteoblast adhesion and proliferation. In some embodiments, dopants included in the calcium phosphate coating prevent infections and promote cell growth.

[0129] FIG. 9A-9B shows non-limiting embodiments of SEM images demonstrating the conversion of nanometer thick amorphous calcium phosphate (ACP) to nanometer thick crystalline calcium phosphate by soaking the ACP in a basic pH solution for a period of time. The left panel, FIG. 9A, shows small nanocrystals on the ACP surface. The right panel , FIG. 9B, shows large nanocrystals after the ACP surface has been soaked in the basic pH solution converting the small ACP nanocrystals to larger nanocrystals. The small calcium phosphate crystal size is made with a thinner ACP layer with a higher concentration heated base soak. The large calcium phosphate crystals are made with a thicker ACP layer then converted slower with a lower concentration cooler base soak. The thinner ACP layer allows for the crystal density and orientation to remain the same when varying size. In some embodiments, the crystal density and orientation may be determined by the thickness of the ACP coating.

[0130] FIG. 10 shows a non-limiting embodiment of an SEM image of a substrate with TiO2 nanotubes without a nanometer thick calcium phosphate coating. The TiO2 nanotubes are approximately 30-90nanometers in width and are an example of a preexisting nanometer scale topology on a substrate.

[0131] FIG. 11 shows a non-limiting embodiment of an SEM image of nanometer thick amorphous zinc calcium phosphate (Zn-CaP) on the nanoscale TiO2 nanotubes of FIG. 10. The preexisting nanometer scale topology of the TiO2 nanotubes was preserved and the nanotubes can still be seen. The nanometer thick amorphous calcium phosphate was doped with zinc which was co-deposited with the calcium phosphate. Using the aqueous solution of zinc adds new properties to the nanometer thick calcium phosphate coating such as enhancing osteoblast function and acting as an antimicrobial. In some embodiments, the concentration of zinc may determine the strength of the antimicrobial properties of the surface.

[0132] FIG. 12 shows a non-limiting embodiment of an SEM image of nanometer thick amorphous zinc calcium phosphate (Zn-CaP) converted to nanometer thick crystalline zinc calcium phosphate (Zn-CaP) on 30-90 nm titanium anodized nanotubes from FIG. 11. The preexisting nanometer scale topology of the TiO2 nanotubes is maintained even after the conversion of nanometer thick amorphous zinc calcium phosphate to nanometer thick crystallineDocket No. 2891.098AWO zinc calcium phosphate. The crystalline calcium phosphate adds additional nanoscale texture, while being more resistant to dissolution as compared to an amorphous layer.

[0133] FIG. 13 shows a non-limiting embodiment of Alizarin red quantification on titanium surfaces 21 days post differentiation of mesenchymal stem cells (MSCs). Alizarin red is a stain used to identify calcium containing osteocytes in differentiated MSCs. The absorbance at 490 nanometer wavelength corresponds to the amount of signal from Alizarin red which directly correlates to the amount of calcium rich boney mineral matrix on a specific surface due to MSC differentiating into matrix producing osteocytes. The FIG. shows the untreated control surface has the least osteocytes on titanium while the addition of a nanometer thick coatings of amorphous calcium phosphate with nanometer scale topology on the titanium result in greater osteocyte differentiation. An embodiment of the nanotubes morphology with a nanometer thick amorphous calcium phosphate coating as used in this embodiment can be found in FIG. 11. The addition of nanometer thick zinc calcium phosphate to the nanotubes shows a further increase in the number of osteocytes on the surface with osteocytes increasing with concentration of zinc in the surface coating. In some embodiments, other dopants may be used.

[0134] FIG. 14 shows a non-limiting embodiment of Alizarin red quantification on stainless steel surfaces 21 days post differentiation of mesenchymal stem cells (MSCs). The absorbance at 490 nm wavelength corresponds to the amount of signal from Alizarin red which directly correlates to the amount of calcium rich boney mineral matrix on a specific surface due to MSC differentiating into matrix producing osteocytes. The control stainless steel surface without a nanometer thick crystalline calcium phosphate coating has the lowest number of osteocytes as indicated by absorbance at 490 nm while the nanometer thick crystalline octacalcium phosphate has the highest Alizarin red signal followed by nanometer thick crystalline octacalcium phosphate with 10% zinc and crystalline octacalcium phosphate with 1% zinc. The results demonstrate nanometer thick crystalline octacalcium phosphate coating is a preferable environment for osteocytes but the inclusion of 1% to 10% zinc is still preferable for osteocyte growth than a surface without a nanometer thick crystalline calcium phosphate coating.

