Histo-equivalent bioplastic material. RU patent 2513838.

FIELD: medicine.

SUBSTANCE: what is described is a histo-equivalent bioplastic material comprising a carrier in the form of a matrix that is presented by a native form of hyaluronic acid. The histo-equivalent bioplastic material is prepared by mixing 1.5% hyaluronic acid and 5% peptide complex in the following proportions: 80-90 ml and 10-20 ml, respectively to form a viscous elastic gel that is placed on a mould carrier and exposed to ultraviolet photopolymerisation in a laminar flow hood for 5-7 hours, and then transferred onto a perforation apparatus with the finished material being perforated and notched.

EFFECT: more effective wound healing.

1 tbl, 11 dwg, 1 ex

The invention relates to medicine, namely to combustiology, surgery, cosmetology and can be used as bioplastic material for the substitution of the defects integumentary tissues (protection from evaporation and infection), and stimulation of regeneration.

The development and study of new biodegradable plastic and biocompatible materials for effective organ-specific regeneration with high functional and aesthetic result is the actual problem in modern regenerative medicine and Transplantology. Obviously, new materials should meet the compliance requirements of the particular morphology of the recipient tissues and promote their functional recovery (Bioartificial organs, 1999; Biocompatibility, 1999; Sudesh et al., 2000; 2004; Biopolymers for Medicinal and Pharmaceutical Applications, 2005).

Currently actively developing the newest direction of medical bioengineering to create tissue-engineering structures and bioisosteric bodies on the basis of biomaterials with new functional properties, the so-called gesticulant-bioplastic materials (BBB) (Shumakov, 1995; Shumakov et al., 2003; Stilman, 2006). A key feature of these materials is their ability to biodegradation natural metabolic pathways in the body with the inclusion of intermediate and final products in biochemical cycles without their system and local accumulation, such as lactic and glycolic acid included in the Krebs cycle. However, such products must not be toxic, and their concentration in the blood line must not exceed the maximum permissible level (Volova YEAR, 2003).

Physiological metabolizing biomaterials that make up a frame basis tissue-engineered constructs, determines the balance of reparative processes without expressed events inflammatory reactions and prevents the phenomenon of immune rejection, while avoiding the body's response to a foreign body (shyshatskyy that is, 2011).

The development of new gesticulant-bioplastic materials (BM) is based on the study of the kinetics of birthrate and dynamics of its mechanical properties, as well as on assessment of the impact and nature of the regenerative process. The nature and degree of severity of this impact are determined by the totality of physical-chemical properties of the actual material and intensity of the response physiological and biochemical reactions of the recipient organism.

Therefore, development of new biodegradable materials with the maximum degree of biochemical complementarity is based on creating a matrix consisting of macromolecular complexes available for private enzyme systems of the body and other lytic agents.

Therefore perfect biodegradable material must meet the following requirements.

1. Macromolecular design with the programmed period biodegradation natural metabolic pathways, which is not the object of immuno-inflammatory reactions.

2. The inclusion of intermediate and/or final products of biomateriali material in regeneration mechanisms on the phase of the signal chemotaxis protective cells.

3. Maximum compliance with the time period biodegradation of material and duration of the reparative process.

Thus, from the perspective of optimal immune-biochemical of compliance the implementation of the above requirements for the development of new biodegradable materials ensure optimum morphological and functional result organ-specific histogenesis.

Early research on the development of biodegradable materials were focused on natural polymers (collagen, cellulose and others), subsequently - on products of chemical synthesis. Examples of such biodegradable polymers are polianytzia, polyesters, high, poly (methylmethacrylate), polyurethanes. There are several key factors that allow you to control the dissolution of the material: the hydrophilic/hydrophobic, amorphous/crystallinity, molecular weight, presence of heteroatoms (for example, in addition to carbon) (Khlusov I.A., 2007).

Naturally, the most promising are the materials, the splitting of which formed a natural monomers. For example, polylactide, polyglycolides, polyoxyalkylene and their copolymers split accordingly to lactic, glycolic, hydroxybutyric acid, of which the Krebs cycle form water and carbon dioxide, deriving from the body naturally.

The prototype of the present invention is nanostructured bioplastic material (RF Patent№2425694 publ. 10.08.11 year), which includes native form of hyaluronic acid, and based on nanostructured matrix representing nanostructured hyaluronic acid, obtained by photochemical knitting, with a honeycomb structure in the range from 50 to 100 nm.

Such structural organization of macromolecules of hyaluronic acid and collagen gives the biomaterial elasticity, increased adhesion, drainage quality, transparency.

