Background
Stress Urinary Incontinence (SUI) is a common disease of middle-aged and elderly women, the incidence rate in Europe and America is 10-23%, and the incidence rate in China is about 18.9%; the female stress urinary incontinence has the etiology that 1, after the age of 40, the female starts to decline the ovarian function, reduces the estrogen, and the epithelial cells start to shrink; 2. muscle damage during pregnancy and childbirth; 3. congenital dysplastic factors; the symptoms are that when the abdominal pressure is increased (cough, sneeze, laughing, heavy load and the like), urine cannot automatically overflow, which brings great trouble to the daily life of middle-aged and old women and leads to the obvious reduction of the quality of life.
Currently, treatments for female stress incontinence include physical exercise, medication and surgical treatment; physical exercise has certain curative effect when the symptom is slight, the treatment of the medicine is controversial due to large side effect and poor curative effect, and the operation treatment is the most effective method at present; the surgical treatment comprises three types of anterior vaginal wall repair, retropubic urethra bladder suspension and sling operation, and because of traumatic operation, improper operation can cause tissue damage, postoperative pain discomfort and urine retention and other side effects.
The injection therapy has the characteristics of simple and convenient operation, no wound, safety and repeated operation, and has the defects that the long-term curative effect is not better than that of the operation therapy, the injection filling material comprises polytetrafluoroethylene, collagen, silicone polymers, resin compounds, calcium hydroxyapatite, polyacrylamide hydrogel and the like, and compared with the operation therapy, the recurrence rate of the filler after the injection therapy is higher, although the short-term effect is better, the injection material is degraded along with the time, the long-term curative effect is obviously reduced, and the risk of complications such as aseptic abscess, granuloma foreign body formation, immunological rejection and the like exists.
Disclosure of Invention
The invention aims to provide an injection preparation for treating female stress urinary incontinence, wherein mesenchymal stem cells and a filling agent in the injection preparation in a short period of time play a filling role together to relieve female stress urinary incontinence symptoms, and simultaneously the mesenchymal stem cells stimulate local tissue repair and angiogenesis in a microenvironment through secreting growth factors and cytokines to enhance the contraction function of a urethral sphincter, so that the problem of female stress urinary incontinence is fundamentally solved.
The technical scheme for solving the technical problems is as follows: an injection preparation for treating female stress urinary incontinence is provided, which comprises mesenchymal stem cells and a filler; the volume ratio of the mesenchymal stem cells to the filler is 1: 1-1: 3.
the beneficial effects of adopting the above technical scheme are: in the injection preparation, the mesenchymal stem cells have wide sources, are easy to collect, have no ethical problem, are safe and have no tumorigenicity; because the low expression HLA class I antigen and the non-expression II antigen have the function of immunosuppression, the local tissue injection can reduce the levels of inflammatory factors TNF-alpha, IL-12, IFN-gamma and IL-5, enhance the production of anti-inflammatory factors IL-10, IL-4, TGF-beta and LIF, and has the function of local anti-inflammation; aiming at symptoms of urethral and bladder trigone submucosal vein thinning, blood supply reduction, mucosal epithelium degeneration, urethral and bladder superficial epithelial tissue tension reduction, urethral and peripheral pelvic floor muscle atrophy and the like of postmenopausal women caused by estrogen hypofunction, mesenchymal stem cells in the preparation promote nerve function recovery, promote angiogenesis and atrophic muscle repair and enhance the contraction force of urethral sphincter by secreting various growth factors and cytokines including VEGF, bFGF, PDGF, TGF and the like in a tissue microenvironment through paracrine effect, thereby fundamentally solving the problem of female stress urinary incontinence.
Meanwhile, the filler such as medical hyaluronate is safe, non-toxic and non-immunogenic, the hyaluronate naturally exists in a body and plays a role in moisturizing and lubricating, mesenchymal stem cells are cultured in vitro, and a proper amount of medical sodium hyaluronate is added, so that compared with a control group, the proliferation activity is enhanced, and the form and antigen expression are not abnormal; hyaluronic acid has a certain anti-inflammatory effect in addition to the moisturizing and lubricating effects.
In the injection preparation, the mesenchymal stem cells and the filling agent play a filling role together in a short period, so that the symptoms of female stress urinary incontinence are relieved, meanwhile, the mesenchymal stem cells stimulate local tissue repair and angiogenesis in a microenvironment through secreting growth factors and cytokines, so that the contraction function of the urethral sphincter is enhanced, and after the filling agent and the allogeneic mesenchymal stem cells are gradually decomposed and absorbed by an organism, the regenerated tissues play a role in relieving the female stress urinary incontinence for a long time, so that the problem of the female stress urinary incontinence is fundamentally solved; meanwhile, due to the immunosuppressive and anti-inflammatory effects of the mesenchymal stem cells, sterile inflammatory reaction and immune reaction caused by certain types of fillers are inhibited.
