Method of biomedical material production and material produced thereunder

FIELD: medicine.

SUBSTANCE: method of biomedical material production based on titanium and cobalt alloy in mode CBC includes as follows. Exothermal mixture of raw reagents made of powdered titanium and cobalt is prepared, added with max. 4 wt % titanium hydride and max. 15 wt % amorphous nanodisperse powdered calcium hydroxyapatite (CHA) or nanocomposite calcium hydroxyapatite with biopolymers. This powdered raw mixture is pressed to preparation blanks placed to reactor CBC. Preparation blank is preheated to temperature 350-580°C. Combustion action is initiated in inert atmosphere with following release of end product. Produced biomedical material represents mushy alloy of formula TiCo of total porosity 55-70% at open porosity portion 90-98% with prolate pores sized 200-800 mcm. Pore partitions are also porous with prevailing round pores of diameter ca 100 microns. Surface of mushy alloy is coated with calcium, phosphor and oxygen compounds which are hydroxyapatite decomposition products with prevailing content of calcium perovskite.

EFFECT: material is noncytotoxic, cells are characteristically split and actively migrate inside of mushy space.

4 cl, 4 dwg, 3 ex

 

The invention relates to medicine, in particular to methods for new porous biomedical materials based on alloy titanium-cobalt, which can be used for the manufacture of bone implants.

Design and development of new materials for bone implants remains a challenge for modern implantology and restorative dentistry. Further progress in this area is associated with the development of high technologies of new materials for implants. Recently in medicine, more and more applications get composite materials that combine the properties of metals, polymers and ceramics. At the same time with this particular interest are porous materials, which by their structure and properties would be best suited bone. On the one hand, porous materials ensure the process of osseointegration, with another possible use of such materials as carriers of cellular material, primarily stem cells (D.I.Ilan, A.L.Ladd: Bone Graft Substitutes, Operative Techniques in Plastic and Reconstructive Surgery, 2003, vol.9, pp.151-160; S.J.Simske, R.A.Ayers, T.A.Bateman: Porous materials for bone engineering, Mater Sci Forum, 1997, vol.250, pp.151; T.W.Bauer, G.F.Muschler: Bone graft materials: an overview of the basic science, Clin Orthop Rel Res, 2000, vol.371, pp.10-27). One such promising materials for implants are alloys based on titanium-cobalt is.

Alloys based on cobalt are widely used in prosthetic dentistry for the manufacture of castings, frames dentures (EN 2009247 C1, C22C 19/07, 15.03.1994), for the manufacture of metal parts dentures (EN 2021882 C1, B22F 3/12, SS 1/04, SS 19/07, AK 6/04, 30.10.1994; EN 2057492 C1, A61C 13/083, 10.04.1996). The alloys are of great interest for medicine because they have a high strength, corrosion resistance and low wear (D.Granch, et al., Biomater., 1999, 20, 1079). Despite these positive characteristics cobalt alloys, in comparison with titanium, have a lower biological compatibility. Upon implantation in the body of the implant made of cobalt alloys, formed around them is not a viable granular tissue, blood tissue data is difficult, which leads to rapid release of the implant (Igolkin A.I. Titanium in medicine. Titanium, 1993, No. 1, pp.86-90, M VILS). In order to avoid negative effects, in particular, cobalt in a living organism there are different ways. One of such methods is the application by plasma deposition of bioactive coatings on the surface of a metal (cobalt containing) materials. (Yanke S.J., Pletka .J., Luckey H.A., Jonson W.A. Process for fabrication ON coatings for biomedical aplications. Sprey Conference. May 2-25, 1990; Long beac, CA: 433-438; Ducheyene P., Radin, S., Healy, K., Cuckler J.. The effect of plasma-spreying on the stractureand properties of calcium phospate ceramics. 34th Annual miting, Orthopedic Reasearch Sociaty. Februry 1-4, 1988, Atlanta, GA:50).