However thus obtained macromolecular structure bioplastic material in the clinical setting is not effective enough.

1. The structure of this material is monophasic, resulting in conditions of wound process it forms a homogeneous floor, becoming, thus, in a dry scab (Rakhmatullin P.P. Bioplastic material on the basis of hyaluronic acid: biophysical aspects of pharmacological properties. // Pharmacy. - 2011. - №4. - P.37-39). Feedback from clinicians, homogeneous dry biological scab requires daily dressings with mandatory hydrate formation, which ultimately leads to the prolongation of terms of healing and cicatricial changes with restriction of functions, for example in the joints.

2. Complex nanostructure organization of biomaterial significantly complicates the process of biomateriali in the wound, i.e. in process of healing it is not absorbed and becomes the reason for the accession of secondary infection, and as a consequence, the complicated course of wound process. Accordingly requires removal of material from the wound with dressings, and as the dry scab firmly spaevaet with subject tissues, this procedure is traumatic to the wound and painful for the patient.

3. Monophasic nanostructured organization of biomaterial is not effectively drainage of purulent and leads to the accumulation of fluid beneath the biomaterial, which is necessary for dressings additional material to punch a scalpel and shape it drainage window (Rakhmatullin P.P., burlutskaya I., Adelshin LR, Burtseva TI The effectiveness of a new method of restoration of the skin defect in a patient with congenital epidermolysis bullosa: clinical observation. // Pediatrics. - 2011. Volume 10, Number 2, - S-192). Such manipulations "disturb" the wound and painful tolerated by the patients, especially children.

Thus, the nanostructuring bioplastic material causes the formation of optimal bioengineering properties (adhesion, transparency), but does not provide favorable wound healing and may cause complications.

Effect : increased efficiency of wound healing.

The problem is solved by the fact that in gesticulant-bioplastic material, including a base in the form of a matrix, as the material which is used native form of hyaluronic acid, according to the invention gesticulant-bioplastic material contains 1.5% solution of hyaluronic acid 5% solution peptide complex, mixed before the formation of viscous elastic gel placed the basis and subjected to ultraviolet photopolymerization in laminar cabinets for 5-7 hours at the following quantitative ratio, ml:

- 1.5% solution of hyaluronic acid - 80-90;

- 5% solution of peptide complex - 10-20,

while the team has perforation and notches.

Figure 1 presents microelement gesticulant-bioplastic material, figure 2 - distribution of forces of adhesion to the surface of gesticulant-bioplastic material, figure 3 - relief similarity of biomaterial with skin human figure, figure 4 - the scheme of two-phase structure of biomaterial, located in the wounds, figure 5 - atomic spectrometry image biomaterial after cell cultivation, figure 6 - location scheme of notches and holes gesticulant-bioplastic material on Fig.7 - view trophic ulcers of the left tibia patient diabetes, Fig and 9 - stages bioplastics ulcers left Shin patient diabetes, figure 10 and 11 - the stages of healing of ulcers left Shin patient diabetes.

The structure of the peptide complexes present in the hydrocolloid of hyaluronic acid, is presented in the table, where it is seen that the peptide complexes have different amino acid composition with molecular weight ranging 244-459 Yes. In detected peptides prevail aliphatic (leucine, isoleucine, alanine, glycine) and polar uncharged amino acid residues: threonine, Proline, histidine, serine, and polar charged amino acid residues: arginine, glutamine, asparagine, lysine, arginine. In addition, there are dimer isoleucine and polymer tripeptides, including peptides containing aromatic amino acid residues (tryptophan) and polar uncharged amino acid residues.

The analyzed parameters

Chem. formula

Delta mass in Nanomolecular mode

Mass in Rel. units

GlyTrpIle

C19H26N4O4

374.19541

IleAspIle

C16H29N3O6

359.20564

PheArgPro

C20H30N6O4

418.23285

GlnHisHis

C17H24N8O5

420.18697

AlaTrpLys

C20H29N5O4

403.22195

ProHisTyr

C20H25N5O5

415.18557

ThrTrpTrp

C26H29N5O5

491.21687

LysPheThr

C19H3ON4O5

394.22162

LysArgMet

C17H35N7O4S

433.24712

PheCysMet

C17H25N3O4S2

399.12865

C12H24N2O3

244.17869

AspLysLys

C16H31N5O6

389.22743

C16H19N3O3

301.14264

-At 18.38

C9H16N2O6

248.10084

Desmosine

C24H4ON5O8

526.28769

It is important that in peptide fractions present desmosine (an amino acid, a derivative of lysine). Due to its complex structure, which has four amino acid group, one molecule of desmosine can be in four peptide chain. Thus it is possible to form two-phase structure of gesticulant-bioplastic material.

