CN110393621A - A kind of fixed device of intrauterine device - Google Patents
A kind of fixed device of intrauterine device Download PDFInfo
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- CN110393621A CN110393621A CN201910549558.8A CN201910549558A CN110393621A CN 110393621 A CN110393621 A CN 110393621A CN 201910549558 A CN201910549558 A CN 201910549558A CN 110393621 A CN110393621 A CN 110393621A
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- alloy
- intrauterine device
- fixing
- intrauterine
- magnesium
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- 229910000861 Mg alloy Inorganic materials 0.000 claims abstract description 44
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- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 claims description 9
- 229910052725 zinc Inorganic materials 0.000 claims description 9
- 229910052761 rare earth metal Inorganic materials 0.000 claims description 6
- WHXSMMKQMYFTQS-UHFFFAOYSA-N Lithium Chemical compound [Li] WHXSMMKQMYFTQS-UHFFFAOYSA-N 0.000 claims description 4
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- WPBNNNQJVZRUHP-UHFFFAOYSA-L manganese(2+);methyl n-[[2-(methoxycarbonylcarbamothioylamino)phenyl]carbamothioyl]carbamate;n-[2-(sulfidocarbothioylamino)ethyl]carbamodithioate Chemical compound [Mn+2].[S-]C(=S)NCCNC([S-])=S.COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC WPBNNNQJVZRUHP-UHFFFAOYSA-L 0.000 claims description 4
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- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
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- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- KBMLJKBBKGNETC-UHFFFAOYSA-N magnesium manganese Chemical class [Mg].[Mn] KBMLJKBBKGNETC-UHFFFAOYSA-N 0.000 description 2
- SYJBLFMEUQWNFD-UHFFFAOYSA-N magnesium strontium Chemical class [Mg].[Sr] SYJBLFMEUQWNFD-UHFFFAOYSA-N 0.000 description 2
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- 208000036029 Uterine contractions during pregnancy Diseases 0.000 description 1
- GANNOFFDYMSBSZ-UHFFFAOYSA-N [AlH3].[Mg] Chemical class [AlH3].[Mg] GANNOFFDYMSBSZ-UHFFFAOYSA-N 0.000 description 1
- PGTXKIZLOWULDJ-UHFFFAOYSA-N [Mg].[Zn] Chemical class [Mg].[Zn] PGTXKIZLOWULDJ-UHFFFAOYSA-N 0.000 description 1
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- ZFXVRMSLJDYJCH-UHFFFAOYSA-N calcium magnesium Chemical class [Mg].[Ca] ZFXVRMSLJDYJCH-UHFFFAOYSA-N 0.000 description 1
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- 235000014655 lactic acid Nutrition 0.000 description 1
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- GCICAPWZNUIIDV-UHFFFAOYSA-N lithium magnesium Chemical class [Li].[Mg] GCICAPWZNUIIDV-UHFFFAOYSA-N 0.000 description 1
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- SJCKRGFTWFGHGZ-UHFFFAOYSA-N magnesium silver Chemical class [Mg].[Ag] SJCKRGFTWFGHGZ-UHFFFAOYSA-N 0.000 description 1
- QRNPTSGPQSOPQK-UHFFFAOYSA-N magnesium zirconium Chemical class [Mg].[Zr] QRNPTSGPQSOPQK-UHFFFAOYSA-N 0.000 description 1
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- 239000004626 polylactic acid Substances 0.000 description 1
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- XJMOSONTPMZWPB-UHFFFAOYSA-M propidium iodide Chemical compound [I-].[I-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CCC[N+](C)(CC)CC)=C1C1=CC=CC=C1 XJMOSONTPMZWPB-UHFFFAOYSA-M 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
- A61F6/00—Contraceptive devices; Pessaries; Applicators therefor
- A61F6/06—Contraceptive devices; Pessaries; Applicators therefor for use by females
- A61F6/14—Contraceptive devices; Pessaries; Applicators therefor for use by females intra-uterine type
Landscapes
- Health & Medical Sciences (AREA)
- Reproductive Health (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Heart & Thoracic Surgery (AREA)
- Vascular Medicine (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Orthopedics, Nursing, And Contraception (AREA)
Abstract
The present invention relates to intrauterine device fields, and in particular, to a kind of fixed device of intrauterine device.The fixed device of intrauterine device of the invention, including magnesium alloy fixed cone, magnesium alloy are made of Mg-X alloy or Mg-X-Y alloy.Present invention employs the magnesium alloys with excellent mechanical performance and degradation property as fixed device, increases intensity in intrauterine device fixation procedure；Reduce the problem of falling off and move down of intrauterine device, there is good biocompatibility and degradation property；Safely and effectively, structure designs advantages of simple, using simple, cost is relatively low.
