Composite ceramic bone grafts made of ceramic material of zirconium oxide - aluminium oxide system

FIELD: medicine.

SUBSTANCE: invention describes a composite ceramic bone graft containing a porous ceramic carrier made of zirconium oxide - aluminium oxide; the carrier is coated with a layer of hydroxyl apatite and platelet-rich plasma; the carrier is made by preparing a mixture ceramic powder and foaming agent Al(OH)3 or Zr(OH)4, adding distilled water to give the moulding properties and caking the end product.

EFFECT: composite ceramic bone graft of the ceramic material of zirconium oxide system is effective and applicable in medicine for synthesis of the anatomic continuity and functions of the bone tissue.

1 ex, 14 dwg

 

The invention relates to medicine, namely to traumatology, orthopedics, regenerative medicine, dentistry and maxillofacial surgery to restore the structure and function of bone tissue.

The actual problem in medicine is the restoration of the anatomical integrity and function of bone tissue. One of the reasons for the disability population are diseases and post-traumatic conditions, coupled with the need for reconstruction and replacement of sections of bone tissue, prosthetic fragments of the musculoskeletal system. Bone defects were formed after injuries, surgeries, tumors, require a full plastic materials. Duration of treatment patients and their disability ranges from 6-8 months to 2-3 years, and 6-8% of cases end in a permanent disability that entails significant economic costs and social significance. Hence the importance of early research management capabilities reparative processes. The timing adjustment through an adequate choice of materials or the creation and use of materials with desired set of properties.

Currently, medicine is known for a wide range of upper-matrices. The greatest interest for tissue engineering represent a mule is thepotential stromal cells in the bone marrow (MSC), capable of differentiation into osteogenic, chondrogen and adipokines directions [1]. Introduction cells leads to the replacement of lost cellular elements, induction of regeneration by releasing growth factors and differentiation, formation of extracellular matrix, which greatly speeds up the recovery of damaged tissue [2]. Immobilization of cultured cells on the matrix carrier provides the highest concentration of cells in the damaged area, differentiation in a given direction and increases the survival rate of the cells during transplantation.

Demineralized bone matrix (DCM) due to natural architectonics, osteogenic potential, due to the presence of biologically active substances, is one of the most popular materials for osteoplasty. Shows the effectiveness of media-based DCM with immobilized MSC recovery defects of the parietal bones in rats and dogs [3]. Patented technology for the treatment of false joints using biotransplant based DCM and autologous MSCS [4]. The disadvantages of this xenogenic material are the need for handling aggressive fluids (reduce cell adhesion), potential immunogenicity and the possibility of transfer of infectious diseases.

Shiro is used on synthetic biodegradable carriers on the basis of polylactic and polyglycolic acids and their copolymers in combination with MSC also demonstrate the induction of osteogenesis in vitro and formation of bone tissue through 6-9 weeks in experiments on animals [5]. Membrane of polylactide and/or polyglycolides, impregnated with chondrocytes or bone marrow MSC, patented as a method of treatment of bone and cartilage defects in humans and animals [6]. The disadvantages of such media is quick and not always predictable degradation of the matrix, which is accompanied by a local decrease of pH in the field of implant that reduces the effectiveness of their use [7].

Known biotransplant based on highly porous ceramic material system Zirconia - alumina and multipotent stromal cells of human bone marrow to restore long bone defects (patent RF № 2386453, 2008) adopted for the prototype, characterized in that it contains autologous or donor multipotential mesenchymal stromal cells (MSC) from the bone marrow and the media created on the basis of a highly porous ceramic material system Zirconia - alumina technology duplication polyurethane basis, with the media densely seeded MSC, cultured from 1 to 3 passages, at the same time on the same carrier immobilized from 200 to 500 thousand cells, the vitality of which is at least 90%, and functional orientation is confirmed by the ability of the directed differentiation of IU is dermaline line and demonstrates the expression of stromal markers CD 90, CD 105, 60-90% of the cells and the absence of expression of the marker CD 34.

