Method of promoting the healing of wounds

 

(57) Abstract:

The invention relates to medicine and can be used to stimulate wound healing. As drug use supernatant from regenerating spleen, taken at the peak of reparative processes after resection. The method allows to reduce the time of healing without the formation of unsightly scars. table 2.

The invention relates to the field of experimental medicine, and it can be used to accelerate healing of wounds.

Known methods of stimulation of reparative processes by local application of drugs derived from the spleen of various animals: homogenate of xenoculture (Aliev, M. A. Using homogenate of xenoculture in the treatment of purulent wounds and ulcers.: Abstract. dis. Kida. the honey. Sciences. - Makhachkala. - 1994. - S. 16), slices and xenoplanet spleen (Stolyarov E. A., A. Sonis, Local application of drugs porcine spleen in the treatment of purulent wounds // efferent Methods and quantum therapy in clinical practice: abstracts. reports of the scientific-practical conference, may 25-26, 1995 - Izhevsk, 1995. - S. 178) xenoplanet and cryopreservant (Andrukhovich F. F., Sitnikov C. A., Tsypin A. B. and other Treatment is Z. reports of the scientific-practical conference 25 - 26 may 1995 - Izhevsk. - 1995. - S. 250 - 252).

The above methods have significant disadvantages, namely, that when using drugs spleen containing xenocentric, this may cause unwanted immune reactions and primarily sensitization of the body that prevent re-use. In addition, xenophobia for the most part are factors that do not have specificity in relation to the regulation of reparative processes, so the functionality of the impact of donor spleen on reparative processes underutilized.

The essence of the invention lies in the fact that local use of the supernatant obtained from regenerating the spleen of an animal donor, taken at the peak of reparative processes, that is, 10 days after resection.

Fence spleen of the animal donor in a regeneration phase, provided by the invention, produced on the basis of the functional characteristics of the spleen to influence the regeneration of other organs, namely 10 days after resection, it notes the maximum lymphoproliferative activity and secretion of specific honey is a convenient form of universal introduction (local, parenteral and others) among different preparations obtained from the spleen. He is able to keep their properties for a long period of frozen at -20oC. in Addition, the technique of obtaining it much easier than other similar drugs, such as xenoplanet. During experiments on small laboratory animals were used two types of models: linear and dolnoslaska planar wounds. In the treatment of wounds was introduced supernatant standardized to protein (mg/ml) by the method of Lowry. The most pronounced clinical effect was obtained when treating a linear wound in mice using a total dose of 1.5 mg/mouse weighing 20 g, planar wound - 3.2 mg/mouse weight of 20, increasing the dose did not alter the clinical effect, decrease the dose to achieve the same clinical effect failed, that is, these doses the local impact was optimal.

The method is as follows. The animal donor perform resection of the spleen, on the 10th day take regenerating spleen, homogenize it in 0.9% sodium chloride solution on the basis of 6.5 ml of a solution of 1 g of tissue, centrifuged at 3000 rpm at 4 to 6oC for 20 min, the supernatant collected and during the year, standardizing on protein and is used topically to treat wounds.

Example 1. Mouse-hybrid (IAS + C57BL/6)F1weight 20 g under ether anesthesia after removal of wool in the back right of the spine and parallel to it was applied linear cutaneous wound length of 1.5 cm on the depth of the skin wound sutured discrete silk suture. 24 hours after injury, the wound was cut away all the supernatant obtained from regenerating the spleen of an animal donor, taken 10 days after resection of the spleen and standardized for protein by the method of Lowry dilution with 0.9% sodium chloride solution to a concentration of 0.75 mg/ml, 4 Ukolov near wounds on both sides of the seam 0.1 ml (0.4 ml per mouse - 0.3 mg of protein by Lowry) for 5 days.