Description
Technical Field
The invention relates to the field of intrauterine devices, in particular to an intrauterine device fixing device which is a magnesium alloy device for fixing an intrauterine device, has high fixing strength, good biocompatibility and simple operation, and can be degraded and absorbed by a human body.
Background
Fertility regulation is an important content of reproductive health, about 50 ten thousand women die each year worldwide due to pregnancy and childbirth syndrome, 25% -30% of the women die of involuntary pregnancy abortion, and the reduction of accidental pregnancy is a problem to be solved urgently. At present, China is still in the peak period of fertility, the number of delivery people reaches more than 1600 ten thousand per year, about 40 percent of the delivery people are cesarean sections, and the cesarean section yield of some hospitals is even as high as 60 to 80 percent. But due to the weak consciousness of postpartum contraception, the involuntary pregnancy rate and the artificial abortion rate within 1 year after birth are higher than the average level of women of childbearing age. The risk of artificial abortion in a short period after delivery is high, and an efficient contraceptive method should be considered preferentially.
WHO medical standards for contraceptive methods propose various contraceptive methods suitable for pregnant women WHO are born by caesarean section. Greater restrictions are placed on the choice of contraceptive drugs after delivery, taking into account whether or not lactation is taking place and the time of delivery; lactation amenorrhea contraception is only suitable for a few women, usually with a relatively high failure rate, and is also not suitable for long-term use. The IUD is placed in a safe, effective, simple, convenient, economic and reversible long-acting contraceptive measure, the IUD only acts on the local part of the uterus to play a contraceptive role, less interference is generated on an organism, the IUD is placed immediately after delivery and takes effect immediately, and the limitation of lactation is avoided (except for hormone-containing IUD), but the IUD has higher shedding rate due to larger uterine cavity, uterine mouth relaxation, uterine contraction and the like after delivery, and the application of the method is limited.
The only special IUD used for immediate postpartum placement at present is the Ginna IUD, namely the Ginni IUD and a degradable fixing cone are formed, wherein the fixing cone is a polylactic acid (PLA) high-molecular biodegradable polymer, is degraded into lactic acid and water in the uterine muscle wall for 2-3 months, and is discharged along with urine. The fixing system can effectively solve the problems of high dropping rate and low continuous rate of IUDs placed after abortion and postpartum. However, the defects of strong hygroscopicity, low thermal stability and poor flexibility and extensibility are found in the using process, the strength in the fixing process is not enough, and the processing into the special-shaped material is difficult to some extent.
In order to improve the fixing device, we consider using a new generation of degradable medical metal material magnesium alloy as the fixing device. Magnesium and magnesium alloy have excellent mechanical properties and degradability as a new-generation medical implant material, attract more and more attention, and have an important position in the field of biological materials. Although there are patent applications (for example, CN 102978493A, CN 102978495A, CN 109680195 a) related to magnesium alloy in the prior art, these applications are mostly directed to scaffolds or orthopedic devices in organs such as heart, etc., and the content of alloy elements other than Mg is too high, which is poor in biocompatibility with uterus, and is unfavorable for proliferation of endometrial epithelial cells, endometrial smooth muscle cells and endometrial stromal cells, and is not suitable for intrauterine application.
Disclosure of Invention
The invention aims to solve the technical problem that the prior intrauterine device is easy to fall off and move down in the uterus, and provides a magnesium alloy intrauterine device fixing device with excellent mechanical property and degradation property. The intrauterine device has good biocompatibility while solving the problems of the intrauterine device that the intrauterine device moves downwards and falls off.
In order to improve the fixing device, we consider using a new generation of degradable medical metal material magnesium alloy as the fixing device. Magnesium and magnesium alloy have excellent mechanical properties and degradability as a new-generation medical implant material, attract more and more attention, and have an important position in the field of biological materials.