[0135] FIG. 15 shows a non-limiting embodiment of a conductive substrate 200 with a plurality of calcium phosphate nanocrystals 220 where the vertical orientation of the crystals 220 is relative to the conductive substrate 200 with the greatest vertical orientation being a 90 degreeDocket No. 2891.098AWO angle perpendicular 230 to the substrate 200 as measured by a central axis of the crystal. In some embodiments, crystals have a 90 degree vertical orientation 240 to the substrate 200. In some embodiments, a plurality of calcium phosphate crystals, or a proportion or percentage thereof, have a minimum vertical angle 210 as measured from the substrate 200 to a central axis 205 of the crystals 220. In some embodiments, a plurality of calcium phosphate crystals have a minimum vertical angle 210 of at least 22. 5 degrees. In some embodiments, a plurality of calcium phosphate crystals have a minimum vertical angle of at least 20 degrees. In some embodiments, about 50 percent of the plurality of the crystals have a minimum vertical angle of at least 22.5 degrees. In some embodiments, about 75 percent of the plurality of crystals have a minimum vertical angle of at least 22.5 degrees. In some embodiments, about 95 percent of the plurality of crystals has a minimum vertical angle of at least 22.5 degrees.

[0136] FIG. 16 shows a non-limiting embodiment of dopants in the form of ions that may be included in calcium phosphate coatings as described herein. The figure provides embodiments of examples of concentrations of ions demonstrated to have a biological effect as described. Higher or lower concentrations may be used. Combinations of ions and other dopants may be used. Gradients or layers of dopants may be used.

[0137] FIG. 17 shows a non-limiting embodiment of low density crystals of calcium phosphate. Crystal density as described herein refers to the numbers of crystals per unit of area. As seen by comparing FIGs 17 and 18 the density of crystals can be tuned or controlled based on the nanometer thickness of the amorphous calcium phosphate coating and the conversion process used. FIG. 17 has a thinner nanometer thick amorphous calcium phosphate layer as compared to FIG. 18 and has fewer crystals per square micrometer. In some embodiments electrodepositing about 100-250 nanometers of amorphous calcium phosphate results in less crystal density than electrodepositing about 250-500 nanometers of amorphous calcium phosphate, In some embodiments electrodepositing 1-50 nanometers of amorphous calcium phosphate leads to less dense vertical crystals than electrodepositing 50-100 nanometers of amorphous calcium phosphate.

[0138] FIG. 18 shows a non-limiting embodiment of high density crystals of calcium phosphate. The thickness of the amorphous calcium phosphate layer may determine the density of calcium phosphate crystals on a surface, where the density of crystals as described herein refers to theDocket No. 2891.098AWO numbers of crystals per unit of area. In some embodiments, a thicker nanometer amorphous calcium phosphate layer on a surface will be converted into more densely clustered nanometer calcium phosphate crystals. In other embodiments, a thinner nanometer layer of amorphous calcium phosphate will be converted to less dense clusters of nanometer thick calcium phosphate crystals. Electrodeposition of a thicker nanometer amorphous calcium phosphate coating allows for the conversion process to produce more densely packed nanometer size calcium phosphate crystals. In some embodiments, the thicker the nanometer thick amorphous calcium phosphate coating the larger more dense crystals can be created during the conversion process. In some embodiments, the relatively thicker amorphous calcium phosphate coating is capable of larger more dense calcium phosphate crystals, but a quick conversion process can produce a less dense smaller calcium phosphate crystal. In some embodiments electrodepositing 100-250 nanometers of amorphous calcium phosphate leads to more dense nanometer calcium phosphate crystals the electrodepositing 1-50 nanometers of amorphous calcium phosphate. In some embodiments high density calcium phosphate nanocrystals of FIG. 18 have a greater vertical orientation that less dense calcium phosphate nanocrystals FIG.17. In some embodiments increasing the density of calcium phosphate nanocrystals increases the vertical orientation about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22, about 23, about 24, about 25, about 26, about 27, about 28, about 29 or about 30 degrees from the surface of the substrate as compared to a less dense calcium phosphate nanocrystal surface. In some embodiments increasing the crystal density increases the minimum vertical orientation angle.

[0139] FIG. 19 shows a non-limiting embodiment of crystalline calcium phosphate on a stainless steel surface. In this embodiment amorphous calcium phosphate was converted to crystalline calcium phosphate using a basic solution on the stainless steel surface. The grain boundaries can still be seen though they are coated with crystalline calcium phosphate. The nanometer topology of the grain boundaries is still present but much less pronounced due to the calcium phosphate coating. A plurality of vertical nanometer crystals can be seen on the surface.