Example 2. Mouse-hybrid (IAS + C57BL/6)F1weight 20 g under ether anesthesia after removal of wool in the back struck paleologou planar wound area of 100 mm2. 24 hours after injury, the wound was cut away all the supernatant obtained from regenerating spleen and standardized for protein by the method of Lowry dilution with 0.9% sodium chloride solution to a concentration of 1.6 mg/ml, 4 Ukolov near wounds 0.1 ml (0.4 ml per mouse - 0.64 mg protein for SUB>1with linear wounds of soft tissues and 25 - planar. In the experimental group with a linear model of the wound for 5 days was introduced supernatant regenerating spleen in a total dose of 1.5 mg/mouse weight of 20 g in the experimental group with the model plane wounds supernatant regenerating spleen in a total dose of 3.2 mg/mouse weight of 20 g in the control groups was 0.9% sodium chloride solution of 4 Ukolov near wounds 0.1 ml to 0.4 ml per mouse for 5 days. The dynamics of wound healing was observed within 14 days, controlled the healing of all wounds visually, planar RAS - planimetric method. At 7 and 14 days was resellable area of the skin at the site of a former injury for histological and tenzimetricheskikh (linear RAS) studies.

Planimetric studies have shown that the supernatant regenerating the spleen has a positive effect on the healing of planar Russian Academy of Sciences (PL. 1). In the experimental group the area of wound (S) 3, 5, 7, 10 days was significantly lower (p<0.05) and the percentage reduction in the area of RAS relative to the source (S0and on the previous day ( S3-5,5-7,7-10) higher during the whole observation period.

In the test method for laboratory belly of the experimental groups revealed typical changes typical healing "clean" wounds. In the group with drug use regenerating spleen observed a larger number of cells (macrophages and lymphocytes) in the area of the scar at the level of the muscle plate and in the underlying tissue. The effect is, we believe, stems from the fact that under the influence of factors regenerating the spleen is a fast and massive transport of macrophages, lymphocytes of the recipient in the injury area, and cytokines produced by these cells activate fibroblasts in the wound, thereby regulating the process of collagenopathy and healing. In addition, factors regenerating spleen can have a direct activating effect on fibroblasts.

Based on the fact that one of the stages of wound healing is the formation of scar tissue, examined its qualitative characteristics. Visually noted that in the experimental group of animals with a model of linear wounds in contrast to the control, healing occurred with the formation of a gentle, barely noticeable scars, having greater strength, which is confirmed by tenzometricheskie research samples regenerating skin areas (PL. 2). To 7 days in the control group discontinuous voltage - 0,130,008 MPa, in the basis of the strength of the scar in the main group. To 14 days in the control group burst voltage -0,320,008 MPa, in the main - 0,350,019 MPa (p>0,05). From the above it becomes obvious that the scar reaches strength in the main group in a shorter time than in the control.

The proposed method of promoting wound healing of soft tissues has the following advantages:

1. Accelerate healing of wounds due to the activation of reparative processes.

2. Is better wound healing with formation of a strong wealthy scars.

3. Wound healing is a cosmetic with the formation of a gentle, barely noticeable scars.

4. The proposed method can be used for the design of surgical intervention on any soft tissues.

Method of promoting wound healing by local application of the drug from the spleen of an animal donor, characterized in that as drug use supernatant obtained from regenerating spleen, taken at the peak of reparative processes after resection.

 

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FIELD: medicine, surgery, transplantology.

SUBSTANCE: embryonic spleen should be sampled, washed in nutritive medium № 199 to be placed into fresh medium № 199 to obtain homogenate in teflon homogenizer followed by centrifuging; then one should isolate the upper, medium and inferior layers, suck off medium layer and the upper part of inferior layer; the cell mixture obtained should be diluted in nutritive medium № 199 to be then introduced by injections into mesentery of small intestine or rectus muscle of abdomen. The present innovation favors the activation of immune system in patients undergone splenectomic operation and in those in case of surgical immunodefficient state due to high functional and regenerating activity of transferred embryonic splenic cells.

EFFECT: higher efficiency of prophylaxis.

6 dwg, 2 tbl

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