Another option is a method of coating the surface of metal implants with hydroxyapatite (HAP) (EN 2158189 C1, B05D 7/24, B05D 7/14, A61L 27/00, 27.10.2000), which involves mixing hydroxyapatite powder with a binder, drying, thermal annealing, and as a binder substance use phosphate ligament, and the powder (HAP) and phosphate bundle taken in a ratio of 1-1,5:1,5-2, calcination is carried out at varying between 250 and 600°C.

Metal implants, coated in a known manner, for use in medicine can improve clinical outcomes by regulating the ratio of amorphous and crystalline phase of hydroxyapatite and as a result to regulate the processes of bioactivity of the coating depending on the type and purpose of the implant.

However, the known method of obtaining a biomedical material is time-consuming, multistage, in the preparation of the coating material used is phosphoric acid, which imposes restrictions on the way in terms of ecology. The floor is uneven distribution of amorphous and crystalline phases, which affects the biological compatibility with living tissues.

The closest technical solution to the claimed invention is a method for orthopedic implant on onomatopeias alloy, comprising preparing an exothermic mixture of powders of cobalt oxide with the metal, which is used as: aluminum, magnesium, zirconium, compacting the mixture, initiating a chemical reaction self-propagating high temperature synthesis (SHS) by local heating of the compact under the pressure of the inert gas (helium, neon, argon, krypton, xenon, radon) 0.08 to 1.0 ATM and a temperature of preheating 933-950 To (660-677° (C)within 1-5 (US 6896846, SW 5/04, 24.05.2005). Additionally, the composition of the primary components for increasing the hardness of the final material can be introduced chromium, molybdenum, titanium, carbides, titanium nitride, and aluminum oxide or iron. The method has high productivity characteristic of all processes in the CBC mode. However, the resulting material has a complex composition, is a cast alloy (porosity less than 1%), does not have a sufficiently high bioactivity and compatibility with living tissues and ineffective for use in osteoplastic surgery.

The objective of the proposed solution is to create a new way to get a new biomedical porous material for implant-based alloy titanium-cobalt with desired structure and properties of high bioactivity, compatibility with living tissues and to the th can be used for bone implants in orthopedics and dentistry.

The technical result is a simplification of the production method, the improved biological compatibility of the obtained porous material based alloy titanium-cobalt with living tissues, with high mechanical characteristics.

The technical result is achieved in that a method of obtaining a biomedical material based alloy titanium-cobalt in the CBC mode, includes the preparation of an exothermic mixture of the raw materials of titanium powder and cobalt, adding to the mixture not more than 4 wt.% titanium hydride and not more than 15 wt.% amorphous nanosized hydroxyapatite powder or amorphous calcium hydroxyapatite nanocomposites nanocomposite calcium biopolymer natural origin, extrusion of a mixture of source powders billet, placing it in the reactor SVS, pre-heating the workpiece to a temperature 350-580°With the initiation of the combustion process in an inert atmosphere, followed by separation of the target product. As a biopolymer of natural origin are used, at least one selected from a range including: collagen, gelatin, keratin, sodium alginate, xanthan gum, carboxymethyl cellulose, chitosan.

Received biomedical material is a porous alloy corresponding to the formula TiCo, with a total porosity of 55-70%, with the share of open porosity 90-98%, with dimensions on the 200-800 μm, having an elongated shape, the walls between the pores also have a porous structure with a predominance of long round shape with a diameter of about 100 μm, the surface of the pore space of the alloy coated with compounds of calcium, phosphorus and oxygen, which are products of the decomposition of calcium hydroxyapatite, with a predominant content of perovskite calcium. The material of azithtomycin, the cells have a characteristic flatness and actively migrate inside the pore space.

Regarding the closest analogue of the proposed method is distinguished by the nature of the starting components, their ratio, the introduction of a mixture of different types of HAP calcium: amorphous, amorphous nanocomposites nanocomposite with biopolymer natural origin, titanium hydride and preheating the compressed mixture components.

