Method of treating periodontitis

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely surgical stomatology and can be used for treating paradontium. Periodontal osteogenesis and neogenesis are ensured by removing pathologically changed structures and introducing the mesenchymal stem cells suspended in a carrier into the defect cavity. As the stem cells, the mesenchymal cord and placental stem cells received after easy delivery are used. The cells 5-10·106 are injected in 1-2 ml of patient's blood plasma.

EFFECT: method provides more effective replacement of the manifested periodontal defect associated with paradontium, reduced length of treatment with no side effects.

3 ex

 

The invention relates to medicine and can be used in dental surgery in the treatment of diseases periodontitis.

Periodontal disease is spreading among the population of the 4-th place in the list of non-communicable diseases, and the age of the patients does not play a role: for 30 years the disease register 50% of the population.

In the development of periodontitis involved exogenous and endogenous factors such as the composition and properties of saliva, microorganisms, Tartar formation, etc. of the Immune and hormonal factors are key in triggering the mechanism of development of severe forms of inflammatory-destructive periodontal disease. Periodontitis is characterized by the fact that in patients affected all parts of the periodontium with bone resorption of the alveolar ridge, resulting in the death of the restraint apparatus of teeth and loss of teeth. By treatment of periodontitis, it is necessary to accelerate the processes neoangel and osteogenesis and repair all tissues affected by disease.

Known way to restore periodontal tissues in the defect with osteoplastic material "Cerasorb", with platelet enriched plasma (Parodontologie, 2004, №3(32). The disadvantage of this method is that the osteoconductive material is saturated with platelet plasma. Because of the availability of the only elements of the blood, regeneration of periodontal tissues is slow.

Known method periodontal treatment using collagen sponge "collegit" and postnatal human fibroblasts directly in the field of periodontal defect (Parodontologie, 2004, №3(32)), material used - collagen-chitosan gel. The disadvantage of this method is the use of postnatal fibroblasts and the use of multicomponent systems in the gel, and the manufacturing time of the gel from 1 to 3 days.

Modern and actively developed a method for the treatment of periodontal defects is treatment using stem cells, in particular mesenchymal stem cells.

There is a method of treatment of inflammatory periodontal diseases with the use of compositions based on cell cultures containing the fibroblast cultures, growing environment, osteoplastic material (U.S. Pat. Of the Russian Federation No. 2210352). As of fibroblast culture it contains a strain of diploid lung cells of a human embryo LAC-4(81), as the medium growth medium NEEDLE-MEM with the addition of 10%serum fruits cattle, as osteoplastic material Hapka or Callahan. When using the known method possible immune reaction in the patient. And the most important clinical and histological results after such interventions are highly ambiguous: is formed defunct the national scar, celebrated the expansion of the boundary of the epithelium and the absence of normal periodontal attachment in the area of healing of periodontal tissues, in the substituted section of the bone is marked excess calcification with impaired skeletal tissue functions, often observed reaction to the foreign body until the pyrogenic effects. There is a risk of human transmission of infections in animals.

There is a method of treatment with mesenchymal stem cells (MSCS) (U.S. Pat. Of the Russian Federation No. 2265445). In this way, the suspension of MSC or chondroprotective cells or combinations thereof is administered intraligamentary with the lateral and medial sides of each affected caries of the tooth and/or subperiosteal in periodontal sites of the upper and lower jaws. Mesenchymal stem cells by this method is obtained from fetal donor material or from the recipient's own tissue (for example, from bone marrow, adipose tissue). As the cell material used alien chondroprotective cells, sometimes in combination with alien MSC. Made of MSC slowly and not fully acquire functional properties of osteocytes. The treatment period of 6 months, and orthopantogram significantly increase the optical density of the bone of the alveolar process compared to the "before treatment" 49±3%, p=0.05. In comparison with the known SPO is obom autologous MSCS have advantages, because eliminate the immune response, blood-brain reaction. In the combined treatment as cellular composition, and enter the volume of the cell suspension.

The closest method of treatment of periodontitis is the way to restore the bone of the alveolar ridge of the jaw bone and periodontal tissues with reduced capacity, including the removal of pathologically altered structures, the filling of the defect with a blood clot from the jaw bone and/or periodontal ligament. In the area of tissue defect transplanted autologous vascular cell fraction containing mesenchymal stem cells - fibroblast-like cells isolated from adipose tissue and purified from Mature adipocytes, without prior cultivation, and when the recession of periodontal tissues exposed root surfaces of the teeth previously covered palatal podperevalny connective tissue flap (U.S. Pat. RF 2320285). When using this method cannot eliminate the risk of neoplastic processes and reduce the time of treatment.

