RU2283122C1 - Method for treating imperative incontinence of urine - Google Patents

Abstract

FIELD: medicine, gynecology.

SUBSTANCE: one should introduce for a patient twice monthly 50 mln fetal stem mesenchymal human cells intravenously and 6x10⁹ fetal stem neutral cells intramuscularly. The innovation enables to obtain stable curative effect in short terms at minimum of contraindications at no side phenomena.

EFFECT: higher efficiency of therapy.

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Description

The present invention relates to therapy and may find application in the treatment of imperative urinary incontinence.

Urogenital disorders in menopause are a complex of vaginal and urinary symptoms, the development of which is a complication of atrophic and dystrophic processes in estrogen-dependent tissues and structures of the lower third of the genitourinary tract: bladder, urethra, vagina, ligamentous apparatus and pelvic floor muscles.

The basis of the therapy of imperative urinary incontinence is hormone replacement therapy in combination with one of the drugs with M-anticholinergic action, such as oxybutynin (novitropan), detruzitol (tolterodine), trospium chloride (spazmeks) (Russian Drugs Encyclopedia, 10th edition 2003, pp. 611, 265, 852).

The main disadvantage of such therapy is a large number of side effects, such as dry mouth, nausea, vomiting, constipation, flatulence, decreased motility of the stomach and intestines, palpitations, tachycardia, vasodilation, weakness, dizziness, drowsiness or insomnia, anxiety, hallucinations, decreased production lacrimal fluid, mydriasis, increased intraocular pressure, accommodation paralysis, visual impairment (amblyopia), decreased sweating, suppression of lactation, allergic reactions (rashes) and many contraindications.

A partial reduction in side effects of such therapy was achieved through the use of vaginal suppositories containing oxybutynin and estriol (RF Patent No. 2241442, 2004). The duration of therapy is at least 6 months.

However, this tool does not completely eliminate the above side effects. In addition, therapy using this remedy is lengthy.

The objective of the invention is a method of treating imperative urinary incontinence, which has no side effects and has a minimum of contraindications, which has a shorter duration.

The problem is solved by the method consisting in the fact that the patient is injected intravenously with 40-60 million human mesenchymal stem cells and subcutaneously or intramuscularly 5-7 × 10 ⁹ fetal neural progenitor cells.

Obtaining and culturing mesenchymal cells. The bone marrow from the fruits of the 2nd trimester of gestation (18-23 weeks) served as a source of material.

A heparinized bone marrow aspirate was diluted with a 1: 1 Hanks solution. Ficoll / Pak was fractionated on a gradient (1.073 g / ml). Cells from interphase were transferred to Hanks' solution, precipitated at 1000 rpm for 15 min and resuspended in complete medium (MEM without deoxyribonucleotides or ribonucleotides, 20% fetal calf serum, 100 u / ml penicillin, 100 μg / ml streptomycin, 2 mM L -glutamine). Cells were cultured at 37 ° C, 5% CO ₂ . After 24 hours, non-adherent cells were removed, and adherent cells were washed with PBS (2 times). Once every 3-4 days, the environment was partially changed. Upon reaching the monolayer, the cells were scattered by incubation in 0.05% trypsin at 37 ° C for 3 minutes. Spent 2-6 passages to obtain the required number of cells.

Obtaining and culturing neural cells.

The source of cellular material was the brain of human embryos of the 1st trimester of gestation (8-12 weeks) and the periventricular zone of the fetuses of the 2nd trimester of gestation (17-23 weeks).

Brain tissue was suspended with a disposable syringe in a chilled (+ 4 ° C) saline solution. The resulting cell suspension was centrifuged at 1000 rpm for 3 minutes and washed twice with F12 medium. The pellet was resuspended in serum-free DMEM \ F12 medium (1: 1) with the addition of basic fibroblast growth factor (hFGF-b) -20 ng / ml; epidermal growth factor (hEGF); neuronal cell survival factor (N-2 supplement) -1: 100; heparin - 8 μg / ml; penicillin / streptomycin - 20 ng / ml. The final cell concentration in the medium was 2 × 10 ⁶ cells / ml. The cell suspension was cultured at 37 ° C, 5% CO ₂ . A partial change of medium (1: 1) was performed 2 times a week, while cell aggregates were dissociated by pipetting. On the 7th – 9th day of cultivation, multiple neurospheres formed, the sizes of which increased during cultivation. Cell counting was carried out in the initial suspension and with subsequent dissociation of cell aggregates in the Goryaev chamber. Cell viability was determined by a trypan blue standard assay.

The following example illustrates the method of the invention.

Example 1. Patient S., 56 years old, postmenopause 7 years, complaints of frequent daytime (up to 12 times), frequent nightly (up to 5 times) urination, urinary incontinence during urination.

Diagnosis: imperative urination disorder. A comprehensive urodynamic study revealed a decrease in detrusor pressure to 5-20 cm of water. I received detrusitol 1 mg 2 times a day for 1.5 months, noted poor tolerance (dry mouth), spasmex 15 mg 2 times a day, and frequent nightly urination and dry mouth remained. The patient was prescribed suppositories containing oxybutynin and estriol. Dry mouth disappeared, but frequent urination persisted.

The decision was made to introduce stem cells. A patient twice with an interval of 1 month was intravenously injected with 50 million human mesenchymal stem cells and intramuscularly 6 × 10 ⁹ fetal neural progenitor cells. The treatment effect was noted already in the 2nd week after the 1st injection.

A persistent therapeutic effect was noted within 6 months of observation.

Claims (1)

A method of treating imperative urinary incontinence in women, which consists in the fact that the patient is injected twice, with an interval of 1 month, intravenously 50 million fetal stem mesenchymal cells of the person and intramuscularly 6 × 10 ⁹ fetal stem neural cells.