Common symptoms are: local initiation of the combustion process in the pre-pressed mixture components in the atmosphere of inert gas, followed by reaction of the components in the CBC mode.

The input HAP powders, which were used for carrying out the invention:

- Hydroxyapatite calcium [CA10(PO4)6HE2] in the form of amorphous nanosized powder with particle sizes of 5-10 nm (TU 6-09-3538-91;

Sanitary-epidemiological conclusion No. p 26688.11.3 from 21.11.03. In brief recording gapamorf.

- Nanocomposite amorphous calcium hydroxylapatite with biopolymers (patent RU 2235061 C2, 27.08.2004), in particular "BULGIN" - an amorphous, nano-hydroxyapatite calcium in the form of microgranules with a diameter of 0.5 to 1 μm. Microspheres in turn consist of nanosized, bipolar spaced particles of calcium hydroxyapatite size of 5-10 nm and fibers, loosely formed of the same particles included in the organic shell of the biopolymer of natural origin. In brief recording gaporg.

- Calcium hydroxyapatite crystal, obtained by known methods, for example according to EN 2147290 C1, 10.04.2000 or EN 21492827 C1, 27.08.2004. In brief recording gapkr.

As titanium hydride was used micropowder obtained by the SHS method according to patent RU2208573, 20.07.2003. However, it can be used for any other powder, both domestic and imported.

Powders of titanium and cobalt used both domestic and foreign production: Ti (title, <25 μm) and With (<2 μm).

For testing of materials on cytotoxicity and determine their adhesive characteristics was used a primary culture of cells isolated from a human embryo for the period of 6 weeks. Cells were cultured in the medium of DMEM/199 (1:1) supplemented with 10% of embryonal the th calf serum (ETS) and 100 U/ml penicillin/streptomycin in an atmosphere of 5% CO 2. Culture cells at passage 11 (CD133-, Cd117-, CD45-, CD90+, CD54-, CD62L-, CD62P-, CD9+, CD34-, CD31-, CD71-, CD20-That CD157-, D106+, CD62E+) was used for materials testing, HLA typing of the culture says about the ownership of the cells to discharge mesenchymal stem. Cells were sown on the surface of samples with a density of 35 thousand/cm2and were cultured for 72 hours. Testing materials on the cytotoxicity was performed by examining the viability of mesenchymal stem cells on the surface of culture plastic (NUNC) with their co-cultivation with metals. The material considered toxic to cells when the culture more than 10% of apoptotic cells during co-cultivation with their study materials. Cell death was determined by the standard method to enable Trypanosoma blue. The adhesion characteristics of materials and their effects on the activity of the cells was determined by assessing the morphology of the cells seeded on the surface of the investigated materials. Evaluation of the morphology and viability of cells was performed on a microscope LUMAM I2 using the method of staining cells is 0.0002% solution of acridine orange in phosphate buffer. This dye selectively interacts with DNA and RNA chains is m intercalation or electrostatic attraction, respectively. When this is associated with a dye DNA fluorescent in the green light (525 nm), and electrostatically associated with RNA acridine orange fluorescent in the red range (>630 nm). This allows for an overall assessment of cell activity, proliferate, peace, apoptosis) (Darzynkiewicz Z. et al. Features of apoptotic cells measured by flow cytometry. Cytometry. 13(8):795-808, 1992).

The essence of the method is illustrated by examples.

Example 1 (comparative).

Prepare exothermic mixture of powders of titanium and cobalt, taken in stoichiometric ratio to obtain TiCo. Then the mixture was added 1 wt.% titanium hydride TiH2, 10 wt.% Gapkr., mix, pressed samples in the form of cylinders with a diameter of 12 mm, a height of 15-16 mm, with a density of 3.4-3.7 g/cm3place the samples in the SHS reactor, the reactor vacuum, filled with argon to a pressure of 1 ATM, samples are heated in the reactor to 580°With, then initiate the combustion process (reaction SHS) hot tungsten spiral. After completion of the reaction, the resulting material is cooled in argon, then extract and analyze known methods.

