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discussion

Defects of an osteal tissue, its age loss or loss in connection with pathological conditions cannot be completely eliminated by physiological neogenesis or by means of a surgical intervention.

In such situations biomaterials or their synthetic analogues are used.

Available methods far not always allow not only to fill volume of available defect, but also to frame conditions of performance of mechanical function of a bone. Differently, we have come nearer to creation of sufficient volume of an osteal tissue, but structural characteristics received osteal regenerata are far from the wished. We also have tried to solve the arisen problem in the presented research.

The method influencing mechanisms remodelirovanija of an osteal tissue, is bound to use rekombinantnogo human morphogenetic fiber of a bone (rhBMP-2) which, in turn, activates function of osteoblasts. Studying of properties and use possibilities rhBMP-2 is spent now in the majority mediko - the biological centres of the world.

At the first investigation phase the problem consisted in synthesis and reception is immediate the fiber for planned researches. This problem has been successfully solved in commonwealth with Institute of theoretical and experimental biophysics of the Russian Academy of Sciences, laboratory of fabric engineering, the principal of laboratory of d.f. TH n Akatov Vladimir Semenovich.

Fiber has been received by a method of gene engineering which included following stages:

1. Gennoinzhenernaja assemblage of a biological design;

2. Introduction of the framed molecule in cells of bacterium E.Coli;

3. An operating time of sufficient volume of bacteriemic mass for reception of necessary quantity of fiber;

4. Allocation of fiber from bacteriemic mass and its biochemical clearing.

The carried out analysis of literary data has allowed to choose optimum a biomaterial necessary for sharing with rhBMP-2. As such material the representing sterile bioplastic material on the basis of osteal ksenokollagena, sated sulfatirovannymi glikozaminoglikanami (sgag) for the referred osteal neogenesis, the manufacturer of Joint-Stock Company «EOZ VladMiVa» has acted "osteodent",

Further, having received a necessary composition (a biomaterial with rhBMP-2), we have started laboratory researches.

In laboratory stromalnoj regulation of immunity FGBU scientific research institute of N.F.Gamalei under a management d.m.n. Prof. R.K.Chajlahjana has been planned and successfully made the experiment which purpose put revealing available mitotic activity of cells-predecessors stromalnyh fibroblasts of an osteal brain and visually to estimate their adhesive activity on a surface of the offered biomaterial containing rhBMP-2.

Cells-precursors stromalnyh fibroblasts of an osteal brain received a following method: rats lulled an Aether. With observance of rules of an asepsis through a cut on a back part of a hip allocated femoral and tibial bones. Epiphysises of bones cut off and shpritsom blew an osteal brain in a vial with a nutrient medium. Fragments of an osteal brain passed through a syringe with consistently decreasing diameter of needles at the minimum pressure in it before reception of a homogeneous suspension of cells. A suspension washed a centrifugation at 4°С (400g), a deposit resuspendirovali in a fresh nutrient medium twice, filtered through 4 layer kapron filter.

For reception of strains stromalnyh cells-precursors of an osteal brain eksplantirovali in vials. For 10-14 day kultivirovanija, when

Colonies stromalnyh fibroblasts have been completely generated, spent I passage.

For studying of interaction of strains of osteogene cells - of precursors with the osteoplastic material sated rekombinantnym with human morphogenetic fiber of a bone rhBMP-2, it sowed passivirovannymi cells. kultivirovanie spent within 5 7 days. Then made bracing.

Before bracing kulturalnuju medium merged, a material some times washed out a normal saline solution and for 30 minutes filled in 80 % with ethanol. Fixed on the osteoplastic material containing rekombinantnyj morphogenetic fiber of a bone rhBMP-2, cultures painted on Gimze. After that estimated the received result under a microscope.

The visual estimation of cells-predecessors on an osteoplastic material by means of a scanning supermicroscope has shown, that presence rhBMP-2 does not interfere with an attachment and bracing of a cellular material on nedemineralizovannom an osteal matrix. The given research was spent to laboratories of anatomy of microorganisms FGBU scientific research institute of N.F.Gamalei, the principal of laboratory – d.m.n. L.V.Didenko.

After end of a laboratory part of research have started experiment on animals.

For carrying out of an experimental part we had been developed experimental model of critical defect in a calvaria of a rat. The experimental model of defect used by us, according to foreign researchers, is as much as possible informative and approached to natural conditions of functioning of an osteoinductive biomaterial in an organism of the recipient. Modelling of process of a restore of defect gives the chance to investigate density, volume and quality formed regenerata and to estimate osteoinductive characteristics in digital value. As

Influence rekombinantnogo human morphogenetic fiber of a bone rhBMP-2 in structure nedemineralizovannogo an osteal matrix on process of an osteogenesis both in a surgical intervention zone, and in a segment as a whole was investigated.

In experiment 54 puberal males of white rats of breed Vistar with mass of a body 400-500 g at the age of 4-5 months are used. Animals have been parted on 3 groups: I-я - control, II th – group of comparison and III th – experimental (on 18 rats in everyone). At animals of II th (experimental) group defect filled pure nedemineralizovannym with an osteal matrix

"osteodent", and at III (experimental) groups defect filled nedemineralizovannym with the osteal matrix "osteodent" containing rhBMP-2. In control group defect healed under a clot. Animals were deduced from experiment in following terms: 1, 2, 3 months, on 6 rats from each group.