However, working with multipotent mesenchymal stromal cells (MMSC) is very laborious and requires a considerable period of time from 2-3 weeks up to 1 month from the period of sampling from the patient prior to implantation. In addition, when the number of passages in cell culture, an increasing number of mutations.

Disclosure of the invention.

The technical result of the invention is the creation of a composite bone-ceramic implant-based ceramic material system Zirconia - alumina, which optimizes the process of repair and restoration of damaged bone structure. Increases biological compatibility, strength properties. Ensures the absence of immune response. Enhances the reparative processes in the damaged tissues. Provides fast healing and restoration of bone structure.

This technical result is achieved in that the composite bone-ceramic implant containing porous ceramic media based on Zirconia - alumina in the carrier coated with the layer of hydroxyapatite and platelet-rich plasma, while the carrier is obtained by preparing a mixture of ceramic powder based on ZrO2(Mg, Y) or Al2About3and additives of plastipak the ora and pore-forming Al(OH) 3or Zr(OH)4followed by adding distilled water to give a mixture of molding properties and sintering the finished product.

The invention is illustrated in the following graphic materials, where Fig.1 - General view of the product.

A variant embodiment of the invention.

Step 1. The manufacture of porous ceramic material. The manufacture of porous ceramic material is carried out in a known manner by the RF patent № 2476406, 2008, by preparing a mixture of ceramic powder and an additive acting as a plasticizer and blowing agent, forming the desired product configuration and subsequent sintering, as the ceramic powder uses ultra-fine powder of solid solutions on the basis of ZrO2(Mg, Y) or ultra-fine powder of Al2O3and as a plasticizer and blowing agent use the Hydrosol Al(OH)3or Zr(OH)4in an amount of from 1 to 50% by volume of the mixture, add, stirring, distilled water to give a mixture of molding properties, then hold the molding by pressing under a pressure of from 12 to 25 kN, and sintering the product is carried out at a temperature 1450-1600°C.

Step 2. Manufacturer WPC - sawdust further biological hydroxyapatite. Made of biological hydroxyapatite in a known manner by the RF patent№ 2232585, 2001 by cleaning the bones, sawing into fragments, washing with water and deproteinization at 37°C with 0.01% solution himopsina within 72 h and 10% hydrogen peroxide solution for 48 h, the processing liquid ether, 10% solution of lithium chloride for 16 h and washing with sterile distilled water, deproteinizing bone after washing with sterile distilled water crushed to particle size of 0.1-0.5 mm, obtained deproteinizing bone meal are combined at a temperature of 38-40°C with 9% gelatin, prepared on preserving solution with the addition of antibiotic, preserved at -20°C and sterilized.

Step 3. The coating of biological hydroxyapatite. On the composite bone-ceramic implants is a cold gas-dynamic spraying of biological hydroxyapatite by installing a cold gas spraying (CS) with the following parameters: working gas is air; the pressure in the prechamber 0.3 to 5 MPa; the temperature in the prechamber 0-1000°C; gas Flow rate ≤1.5 m3/min; power consumption ≤20 kW.

Step 4. Application of platelet-rich plasma on the composite bone-ceramic implant with a coating of biological hydroxyapatite. 20-30 minutes before operations are performed blood sampling 10-15 ml of the patient in a vacuum tube. The tube is blood centrifuged for medical centrifuge at 1500 rpm for 15 minutes. In the centrifuge process is the separation of blood into fractions with the formation of the median fraction on the border between erythrocytes and plasma. The resulting platelets are extracted with a syringe in a sterile Petri dish, where directly immerse the implant.

Experimental study. Implantation of experimental samples of materials were carried out in the compact bone defect Mature rats male Wistar rats at the age of 5-6 months, weighing 260-320 grams. Model perforating defect of the mandible with the subsequent filling of the defect with the materials used for the study of biological response for the experimental sample.