The obtained porous material is a sample with a total porosity of 50-55%, the percentage of open porosity is not more than 90% of the total porosity, the pore size ranges from 50 μm to 80 μm. The limit of compressive strength obtained is of material is 70 MPa.

Testing of this material on the cytotoxicity and the determination of its adhesive characteristics showed that the obtained porous material is toxic to cells and may not be used in medical practice.

Example 2.

Prepare exothermic mixture of powders of titanium and cobalt, taken in stoichiometric ratio to obtain TiCo. The mixture was added 1 wt.% titanium hydride TiH2mix , then in the mixture injected 10 wt.% Gaporg. and everything is thoroughly mixed. As biopolymer natural origin HAPorg. contains collagen. After pressing, the mixture samples in the form of cylinders with a diameter of 12 mm, a height of 13 to 14 mm, with a density of 3.0-3.2 g/cm3place in the SHS reactor, the reactor vacuum, filled with argon to a pressure of 1 ATM, then the sample is heated to 350°With, then initiate the process of burning hot tungsten spiral. After passing the SHS reaction, the resulting material is cooled in argon, extract and analyze known methods.

The obtained porous material is a sample with a porosity of 55-60%, the percentage of open porosity is 90-98% of the total porosity. The pores have an elongated shape, the pore size of 200-500 μm. While walls between pores also have a porous structure, dominated by pores rounded, with a diameter of priblizitel is but 100 microns.

The limit of the compressive strength of the material obtained is 130 MPa. The study of the microstructure and elemental composition of the synthesized composite materials showed that the matrix of the samples consist of one phase TiCo. The surface is then covered with compounds of calcium, phosphorus and oxygen, which are products of decomposition of the gap, with a predominant content of perovskite calcium formula CaTiO3.

Testing of this material on the cytotoxicity and the determination of its adhesive characteristics showed that the obtained porous material azithtomycin, has a high activity of mesenchymal stem cells showed characteristic raspletanie and migration of cells inside the pore space, RNA synthesis) and is characterized by high integration potential.

When using gaporg. with other biopolymers specified in the formula, get porous material with the properties of the material according to example 2 and high biomedical characteristics.

Example 3.

Prepare exothermic mixture of powders of titanium and cobalt, taken in stoichiometric ratio to obtain TiCo. The mixture was added 4 wt.% titanium hydride TiH2mix , then this mixture was added 15 wt.%. Gapamorf., all are thoroughly mixed. Pressed samples in the form of cylinders with a diameter of 12 is m, the height of 12-14 mm, with a density of 3.2-3.5 g/cm3place the samples in the SHS reactor, the reactor vacuum, filled with argon to a pressure of 1 ATM, and then placed the samples heated up to 580°and initiate the process of burning hot tungsten spiral. After passing the SHS reaction, the resulting material is cooled in argon, extract and analyze known methods.

The obtained porous material is a sample with a porosity of 60-70%, the percentage of open porosity is 95-98% of the total porosity. The pores have an elongated shape, the pore size of 300-800 μm. While walls between pores also have a porous structure, dominated by pores rounded, with a diameter of approximately 100 μm. The limit of the compressive strength of the material obtained is 35 MPa. The study of the microstructure and elemental composition of the synthesized composite materials showed that the matrix of the samples consist of one phase TiCo. The surface is then covered with compounds of calcium, phosphorus and oxygen, which are products of decomposition of the gap.

Testing of this material on the cytotoxicity and the determination of its adhesive characteristics showed that the obtained porous material azithtomycin, has a high activity of mesenchymal stem cells, characterized by a higher integration is otentials compared with the material, obtained in example 2.