[0140] The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the disclosure. As used herein, the singular forms “a”, “an” and “the” are intended to include the plural forms as well, unless the context clearlyDocket No. 2891.098AWO indicates otherwise. It will be further understood that the terms “comprise” (and any form of comprise, such as “comprises” and “comprising”), “have” (and any form of have, such as “has”, and “having”), “include” (and any form of include, such as “includes” and “including”), and “contain” (and any form of contain, such as “contains” and “containing”) are open-ended linking verbs. As a result, a method or device that “comprises,” “has,” “includes,” or “contains” one or more steps or elements possesses those one or more steps or elements, but is not limited to possessing only those one or more steps or elements. Likewise, a step of a method or an element of a device that “comprises,” “has,” “includes,” or “contains” one or more features possesses those one or more features, but is not limited to possessing only those one or more features. Furthermore, a device or structure that is configured in a certain way is configured in at least that way, but may also be configured in ways that are not listed.

[0141] The disclosure has been described with reference to the preferred embodiments. It will be understood that the architectural and operational embodiments described herein are exemplary of a plurality of possible arrangements to provide the same general features, characteristics, and general system operation. Modifications and alterations will occur to others upon a reading and understanding of the preceding detailed description. It is intended that the disclosure be construed as including all such modifications and alterations.

Claims

Docket No. 2891.098AWO CLAIMS What is claimed:

1. A calcium phosphate coating comprising a plurality of nanometer sized calcium phosphate crystals, wherein the crystals form a single layer, wherein the coating is on a surface of a conductive substrate and is about 1 to 1000 nanometers thick, and wherein greater than 25 percent of the coated surface area comprises the single layer of the crystals.

2. The calcium phosphate coating of claim 1, where the conductive substrate is metal.

3. The calcium phosphate coating of claim 1, wherein greater than 50 percent of an exterior surface comprises the single layer of the crystals.

4. The calcium phosphate coating of claim 1, wherein greater than 50 percent of the crystals are vertically orientated about 20 degrees or greater from the surface.

5. The calcium phosphate coating of claim 1, wherein the crystals are ionically bound to the substrate.

6. The calcium phosphate coating of claim 1, wherein the crystals are created from a layer of amorphous calcium phosphate 1 to 1000 nanometers thick on the substrate.

7. The calcium phosphate coating of claim 1, wherein about 75 percent of the crystals are vertically orientated about 20 degrees or greater from the surface.

8. The calcium phosphate coating of claim 1, wherein about 95 percent of the crystals are vertically orientated about 20 degrees or greater from the surface.

9. The calcium phosphate coating of claim 1, wherein a density of the crystals is determined by a thickness of the coating.Docket No. 2891.098AWO 10. The calcium phosphate coating of claim 6, wherein a size of the crystals is determined by a rate of conversion of amorphous calcium phosphate to crystalline calcium phosphate and a total conversion time.

11. The calcium phosphate coating of claim 1, wherein the crystals are about 1 to 500 nanometers in length.

12. The calcium phosphate coating of claim 1, wherein the crystals have an aspect ratio of about 1:1.5 to 1:

10.

13. The calcium phosphate coating of claim 1, wherein the coating is about 1 to about 100 nanometers thick.

14. The calcium phosphate coating of claim 1, wherein the coating is about 150 to about 1000 nanometers thick.

15. The calcium phosphate coating of claim 1, wherein the coating is about 100 to about 500 nanometers thick.

16. The calcium phosphate coating of claim 1, wherein the coating is about 500 to about 1000 nanometers thick.

17. The calcium phosphate coating of claim 1, wherein the calcium phosphate comprises hydroxyapatite, calcium deficient hydroxyapatite, octacalcium phosphate, dicalcium phosphate dihydrate, tricalcium phosphate and brushite.

18. The calcium phosphate coating of claim 1, wherein the coating is adhered to the substrate at less than 100 degrees centigrade.

19. The calcium phosphate coating of claim 1, wherein the calcium phosphate coating is adhered to the substrate by electrodeposition in an aqueous solution.Docket No. 2891.098AWO 20. The calcium phosphate coating of claim 1, wherein the calcium phosphate coating is adhered to the substrate by nanosecond electric pulse electrodeposition in an aqueous solution.

21. The calcium phosphate coating of claim 1, wherein the substrate comprises an engineered nanometer scale topology and wherein the calcium phosphate coating preserves the nanometer scale topology of the substrate.

22. The calcium phosphate coating of claim 1, wherein the substrate comprises an engineered micron scale topology and wherein the calcium phosphate coating preserves the micron scale topology of the substrate.

23. The calcium phosphate coating of claim 1, wherein one or more dopants are adhered to the substrate with the calcium phosphate coating.