Composite bone-ceramic implants based on Al2About3and ZrO2-ceramic, cylindrical shape with a height of 2-3 mm, diameter 2 mm and coated with a biological hydroxamate. To stimulate bone formation was used platelet-rich plasma (PRP). Animals were taken from the experiment on the 15th and 30th day. Thus were formed the following groups/

In the first series - studied biological response upon implantation in the compact bone of experimental samples of bone-ceramic implants, i.e., Zirconia ceramics with surface coating deproteinisation holocaustum powder. Number of animals - 6, to icesto operations - 12. The follow-up period was 15 and 30 days after implantation.

In the second series studied biological response upon implantation in compact bone of the experimental modified samples of bone-ceramic implants, i.e., samples modified platelet-rich plasma. Number of animals - 6, number of operations 12. The follow-up period was 15 and 30 days after implantation.

Experimental studies were performed according to the elaborated methods and protocols "NIETO them. J. L. Ziviani" Ministry Of Health Of Russia.

The results of the first series of experiments, after 15 and 30 days after implantation.

1.1. Composite bone-ceramic implants in 15 days after implantation. In Fig.2 presents a composite bone-ceramic implant in the perforation defect of the mandible in 15 days after implantation. White arrows indicate the bone tissue on the surface of the sample, black - bridges between the edge of the bone defect and the newly formed bone. In Fig.3 presents extracted from the bones of an animal bone-ceramic implant. On the surface of the bone-ceramic implant - the newly formed bone tissue, painted in the color purple. Quite a large array of newly formed bone with formation of young bone beams.

1.2. Composition to the IDT-ceramic implant 30 days after implantation. Composite bone-ceramic implant 30 days after implantation is shown in Fig.3, Fig.4, Fig.5. In Fig.4 presents the appearance of a composite bone-ceramic implant is implanted in the lower jaw of the experimental animal. Observed the formation of newly formed bone at the periphery of the implant and on its surface. In Fig.5 presents the appearance of a composite bone-ceramic implant extracted from the jaw bone of an experimental animal. Himself composite bone-ceramic implant presents white. On its surface are dark blotches corresponding newly formed bone on the porous surface of the implant. In Fig.6 shows the appearance of the bone defect after removal of her implant. Edge of the bone defect rather rounded, correspond to the shape of the implanted composite bone-ceramic implant. Sometimes small bumps corresponding to the forming bone "bridges" to the pores of the composite bone-ceramic implant. In the lower part there is a configuration change of bone defects due to formation of surface layers on the implant, directed from the periphery to the center of the formed bone defect.

The results of the second series of the experiments after 15 and 30 days after implantation.

2.1. Composite bone-ceramic implant with a modified enriched vombatidae plasma within 15 days after implantation. The appearance of the composite bone-ceramic implant with a modified platelet-rich plasma, after 15 days after implantation into the bone of the lower jaw of the experimental animal (rat) looks as shown in Fig.7 and Fig.8. Some sites of bone resorption, places, sites of deposition of the newly formed bone on the composite bone-ceramic implant with a modified platelet-rich plasma. Strata to reach from the bones of the lower jaw to the center of the implant. In Fig.9 and Fig.10 presents extracted from the bones of the lower jaw composite bone-ceramic implant with a modified platelet-rich plasma. There are clear signs of biological compatibility of the implant, which is expressed in the ability to the formation on the surface of young bone, dyed in purple color when counterstaining with hematoxylin and eosin

2.2. Composite bone-ceramic implants with a modified platelet-rich plasma after 30 days after implantation. The appearance of the composite bone-ceramic implant with a modified about kamennoi platelet plasma after implantation into the bone of the lower jaw of the experimental animal (rat) looks like this, as is shown in Fig.11 and Fig.12. Newly formed bone was almost completely covers the surface of the implant. Strata to reach from the bones of the lower jaw to the center of the implant. Composite bone-ceramic implant with a modified platelet-rich plasma, extracted from the mandible experimental animal after 30 days after implantation is shown in Fig.13 and Fig.14. Newly formed bone is formed directly on the implant surface. A large array of fabrics dyed represents the deposition of the newly formed bone. Observed the formation of osteoid on the implant surface and the area of the bone-implant. The formation comes from the bone bed to the center of the implant. On the periphery of the implant structure of bone is identical to the intact bone of the jaw, as the distance from the bone bed structure of the newly formed bone unstructured, which is typical for the stage of formation of osteoid.