The study of the structure of the obtained materials was conducted on the samples by the method of local x-ray analysis x-ray microanalyzer "Superprobe JCXA-733". The strength characteristics of the materials were determined under uniaxial loading samples on the test car "Instron-1195" (on compression). The phase composition of the obtained materials was determined by the method of x-ray phase analysis by setting a DRON-3 copper radiation.

Assessment of the toxicity of composite materials Ti-Co-HAP obtained by the SHS method, was performed using scanning electron microscopy (SEM) according to the criteria: cell adhesion and raspletanie cells. This used the standard test for cytotoxicity of materials using culture of embryonic skin and muscle" in human fibroblasts.

Figure 1 shows pictures of the patterns of samples of materials obtained by the proposed method. All samples have a uniform open porosity of 50-70%. It is seen that depending on the chemical and phase composition gap, added to the initial mixture, changing not only the magnitude of the porosity, but also the shape and size of the pores of the material obtained. So when using crystalline HAP (comparative) the pore size of 50-80 microns, dominated by pores rounded (figa). With the addition of amorphous gap and is of composite amorphous calcium hydroxylapatite with biopolymers porous structure of the samples is changed - the pores have an elongated shape, pore size 200-800 μm (figb, 1B). While walls between pores also have a porous structure, dominated by pores rounded, with a diameter of about 100 μm. The sample containing no additives gap (comparative), is characterized by an uneven porosity and pore size ranging from 50 μm to 80 μm (high).

The study of the microstructure and elemental compositions of the samples of the same composition, as in figure 1, showed that with the addition of the gap in the initial mixture matrix synthesized samples consist of one phase TiCo (figure 2). The surface of the pores of the samples with additives gap is covered by compounds of calcium, phosphorus and oxygen. Matrix samples synthesized without additives gap consists of two phases - TiCo and TiCo2(Figg comparative).

Thus, the introduction of gapamorf. and gaporg. in the original mix has a positive effect on the performance of the new material (fraction of open porosity 90-98% and the tensile strength of the material in compression up to 130 MPa).

In addition to these characteristics of the new material shows a high level of biological compatibility with the cellular elements on which is based the predicted their ability to osseointegration.

A study using SEM showed that the method based on the study of the contact of cultured embryonic fibroblasts man is and with the sample surface, allows to investigate not only the Toxicological characteristics of the test samples, but their integration potential with respect to living tissue, which were the indicators used in the adhesion of cells and their rasplastyvanija.

Figure 3 presents the growth of mesenchymal stem cells in the pore volume and on the surface of the synthesized samples: a) example 1 b) example 2) example 3, g) Ti+Co (comparative).

Figure 4 presents the growth of cellular tissue in the pores of the samples a) sample 1; b) case 2; C) according to example 3; g) Ti+Co (comparative).

It is established that the high integration potential samples obtained from Ti+From+10%gaporg.+1%TiH2(example 2) and Ti+Co+15%gapAmort.+4%TiH2(example 3). Using gaporgnot only facilitates the formation of pores, but also leads to modification of the properties of their surface. According to the results of the elemental analysis of the surface pores contains calcium, phosphorus, oxygen, which is more favorable for the cell.

The obtained material showed that the cells have a characteristic flatness and actively migrate inside the pore space. The use of a fluorescent dye, which selectively interacts with DNA and RNA by intercalation or electrostatic attraction, respectively, has allowed to establish proliferative and biosynthetic activity of the cells during culturing them on the surface of this material. This cell death is not more than 5%, a rate similar to that in the control when growing cells on the surface specially treated for cell growth polystyrene culture plastic manufactured by NUNC).

Thus: a new way to generate new porous biomedical materials based on alloy Ti-Co in the CBC mode, allowing the formation of materials with a uniform single-phase structure and properties of high bioactivity and compatibility with living tissues; the optimal parameters of the method.