The objective of the present invention is to improve the efficiency of recovery expressed periodontal defect associated with periodontal disease, the reduction of treatment time.

The essence of the invention is a method for restoring the Oia bone and periodontal tissues, including the removal of pathologically altered structures and introduction to defective cavity mesenchymal stem cells suspended in the medium. As stem cells using mesenchymal stem cells from umbilical cord or placenta embryo obtained after a normal birth, with carry introduction cells by injection of 1-2 ml of blood plasma of the patient containing 5-10·106cells.

Example 1. Obtaining mesenchymal stem cells

Mesenchymal stem cells of the umbilical cord or placenta were obtained in accordance with the standard method as follows. Samples of the umbilical cord or placenta were collected after normal delivery at 39-40 weeks of gestation. Vein of the umbilical cord and placenta were Coulibaly and washed first with Hanks solution, and then 0.1% solution of collagenase type I, prepared in DMEM without serum, and incubated at 37°C for 30 minutes and Then the vessels were again washed with Hanks solution, the fabric was subjected to a short soft mechanical stress and collecting the separated cells. The obtained cells were besieged by centrifugation. Sediment resuspendable in DMEM containing 10% serum, 100 units/ml penicillin, 100 units/ml streptomycin, 2 mm glutamine, 1 mm sodium pyruvate, 10 ng/ml basic fibroblast growth factor (β-FGF), was transferred into the culture Cup and cultivar the Wali before the formation of the monolayer, changing environment 2 times a week. When reaching the monolayer cells were perseval in the ratio of 1:3. In the case of cell culture in clinical trials of the method after the last subculture under cultivation instead of fetal calf serum used the serum obtained from the patient's own blood.

Example 2. Laboratory tests on animals.

The method is tested in experiment 16 rats in the clinic for 7 patients. Of male rats weighing 300-350 g were subjected to light anesthesia. Periodontitis caused by the imposition of a pressure ligatures of the nylon thread around the neck of the first lower right molar. 16 rats 7 days after applying ligatures in the gum was introduced 500 thousand cells in sterile saline R-re. 8 control animals in the gums were injected with physiological R.-R. Even after 2 weeks, all animals were killed and subjected to the study of the periodontium and alveolar bone in the area 1 lower left molar. Control animals were observed pronounced inflammation of the periodontium and the signs of destruction of the alveolar bone, as well as increased activity of metalloproteinases. 14 experimental animals, these effects were completely absent, and the two were poorly marked.

Experiment on animals showed a significant acceleration of the process of primary wound healing when using mesenchyme is lnyh stem cells of the umbilical cord or placenta embryo received after normal delivery.

Example 3. In clinical trials.

The patient is complaining of tooth mobility. According to the examination 2-3 degree, bleeding gums. On the basis of clinical and rentgenologicheski examination diagnosed with periodontitis severe atrophy more than 1 of the alveolar ridge. The patient held: professional hygiene of the oral cavity, the teeth of the upper and lower jaw siniawan selectively thorough curettage of pathological pockets. After the above mentioned treatment on the basis of voluntary informed consent of the patient conducted substitution of cell therapy in accordance with the claimed method of treatment using injections of a patient's blood.

When using the inventive method in clinical trials were also obtained positive results. Introduction cells was performed by injection of 1-2 ml of blood plasma of the patient containing 5-10·106cells. All 7 patients up to 5 days decreased periodontal pockets, disappeared breath, stopped bleeding gums, mucous membrane bought a pale pink color, i.e. was visualized positive dynamics of the disease. When rentgenologicheski control after 6 months was determined by the growth of bone tissue, reducing the depth and width of the periodontal karmano is, what is the evidence of remineralization of bone tissue. Tooth mobility was gone.

The use of stem cells mesenchymal stem cells of the umbilical cord or placenta embryo received after normal delivery, helped to increase the effectiveness of treatment, to increase the reproducibility of results and to avoid unwanted effects.

Method of recovering bone and periodontal tissues, including the removal of pathologically altered structures and introduction to defective cavity mesenchymal stem cells suspended in the medium, characterized in that as stem cells using mesenchymal stem cells from umbilical cord or placenta embryo obtained after a normal birth, with carry introduction cells by injection of 1-2 ml of blood plasma of the patient containing 5-10·106cells.



 

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2 tbl, 1 ex

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2 tbl

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1 tbl, 3 dwg

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