Thus, the results of experimental studies have shown that the inflammatory reaction caused by the implanted materials in any series of the experiment, was not observed. Bone formation is by layering, in some areas, the mass of newly formed bone on the implant is large, but always there is a connection with the mater is nskoi bone. Proliferation and migration of bone cells leads to appositional bone growth on the implant surface. In all periods of the experiment, there is a pronounced mineralization of the extracellular matrix calcium ions. When applying the modified platelet-rich plasma is the activation of both stromal and hematopoietic component of bone formation (developing the stroma of the bone marrow and are hematopoietic cells). In late period (30 days) use of a modified platelet-rich plasma to remove the implant from the bone of the jaw was not possible due to immured implant newly formed bone tissue. In late period (30 days) is showing signs of biodegradation of the material of the implant, more pronounced in the case of the use of platelet-rich plasma. Resorption of the material of the implant occurs without apparent involvement of histiocytes and osteoclasts.

The study results of the claimed invention demonstrate the ability to effectively use in medical practice for repair of damaged bone structure as a composite bone-ceramic implant-based ceramic material system Zirconia - alumina optimizes the process of repair and recovery, damaged the th bone structure, fast healing and restoration of bone structure. Increases biocompatibility and mechanical properties, ensures the absence of immune response, enhances the reparative processes in the damaged tissues.

Research results indicate that composite bone-ceramic implant-based ceramic material system Zirconia - alumina, effective and useful in medicine to restore anatomic integrity and function of bone tissue.

The invention can be manufactured using hi-tech equipment, modern technologies and materials.

Bibliography

1. Pittenger, M. F., Mosca, J. D., McIntosh K. R., Human mesenhymal stem cells: progenitor cells for cartilage, bone, fat and stroma. Curr Top Environ Immunol. 2000; 251:3-11.

2. Salgado A. J., Oliveira J. T., Pedro A. J., R. L. Reis Adult stem cells in bone and cartilage tissue engineering. Curr. Stem Cell. Res. http://Ther.2006.sep; 1(3):345-64.

3. Liu W., Cui L., Cao Y. Mesenhymal stem cells and tissue engineering. Methods in enzimology. 2006. Vol. 420. P. 339-361.

4. RF patent № 2309756 from 10.11.2007.

5. Yao J., Radin, S., Leboy p, Ducheyne P. The effect of bioactive glass content on synthesis and bioactivity of composite poly (lactic-co-glycolic acid)/ bioactive glass substrate for tissue engineering. Biomaterials. 2005 May; 26(14):1935-43.

6. Bostman O., Paivarinta U., Partio e, Vasenius j, Manninen m, Rokkanen P. Degradation and tissue replacement of an absorbable polyglycolide screwin the fixation of rabbit femoral osteotomies. J. Bone Joint. Surg. Am. 1992. Aug; 74(4): 1021-31.

7. RF application No. 2000109319 from 20.02.2003.

HDMI is traditional bone-ceramic implant, containing porous ceramic media based on Zirconia - alumina, characterized in that the carrier is coated with the layer of hydroxyapatite and platelet-rich plasma, while the carrier is obtained by preparing a mixture of ceramic powder based on ZrO2(Mg, Y) or Al2About3and additives plasticizer and pore-forming Al(OH)3or Zr(OH)4followed by adding distilled water to give a mixture of molding properties and sintering the finished product.



 

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