RU2236240C1 - Method for treating diabetes mellitus - Google Patents

Abstract

FIELD: medicine, endocrinology.

SUBSTANCE: one should implant cells of human benign insulinoma into pancreatic caudal tissue in diabetes mellitus-suffering patient. Before implantation one should conduct standardization of cell culture by the debit of synthesized insulin. The method provides decreased dosage of applied exogenous insulin in case of achieving normal values of glycemia in patients.

EFFECT: higher efficiency of therapy.

2 ex

Description

The invention relates to the field of surgery and therapy and can be used in the treatment of diabetes mellitus.

A known method of treating diabetes mellitus, comprising transplanting mammalian pancreatic β-cells (newborn piglets, rabbits, etc.) into a capsule formed by preliminary subcutaneous injection of a polyacrylamide gel, the cells are cultured using standard media and solutions (see IPC A 61 K 35/39, patent of the Russian Federation No. 2165263, publ. 04/20/2001).

A significant disadvantage of this method is the low effectiveness of the treatment, due to the need to continue treatment with oral antidiabetic drugs.

Closest to the claimed method for the treatment of diabetes mellitus is a method comprising implanting a cell culture of a benign human insuloma into the rectus abdominis muscle in the form of a suspension in saline solution, while the number of implanted cells is determined by the ratio N = Q: q, where

N is the number of implantable cells, thousand;

Q is the patient's daily need for insulin, units

Q is the daily productivity of insulin of 1 thousand cells, and when the cells are cultivated, their number and insulin content in the culture medium are measured at predetermined time intervals and the level of these values determines the insulin productivity of a given cell culture, and the number of culture cells is determined by this value, necessary for the preparation of the implant (see IPC A 61 K 35/39, patent of the Russian Federation No. 2004247, publ. 15.12.93) - the closest analogue.

Significant disadvantages of the known method for the treatment of diabetes are:

- the short duration of the production of insulin by the donor cells in the recipient organism due to the effect of rejection of heterogeneous cells, as a result of which significant immunosuppressive therapy and repetition of the transplantation operation are necessary;

- a decrease in insulin synthesis by graft cells due to the blast transformation of donor cells, leading to the introduction of exogenous insulin;

- the complexity of accurate control of insulin performance by a specific fraction of cell culture.

The technical result of the proposed method is to increase the effectiveness of treatment by eliminating repeated operations and conducting immunosuppressive therapy.

The essence of the technical solution lies in the fact that in a method for the treatment of diabetes mellitus, which involves implantation of benign human insulinoma cells, its standardization according to the debit of synthesized insulin according to the invention, implantation of benign human insulinoma cells is carried out in pancreatic tissue in its caudal part, while insulinoma tissue is before transplantation subjected to histological examination.

It is known that endocrine cells that produce insulin are located in the tail of the pancreas. The implantation of benign human insulinoma cells into the pancreatic tissue in its caudal part will exclude repeated transplantation operations by increasing the time the insulin donor cells produce into the recipient organism, eliminating heterogeneous cell rejection, and increasing insulin synthesis by transplant cells by eliminating cell blast transformation, which together increases the effectiveness of the healing process.

The introduction of a histological examination procedure for insulinoma tissue eliminates the possibility of transplantation errors of tumor cells of a different morphological structure and ensures the immune resistance of transplanted cells.

The method is as follows. In a patient with a human insulinoma, tumor enucleation is performed. The resulting tumor tissue is examined for its morphological correspondence to insulinoma tissue. After morphological confirmation of the presence of insulinoma tissue, the connective tissue capsule is removed and the insulinoma tissue is homogenized mechanically under sterile conditions. The homogenate obtained under sterile conditions is incubated in a thermostat at T = 37 ° C for two hours in a lactalbumin hydrolyzate with the addition of 5% fetal serum. After incubation, the homogenate is cooled to T = 4 ° C, placed in Earl's solution with the addition of antibiotics for 1.5-2.0 hours, washed until the supernatant is completely transparent and the pH is normalized.