On the basis of the technical scheme, the invention can be further improved as follows:
further, the mesenchymal stem cells are derived from umbilical cord placenta, fat, dental pulp or endometrium of the parturient.
Further, the filler is hyaluronate, collagen, a resin compound, polytetrafluoroethylene, calcium hydroxyapatite, a silicone polymer or polyacrylamide hydrogel.
The injection method of the injection preparation for treating female stress urinary incontinence comprises the following steps:
injecting needle from the front wall mucosa of vagina 1.5cm away from the open end of vagina, pushing 0.5-1ml injection preparation into the gap between the front wall of vagina and the connective tissue of the back wall of the middle section of urethra respectively at the upper, lower, left and right 4 points with 0.5cm as radius and taking the needle-inserting point as the center, and retreating while pushing until a semi-surrounding is formed around the back wall of the middle section of urethra. The injection is injected at the part, so that the preparation can achieve the effect of lifting the middle section of the urethra in a short time and a long time, can relieve and eliminate the symptoms of stress urinary incontinence, and has small harm to patients and small side effect.
The beneficial effects of adopting the above technical scheme are: the injection method is simple and feasible, the injection preparation forms a semi-surrounding around the rear wall of the middle section of the urethra, the operation is safe and easy, no wound exists, no obvious side effect exists, the symptoms of female stress urinary incontinence can be relieved for a long time, and the normal life of middle-aged and old women can be recovered.
Further, the number of mesenchymal stem cells contained in a single injection dose is 1 × 107-5×107And (4) respectively.
Detailed Description
The invention is described below by way of example only, which is intended to illustrate the invention and is not intended to limit the scope of the invention.
Example 1:
an injection preparation for treating female stress urinary incontinence comprises mesenchymal stem cells and a filler; the volume ratio of the mesenchymal stem cells to the filler is 1: 1; the mesenchymal stem cells are derived from umbilical cord placenta donated by healthy lying-in women volunteers (24 hours after birth), are prepared, amplified and cultured on a cell preparation platform conforming to GMP standard until 3 rd to 5 th generations are frozen for standby use, and the quality inspection is qualified;
the filler is mainly sodium hyaluronate, the molecular weight of the filler is 150 ten thousand, and the filler is prepared into 1% concentration by using normal saline for later use;
taking umbilical cord mesenchymal stem cells 1 x 10 each time7And 3ml of the total volume, namely preparing 3ml of medical-grade sodium hyaluronate (with the molecular weight of 150 ten thousand) with the concentration of 1% from physiological saline, and mixing to obtain the medical-grade sodium hyaluronate.
When in use, the injection needle is injected from the mucosa of the anterior wall of the vagina at a position 1.5cm away from the open end of the vagina, the needle head enters the gap between the connective tissues of the anterior wall of the vagina and the posterior wall of the middle section of the urethra, then 0.5-1ml of injection preparation is respectively pushed into the upper, lower, left and right 4 points by taking the needle inlet point as the center and taking 0.5cm as the radius, and the injection preparation is pushed and retreated while being pushed until a semi-surrounding bag is formed around the posterior wall of the middle section of the urethra; wherein the number of mesenchymal stem cells in single injection dose is 1 × 107-5×107And (4) respectively.
The injection preparation can be repeatedly injected, 2-3 times are injected in each course of treatment, the interval is 1-2 months, and the treatment amount of the 2 nd to 3 rd times is determined according to the symptom relieving condition of a patient; the mesenchymal stem cells and the filling agent in the injection preparation of the invention play a filling role together in a short period, relieve the symptoms of female stress urinary incontinence, simultaneously the mesenchymal stem cells stimulate the regeneration of local tissues and blood vessels in a microenvironment through paracrine effect, enhance the function of the urethral sphincter, and after the filling agent and the allogeneic mesenchymal stem cells are gradually decomposed and absorbed by the body, the enhanced urethral sphincter plays a role in relieving female stress urinary incontinence for a long time.