24. The calcium phosphate coating of claim 1, wherein one or more dopants are electrodeposited with the calcium phosphate coating to substrate.

25. The calcium phosphate coating of claim 1, wherein one or more dopants are diffused into the calcium phosphate coating.

26. The calcium phosphate coating of claim 1, wherein nanoparticles are added interstitial to a calcium phosphate layer or layers.

27. The calcium phosphate coating of claim 1, wherein nanoparticles are co-deposited with the calcium phosphate coating onto the substrate.

28. The calcium phosphate coating of claim 1, wherein the conductive substrate comprises nanoparticles prior to addition of the calcium phosphate coating.

29. The calcium phosphate coating of claim 1, wherein nanoparticles are added to the coating after the calcium phosphate coating.Docket No. 2891.098AWO 30. The calcium phosphate coating of claim 23, wherein the one or more dopants comprise copper, magnesium, manganese, strontium, zinc, potassium, silver, gold, sodium, fluoride, chlorine, bromine, ions of the foregoing, or any combination thereof.

31. The calcium phosphate coating of claim 1, wherein the calcium phosphate coating dissolves at a predetermined time, rate or combinations of both.

32. The calcium phosphate coating claim 1, wherein the calcium phosphate coating dissolves at a predetermined time to release one or more dopant, nanoparticle or combinations of both.

33. The calcium phosphate coating of claim 1, wherein the calcium phosphate coating dissolves at a predetermined rate to release one or more dopant, nanoparticle or combinations of both.

34. The calcium phosphate coating of claim 23, wherein the one or more dopants comprise DNA, RNA, vitamins, minerals, antimicrobials, antivirals, steroids, anti-inflammatories, pain relief medication and growth factors.

35. The calcium phosphate coating of claim 27, wherein the nanoparticles comprise DNA, RNA, vitamins, minerals, antimicrobials, antivirals, steroids, anti-inflammatories, pain relief medication and growth factors.

36. The calcium phosphate coating of claim 1, wherein dissolution of the calcium phosphate coating releases antimicrobials.

37. The calcium phosphate coating of claim 1, wherein at least one additional amorphous calcium phosphate layer is deposited on the coating.

38. The calcium phosphate coating of claim 1, wherein at least one additional amorphous calcium phosphate layer comprising dopants is deposited on the coating.Docket No. 2891.098AWO 39. The calcium phosphate coating of claim 1, wherein at least one additional amorphous calcium phosphate layer comprising biological agents is deposited on the coating.

40. The calcium phosphate coating of claim 37, wherein the at least one additional amorphous calcium phosphate layer dissolves after a predetermined period of time and the calcium phosphate coating dissolves at a second predetermined period of time after the amorphous calcium phosphate layer.

41. The calcium phosphate coating of claim 24, wherein the one or more dopant deposited with the calcium phosphate determines a dissolution rate of one or more coating.

42. The calcium phosphate coating of claim 1, wherein the coating promotes mesenchymal stem cell differentiation into osteoblasts.

43. The calcium phosphate coating of claim 1, wherein the coating includes at least one dopant which further promotes mesenchymal stem cell differentiation into osteoblasts as compared to the calcium phosphate coating alone.

44. The calcium phosphate coating of claim 1, wherein the coating contains one or more fluorescent dye, photoluminescent dye, radiochemical or combinations thereof.

45. The calcium phosphate coating of claim 1, wherein the coating comprises a first nanometer thick layer of amorphous calcium phosphate coated on the conductive substrate, wherein the amorphous calcium phosphate coating is converted to crystalline calcium phosphate using a solution with a basic pH.

46. The calcium phosphate coating of claim 1, wherein the coating comprises a first nanometer thick layer of amorphous calcium phosphate coated on the conductive substrate, wherein the amorphous calcium phosphate coating is converted to crystalline calcium phosphate using a solution with a basic pH, wherein the crystalline calcium phosphate comprises nanometer sized calcium phosphate crystals and wherein the size of the plurality of nanometer calciumDocket No. 2891.098AWO phosphate crystals is determined a pH of the solution, a temperature of the solution and a time the first nanometer thick layer of amorphous calcium phosphate soaks in the solution.

47. An amorphous calcium phosphate coating, wherein the coating is on a surface of a conductive substrate and is about 1 to 1000 nanometers thick, and wherein the coating is characterized by a nanometer surface roughness wherein the nanometer surface roughness can come from a roughness of the conductive substrate, a roughness of the amorphous calcium phosphate coating or a combination of both.

48. The amorphous calcium phosphate coating of claim 47, wherein the amorphous calcium phosphate is deposited on the conductive substrate using nanosecond electric pulses.

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