Using a centrifugal method, a pure suspension of insulinoma cells is isolated from the homogenate prepared in this way, which is placed in a protective nutrient medium RPM - 1640 with the addition of 20% fetal serum and preserved at T = 4 ° C and a hydrostatic pressure of 400-500 atm.

Before implantation, the suspension of insulinoma cells is purified from the preservation medium by repeated washing in saline and placed in sterile physiological solution with the addition of a 5% glucose solution, the temperature is increased to 20-37 ° C, and the hydrostatic pressure is reduced to 760 mm Hg. Art.

The resulting insulinoma cell culture is standardized for 10 synthesized insulin debit. per day and implanted in the tissue of the tail of the pancreas of a patient with diabetes.

Implantation is carried out by the pneumoperitoneal method through the abdominal wall using fiber optics and an endoscopic instrument.

Example 1. Patient L. is observed for 8 years with a diagnosis of insulinoma, benign hormone-active. May 12, 1999, tumor enucleation was performed operatively. The resulting tumor tissue morphologically corresponds to benign insulinoma tissue. Tumor tissue was processed according to the proposed method. The resulting suspension of insulinoma cells was tested for hormonal activity before direct transplantation to the recipient 04/06/2000

Patient K., 33 years old. Type I diabetes mellitus, diabetic angiopathy of the vessels of the lower extremities. The daily requirement for exogenous insulin is 41 units, the fasting glycemia level is 15-17 mol / l. 04/06/2000, allotransplantation of insulinoma cells into the caudal part of the pancreas was performed using a puncture method in an amount of 5 ml of suspension and 40 units of hormonal activity. synthesized insulin using fiber optics and an endoscopic instrument with pneumoperitoneal access through the anterior abdominal wall. On the 4th day after transplantation, fasting glycemia of 3.5-3.8 mmol / l against the background of the basic administration of exogenous insulin at a dose of 20 units. and bolus administration of humalogue, ultra-short-acting insulin. On the 12th day, the basic administration of exogenous insulin, fasting glycemia of 4.8 mmol / l, the bolus administration of the humalogue was continued. On the 16th day, the patient was transferred to outpatient monitoring amid an improvement in general condition. Fasting glycemia of 6.7 mmol / l, continued administration of ultrashort insulin is recommended. The patient was observed for 12 months. Fasting glycemia did not exceed the upper limit of normal. The patient uses the bolus method of administering the humalogue, with dietary disorders not more than 2-3 times a month.

Example 2. Patient M., 47 years old, diagnosis: type I diabetes mellitus, insulin-dependent, diabetic angiopathy, nephropathy, arterial hypertension. The daily requirement for exogenous insulin is 60 units. per day. Fasting glycemia 14-16 mmol / l. The state of subcompensation, blood pressure 180/90 mm RT. Art. 04/14/2000, under general anesthesia, an allotransplantation of a culture of insulinoma cells into the tail of the pancreas was performed using a puncture method in an amount of 7 ml of a suspension with hormonal activity of 60 units. exogenous insulin by pneumoperitoneal access through the anterior abdominal wall using fiber optics and the use of endoscopic instruments from OLIMPUS. On the 3rd day, fasting glycemia sharply decreased to 2.7 mmol / L. The patient had a hypoglycemic state. The basal administration of exogenous insulin was discontinued due to the bolus administration of ultrashort insulin on the 12th day after transplantation. Against the background of an improvement in general condition, a decrease in the level of protenuria, a decrease in blood pressure to 140/80 mm RT. Art., the patient was transferred to outpatient observation and for 12 months he was observed by an endocrinologist. Fasting glycemia within the upper normal range. The patient was not given a bolus of ultrashort insulin.

The subjective and objective symptoms of the above patients, the data of blood tests, urine tests, and the general condition allow us to conclude that the proposed treatment method is highly effective up to the complete cure of patients suffering from exogenous insulin deficiency.

Unlike the known methods, the claimed technical solution also eliminates the possibility of malignancy of transplant cells and other side effects.

Claims (1)

A method for the treatment of diabetes mellitus, comprising implantation of benign human insulinoma cells, its standardization according to the debit of synthesized insulin, characterized in that the implantation of insulinoma cells is carried out in the tail of the pancreatic tissue.