Example 2:
an injection preparation for treating female stress urinary incontinence comprises mesenchymal stem cells and a filler; the volume ratio of the mesenchymal stem cells to the filler is 1: 2; the mesenchymal stem cells are derived from autologous endometrium, are prepared, amplified and cultured on a cell preparation platform conforming to GMP standard until 3 rd-5 th generation is frozen for standby, and are qualified in quality inspection; culture and harvestBy starvation therapy, inoculation density was 1 x 105And/ml, washing and replacing serum-free and phenol red-free DMEM culture solution when the growth density reaches about 75%, collecting supernatant after 24 hours, and measuring the growth factors by an enzyme-linked immunosorbent assay (ELISA) method as follows:
TABLE 1 determination of umbilical cord-derived mesenchymal Stem cell secretion growth factor Table (pg/ml)
| VEGF | bFGF | TGF | PDGF |
| Generation 3 1# | 215 | 42 | 119 | 7020 |
| Generation 3 # 2 | 528 | 21 | 106 | 4910 |
| Generation 5 1# | 103 | 97 | 206 | 5840 |
| 2# of 5 th generation | 117 | 32 | 212 | 5360 |
| DMEM control | 94 | 5 | 6 | 0 |
The filler is mainly sodium hyaluronate, the molecular weight of the filler is 100 ten thousand, and the filler is prepared into 1% concentration by using normal saline for later use;
taking umbilical cord mesenchymal stem cells 1 x 10 each time72ml in total, and preparing 2ml of medical grade sodium hyaluronate (molecular weight 100 ten thousand) with 1% concentration by using physiological saline, and mixing to obtain the medical grade sodium hyaluronate.
When in use, the injection needle is injected from the mucosa of the anterior wall of the vagina at a position 1.5cm away from the open end of the vagina, the needle head enters the gap between the connective tissues of the anterior wall of the vagina and the posterior wall of the middle section of the urethra, then 0.5-1ml of injection preparation is respectively pushed into the upper, the lower, the left and the right 4 points of a circle with the radius of 0.5cm by taking the needle inlet point as the center, and the injection preparation is pushed while retreating until a semi-surrounding bag is formed around the posterior wall of the middle section of the urethra;
wherein the number of mesenchymal stem cells in single injection dose is 1 × 107-5×107A plurality of; the injection preparation can be repeatedly injected, 2-3 times are injected in each course of treatment, the interval is 1-2 months, and the treatment amount of the 2 nd to 3 rd times is determined according to the symptom relieving condition of a patient.
Example 3:
an injection preparation for treating female stress urinary incontinence comprises mesenchymal stem cells and a filler; the volume ratio of the mesenchymal stem cells to the filler is 1: 3; the mesenchymal stem cells are derived from autologous fat, are prepared, amplified and cultured on a cell preparation platform conforming to GMP standard until 3 rd-5 th generation is frozen for standby, and are qualified in quality inspection;
the filler is mainly sodium hyaluronate, the molecular weight of the filler is 200 ten thousand, and the filler is prepared into 1% concentration by using normal saline for later use;
taking umbilical cord mesenchymal stem cells 1 x 10 each time72ml in total, and preparing 2ml of medical grade sodium hyaluronate (molecular weight 200 ten thousand) with 1% concentration by using physiological saline, and mixing to obtain the medical grade sodium hyaluronate.
Test example 1:
patient li XX, age 45 years;
the main complaints are: involuntary urine overflow in 7 years when the abdominal pressure is increased during laughing or sneezing
The past medical history: li XX, age 45, with menstrual disturbances and progressive reduction in menstrual flow with urinary incontinence (coughing and sneezing with increased abdominal pressure) for 7 years
Related family history: without special features
Gynecological examination: lax, dry and astringent vulva and vagina, bulging of the anterior wall, pressure test (+)
And (3) diagnosis: simple stress urinary incontinence in women
Treatment: suggested injection therapy (3 times a treatment course)
Clinical trials were conducted using the injection preparation prepared in example 1 of the present invention and the injection method in example 1, and the clinical effects were: urinary incontinence improved significantly about 1 week after the first treatment; disappearance of urinary incontinence symptoms after the second treatment; after 3 rd treatment, no recurrence was observed for two consecutive years.
Test example 2:
patient aged X, age 52 years;
the main complaints are: involuntary urine spill over 3 years during coughing and laughing or sneezing.
The past medical history: after menopause for 4 years, hot flashes, depression, emotional agitation and dry and astringent vagina appear.
Related family history: there is no special feature.
Gynecological examination: extreme laxity, dryness and bulging of the anterior wall of the vulva and vagina, pressure test (+).
And (3) diagnosis: simple stress urinary incontinence in women
Treatment: the injection therapy is recommended (2 times in one course).
Clinical tests were carried out using the injection preparation prepared in example 2 of the present invention and the injection method in example 2, and the clinical effects were: urinary incontinence was significantly improved about 10 days after the first treatment; the symptoms of urinary incontinence substantially disappeared after the second treatment, and the effect was continuously observed.
Test example 3:
the injection preparation prepared by the embodiment 1 of the invention and the injection method of the invention are adopted to observe the curative effect of dozens of volunteers, the cure rate of a patient can reach more than 90 percent within 2 years, the improvement rate can reach 100 percent, and a safe, effective, noninvasive, simple, convenient, feasible and no obvious side effect treatment method is provided for the middle-aged and the elderly women suffering from female stress urinary incontinence.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.