In addition, two-phase structure of the new biomaterial due to the difference of forces tension gives the biomaterial unique macroreliefs figure. Metropolitanate gesticulant-bioplastic material biomaterial has a unique relief and looks very similar to dermatoglyphics of human skin.

Due to the different polarities of amino acids is the effect of the surface energy of tension, which affects the formation of the unique mikroelementi.

Atomic spectrometry images ultrastructure surface preparation presents globular formations similar morphology with a unique relief.

It is known that the consolidation of somatic cells is likely to happen on the surface of the material, possessing high surface energy (on a hydrophilic surface), at the same time on basic cellular processes (growth, differentiation, migration) largely influenced by the geometric and dimensional features of the relief of the substrate [Hertz N. Uber die Beruhrung Fester Elastischer Korper (On the contact of elastic solids) // J. Reine Angew. - 2011. - №92. - S. 156 171].

Assessing hydrophilic/hydrophobic properties of the proposed biomaterial method of fixation of the contact angle of the water, the value of which amounted to 83°, calculated on this basis the work of adhesion, which considering the roughness coefficient was equal 99,88 mn/m, which characterizes the surface of the new material as a moderately wet.

Additionally performed visualization of the surface of biopolymer in the registration regime forces of adhesion helped to narrow down the field with high adhesion.

The surface energy of the forces of adhesion is a fundamental characteristic of bioplastic materials for assessment of effective migration of cells on the surface during regeneration. It is established that the presence of local areas with different adhesion ensure taxis cells and uniform their distribution all over the surface (Hallab N.J. Bundy K.J. O'connor, K. et al. Evaluation of metallic and polymeric biomaterial surface energy and surface roughness characteristics for directed cell adhesion // Tissue engineering. - 2001. - V.7. - №1. - P. 55-71).

Estimating on the edge angle of wetting thermodynamic the work of the water on the surface of gesticulant-bioplastic material recorded values of the adhesion forces characterize his two-phase biomaterial with hydrophilic/hydrophobic properties. This is explained by the presence of different substances (amino peptide complex, spatially distributed in the structure of hydrocolloid hyaluronic acid that acts as a basis of the matrix material.

Confirmation of this was a direct visualization of the biomaterial surface mode adhesive-contact atomic force microscopy. The results indicate significant in relation to the visualized area on the biomaterial surface, demonstrating adhesive properties.

In turn, the cultivation of mesenchymal stromal stem cells using gesticulant-bioplastic material as a substrate and subsequent atomic-force microscopy surface has allowed to detect the presence of the cells of the oblong form, of a width 3,7 mm. Further more detailed research revealed that the cell surface is interwoven fibrillar fibers. The similar nature of their arrangement is a sign of that in cultured mesenchymal stem cell migration processes in which they actively interact with the surface of the substrate, penetrating into the subject matrix.

Thus, the presence of peptide complex provides the organization of two-phase structure of the biomaterial which is the basis for the formation of full biological scab in the wound (with the outer surface of the plate, inside - hydrocolloid). Gesticulant-bioplastic material has through the Microperforation for effective drainage of the wound.

Maximum morphological similarity in the structure and appearance of biomaterial with integumentary tissues (skin) gives grounds for introduction in the title of the invention of the term "gesticulant-bioplastic material".

Gesticulant-bioplastic material obtained as follows.

As raw material use hyaluronic acid native form, with fibrous nanostructured structure capable of forming an elastic plate and peptide complex. The native form of hyaluronic acid creates optimal conditions for the migration and proliferation of cells, the source of which are the pieces of viable tissue, placed in the region, however (a kind of cell to cell) gesticulant-bioplastic material.

Prepare a 1.5% solution of hyaluronic acid 5% solution of peptide complex at room temperature. Then they are mixed prior to the formation of viscous elastic gel.

The viscous elastic gel is placed on the form-based and engineered safety Cabinet subjected to ultraviolet photopolymerization.

To do this, in the BSC is created certain microclimate (temperature is 0 to 3 degrees centigrade, humidity - 50-55%, with ventilation air flow rate - 0,50 m/s). Then the mixture is subjected to ultraviolet irradiation for 5-7 hours. The ready material is transferred to the machine for punching and packaging.