During the postoperative period an animal made antiseptic processing of a wound by a solution hlorgeksidina 0,05 % for the first, third, fifth and seventh days. At animal all groups seams took out for the seventh days after operation. At survey noted signs of the postoperative inflammatory phenomena and spent an estimation of a current of a wound process for the first, third, fifth and seventh days after operation and brought result in tables. The clinical picture has been estimated on a set of clinical signs which estimated on system "Yes" and "is not present", that corresponded to points 1 and 0, and expressed in a percentage parity.

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Table 8

Frequency of occurrence of clinical signs in the postoperative period

Clinical sign (symptom)

Control group Comparison group Experimental group
Days Days Days
1 3 5 7 1 3 5 7 1 3 5 7
% Signs % Signs % Signs
Fervescence 66,7 33,3 11,1 5,6 94,4 77,8 38,9 16,7 94,4 83.3 33,3 11,1
Edema of soft tissues 72,2 55,6 33,3 11,1 83,3 77,8 44,4 0 72,2 83,3 38,9 0
Augmentation regionarnyh lymphonoduses 0 0 0 0 5,6 5,6 5,6 5,6 5,6 5,6 5,6 5,6
Hyperemia presence 11,1 5,6 5,6 0 27,8 66,7 27,8 11,1 27,8 61,1 27,8 5,6
Incompetence of seams 0 0 0 0 0 0 0 11,1 0 0 0 5,6
Hematoma presence 0 0 11,1 11,1 0 0 0 0 5,6 5,6 5,6 0
Wound pyesis 0 0 0 0 0 0 0 11,1 0 0 0 5,6

For the first days the difference of frequency of occurrence of clinical signs in experimental group and comparison group has made 0,8 % in favour of experimental group.

For the third days has made 3,2 % in favour of experimental group. For the fifth days has made 0,8 % in favour of experimental group. For the seventh days has made 3,2 % in favour of experimental group.

After the spent analysis of the received data of expression of clinical implications at laboratory animals after an intervention of statistically significant differences between group of comparison and experimental group it has not been taped. Hence, presence rhBMP-2 as a part of an osteal material does not influence degree and a current of inflammatory process.

Histological research of samples of the calvaria including area of a surgical intervention, spent immediately after a biopsy. A material placed in 10 % formalin solution on the phosphatic buffer at 72 o'clock then within 24 hours samples of a tissue washed out in flowing water. A decalcification spent in the electrolytic way in oversaturated solution EDTA. On termination of process samples of a tissue washed out in flowing water within 24 hours. The slices washed out and dried on a filter paper placed at 24 o'clock in 50 % an ethanol solution.

As a result of the carried out morphological research it is established, that implantation of a material with rhBMP-2 shows a difference with comparison group in the first 2 terms of observation (1 and 2 months) aside to higher efficiency of an osteogenesis in comparison with implantation of a material not containing rhBMP-2, and also the used model of critical defect of an osteal tissue is adequate and answers the demand to objects in view and problems (Bull-calves A.I., 2013).

Proceeding from the received data of a computer tomography, differences between control and skilled groups in favour of the last are taped. Between experimental group and group of comparison of statistically significant differences it was revealed not.

Hence, presence rekombinantnogo human morphogenetic fiber of a bone rhBMP-2 as a part of an osteoplastic material does not influence the area of the restored osteal tissue. That concerning differences between control and skilled groups it is possible to explain it to that the generated osteal defect d=8мм. In a calvaria of a rat is critical and has no possibility to be restored independently.

The researches carried out by us and estimation of osteoinductance domestic rekombinantnogo human morphogenetic fiber of a bone rhBMP-2 has shown high prospect of its clinical use together with various combinations of osteoplastic materials.

Adhesion of a cellular material on a surface of the osteoplastic material sated rekombinantnym with human morphogenetic fiber of a bone rhBMP-2 proves expediency of application for filling of osteal defects and use for reconstructive operations on gnathic bones in CHLO.

Data of morphological researches of studied segments allows to do a conclusion about high-grade structure osteal regenerata, formed in defect. Further it will allow is high-grade and optimum to use the given site in gnathic bones for various kinds of treatment, including dentalnoj implantations.

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Scientific source Doliner Michael Ellevich. Studying of osteoinductive activity of the osteoplastic materials containing rekombinantnyj morphogenetic fiber of a bone rhBMP-2, in experimental models. The dissertation on competition of a scientific degree of the candidate of medical sciences. Moscow - 2014. 2014

Other medical related information discussion:

  1. CHAPTER 4. DISCUSSION
  2. Chapter 4 Discussion of the received results
  3. DISCUSSION OF THE RECEIVED RESULTS
  4. DISCUSSION OF RESULTS OF OWN RESEARCH
  5. Chapter 7 DISCUSSION of RESULTS of RESEARCH
  6. CHAPTER 8. THE CONCLUSION (DISCUSSION OF THE RECEIVED RESULTS)
  7. CHAPTER №5 DISCUSSION
  8. 2. RESULTS AND THEIR DISCUSSION
  9. RESULTS OF RESEARCH AND THEIR DISCUSSION
  10. Chapter 5 Discussion of the received results
  11. Chapter 7 DISCUSSION of RESULTS of RESEARCH
  12. CHAPTER 4. THE CONCLUSION (DISCUSSION OF THE RECEIVED RESULTS)
  13. Chapter 7 DISCUSSION of RESULTS of RESEARCH
  14. CHAPTER 4. DISCUSSION of the RECEIVED RESULTS
  15. the Chapter VI. DISCUSSION of the RECEIVED DATA