The claimed combination of features of the process allows to obtain a new biomedical material for implants with an open porosity of 55-70%, and the surface structure of the pore space, allowing the adhesion and migration of mesenchymal stem cells.

The new material can be used for bone implants in orthopedics and dentistry.

Testing of the material on the cytotoxicity and the determination of its adhesive characteristics showed that the obtained porous material has no toxicity, has a high activity of mesenchymal stem cells and high integration potential.

1. A method of obtaining a biomedical material based alloy titanium-cobalt in the CBC mode, the switch is in store cooking exothermic mixture of the raw materials of titanium powder and cobalt, add in the mixture is not more than 4 wt.% titanium hydride, and not more than 15 wt.% amorphous nanosized hydroxyapatite powder or amorphous calcium hydroxyapatite nanocomposites nanocomposite calcium biopolymer natural origin, extrusion of a mixture of source powders billet, placing it in the reactor SVS, pre-heating the workpiece to a temperature 350-580°With the initiation of the combustion process in an inert atmosphere, followed by separation of the target product.

2. The method according to claim 1, characterized in that the biopolymer of natural origin are used, at least one selected from a range including: collagen, gelatin, keratin, sodium alginate, xanthan gum, carboxymethylcellulose, chitosan.

3. The material obtained by the method according to claims 1 and 2 characterized by the fact that he is a porous alloy corresponding to the formula TiCo, with a total porosity of 55-70%, with the share of open porosity 90-98%, with a pore size of 200-800 μm, having an elongated shape, the walls between the pores also have a porous structure with a predominance of long round shape with a diameter of about 100 μm, the surface of the pore space of the alloy coated with compounds of calcium, phosphorus and oxygen, which are products of the decomposition of calcium hydroxyapatite, with a predominant content of perovskite calcium.

4. The material according to claim 3, the characteristic is kersulis fact, what he azithtomycin, the cells have a characteristic flatness and actively migrate inside the pore space.



 

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3 dwg, 1 ex

FIELD: medicine, operative gynecology.

SUBSTANCE: one should cut and remove a triangular fragment out of posterior vaginal wall, against the top of separated triangle it is necessary to make a longitudinal median incision of vaginal wall up to posterior arch, then one should conduct maximal separation of posterior vaginal wall laterally to the walls of small pelvis along opening overstretched anterior rectal wall covered with rectovaginal fascia up to lateral walls, with either one or two purse-string sutures applied onto rectovaginal fascia at capturing anterior rectal wall. One should reconstruct a muscular-fascial rectal sheath during the whole length of rectovaginal septum with separate more surface sutures onto rectovaginal fascia with adjacent muscular elements. Reconstructed muscular-fascial sheath should be sutured with super-elastic ligature out of titanium nickelide. Moreover, sutures should be applied at stitch length of about 5-6 mm and distance between sutures being 5-6 mm in longitudinal and cross-sectional directions by netting-type against external rectal sphincter up to posterior vaginal arch and lateral rectal walls. One should isolate and suture muscles lifting anus and muscular-fascial perineal foundation. Then it is necessary to suture the edges of vaginal wound, subcutaneous layer and perineal skin. The innovation in question enables to reconstruct normal anatomo-topographic position of anterior rectal wall and posterior vaginal wall.

EFFECT: higher efficiency of reconstruction.

8 dwg, 2 ex

FIELD: medicine.

SUBSTANCE: the present innovation deals with a medicinal prosthesis that contains metallic material, such as titanium or its alloy, in which surface parts of metallic material area covered with the layer of the corresponding hydroxide material, such as titanium hydroxide. Preferably, hydroxide layer contains one or more biomolecular substances being connected with it. Also, the innovation in question refers to electrolytic process for obtaining a medicinal prosthesis. Metallic prostheses are of improved biological compatibility.

EFFECT: higher efficiency.

24 cl, 8 ex, 3 tbl

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