In the end, in the forms smooth film whitish color, as similar as possible to the dermatoglyphics picture of human skin.

The plate by cutting given oval shape (large diameter 15 cm, the small diameter - 11 cm)square, oval plate corresponds to the average area of a palm of an adult. In medicine it is believed that the palm of an adult person is about 5% of the body surface, for example, thus, is estimated area of burns.

Then on the plate with the gun applied the round hole, and then mechanically using a special knife. Evenly distributed incisions required for drainage of the wound exudate and more dense connection of biomaterial and the underlying tissue. Round holes with a diameter of 3 mm serve bore holes for epithelial tissues. These fabrics are own tissues of the patient that excising from the edges of the wounds in her surgical treatment. They are used as sources of cells for effective epithelization (healing) of the wound.

As a result of clinical use in patients with defects integumentary tissues found that gesticulant-bioplastic material forms a full biological scab and accelerates wound healing.

It is important to note that gesticulant-bioplastic material was effective in patients who traditional therapy did not help.

Clinical example.

Sick A., 59 years old, lives in Kuvandyk, entered the 4 x/NPH "OKB on Art. Orenburg, JSC "Russian Railways" on 16 December 2011 with a diagnosis of Giant circular purulent-necrotic trophic ulcer of the left tibia. Diabetes type 2 diabetes, severe. Allergic dermatitis lower extremities. Anemia.

Sick for 1 year, how did the plague after suffering necrotic form faces. The size of the defect circular swath width of 8 to 10 cm from n/a 3 to p/3 shank with elements of fascia and ligaments in bottom of the wound. All previously used methods of treatment, including bandages with antiseptics, ointments and a variety of wound coverings effect has not had and extremely painful tolerated by the patient.

Upon receipt of the executed debridement, hewed from necrotic lesions, with getting round pieces of viable tissue of the skin, size 0.5 to 1 mm Then on the prepared wounds impose gesticulant-bioplastic material, having in the structure of Microperforation 3 mm in diameter and notches. After the biomaterial will stick to the wound and become elastic plate in his perforation fit retrieved from saline pieces viable tissues of the patient. Once all of Microperforation will be filled with pieces of cloth, laid sterile napkin and dressing.

Spent a long course of a complex of conservative treatment with correction of comorbidity. We observed formation of two-phase structure of biomaterial, located in the wound: an external plate - biological scab, and internal - hydrocolloid.

After cleansing the wound was formed flat granulating circular defect with edge epithelialization.

When using gesticulant-bioplastic material was established that it has the most powerful stimulating effect on fibroblasts, activating receptors CD44 on the synthesis already own hyaluronic acid, type III collagen and elastin, which prevents the formation of hypertrophic scars.

In the result of the treatment offered by the biomaterial noted that in 2-3 hours after applying all patients reported no pain. Subsequently, it was noted the absence of the phenomenon of the contracture changes wound area and full restoration of the skin. It should be noted that the use of this biomaterial has allowed to avoid inflammatory reaction (abscesses) and formation of hypertrophic scars in the rehabilitation period. Not recorded a single case of allergic and/or inflammatory response in patients.

Thus, in comparison with the prototype conducted investigations show that gesticulant-bioplastic material is able to form a proper biological scab and stimulate healing of wounds with good aesthetic result without scarring, and has high biocompatibility with integumentary tissues the human body.

The novelty is developed gesticulant-bioplastic material is original line-peptide complex, with different amino acids. The availability of this peptide complex provides the organization of two-phase structure of the biomaterial which is the basis for the formation of full biological scab in the wound (with the outer surface of the plate, inside - hydrocolloid). Gesticulant-bioplastic material has through the Microperforation for effective drainage of the wound.

The distinctive feature of this gesticulant-bioplastic material is its ability to form a two-phase wound floor and form so full biological scab, effective drainage of the wound, which ultimately ensures optimal the regeneration of the defect cover tissues without scarring and deformation. In addition, the biomaterial has a special boarding holes for the patient's own tissue, which are formed sprout area for wound healing.

Gesticulant-bioplastic material, including a base in the form of a matrix, as the material which is used native form of hyaluronic acid, wherein specified bioplastic material obtained by mixing of 1.5% solution of hyaluronic acid and 5% solution of peptide complex in the following quantitative ratio: 80 to 90 ml and 10 to 20 ml, accordingly, before the formation of viscous elastic gel, which is placed on the form-based and subjected to ultraviolet photopolymerization in laminar cabinets for 5-7 hours, with the subsequent transfer to the machine for punching, while the ready material has perforation and notches.