The specific technical scheme of the invention is as follows:
the intrauterine device fixing device comprises a magnesium alloy fixing cone, wherein the magnesium alloy is made of Mg-X alloy or Mg-X-Y alloy; wherein,
the Mg-X alloy comprises 0-5% of X by mass and not 0% of X by mass, wherein the X is at least one of calcium, aluminum, strontium, zinc, manganese, lithium, zirconium, silver and rare earth elements; or,
the Mg-X-Y alloy may have X in a mass percent of 0% to 5%, and X in a mass percent of other than 0%, Y in a mass percent of 0% to 1%, and Y in a mass percent of other than 0%, for example, Y in a mass percent of but not limited to 0.1%, 0.2%, 0.5%, 0.8%, 1%, etc.;
x and Y are at least one of calcium, aluminum, strontium, zinc, manganese, lithium, zirconium, silver and rare earth elements, and are different elements.
The magnesium alloy used in the present invention includes, but is not limited to, magnesium-calcium series alloy, magnesium-aluminum series alloy, magnesium-strontium series alloy, magnesium-zinc series alloy, magnesium-rare earth series alloy, magnesium-manganese series alloy, magnesium-lithium series alloy, magnesium-zirconium series alloy, magnesium-manganese series alloy or magnesium-silver series alloy, or ternary series or multicomponent series magnesium alloy formed by combining these systems. In some embodiments, the magnesium alloy is a magnesium-rare earth alloy, in some embodiments, the magnesium alloy is a magnesium-tin-zinc-manganese alloy, and in some embodiments, the magnesium alloy is a magnesium-strontium alloy.
The intrauterine device fixing device according to the present invention, wherein preferably the Mg-X alloy has a mass percentage of X of 0.5% to 2%, for example, the mass percentage of X may be 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, etc.;
alternatively, the mass percentage of X in the Mg-X-Y alloy is 0.5% to 2%, such as but not limited to 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, and the like.
According to the intrauterine device fixing device of the present invention, it is further preferred that X in the Mg-X alloy is calcium or zinc, or X and Y in the Mg-X-Y alloy are calcium or zinc, respectively, and X and Y are different elements.
The magnesium alloy has biodegradability, and the magnesium alloy can basically meet the requirements of biomedical use, namely, the degradation amount of magnesium and other alloy elements in the degradation process of the magnesium alloy is controlled within a range of not causing toxic reaction of cells or tissues of an organism. The degradation rate of magnesium alloys depends on various factors, including the composition of the material itself, the surface state, the microstructure (grain size precipitates, etc.), the amount of impurities, the processing technique, etc. These factors are known to those skilled in the art and can be adjusted according to methods known in the art so that the release rate of the magnesium alloy can be controlled within a specifically desired range. For example, the degradation rate of the magnesium alloy can be controlled by controlling and adjusting the alloy proportion, the grain size, the impurity content and the processing technology of the material.
The preparation method of the magnesium alloy is not limited, any method capable of preparing the magnesium alloy can be adopted, and the method belongs to the conventional means in the field.
The specific shape of the magnesium alloy fixing cone is not particularly limited, and the magnesium alloy fixing cone can be implanted into or fixed on any shape of the inner wall of the uterus. Alternatively, the fixing cone may be a cone, an anchor, a hook, a spear, a rake or any other shape that can provide an anchoring effect, and has a knot that can tie the surgical thread to fasten, or clamp or clasp the surgical thread, so that the fixing cone and the surgical thread are tightly combined.
The intrauterine device fixing device according to the present invention further comprises a surgical wire, wherein one end of the surgical wire is fixed to the tail of the magnesium alloy fixing cone. The specific fixing method is not limited herein, and any fixing method in the field can be used. Such as, but not limited to, tying, or clamping, or clasping a knot of the surgical cable to tightly couple the fixation awl and the surgical cable.
Furthermore, the surgical thread may be made of any material that does not cause adhesion, such as but not limited to non-absorbable thread such as polypropylene, nylon, polyester, silk thread, etc., or absorbable thread such as polyglycolic acid, collagen, etc., as long as the complete absorption period is longer than the uterine recovery period.
The invention also provides an intrauterine device, which comprises an intrauterine device main body and the intrauterine device fixing device.
The intrauterine device according to the present invention, wherein the head of the intrauterine device body is fixedly connected to the other end of the operation wire. The specific fixing connection manner is not limited herein, and any fixing manner in the art may be adopted, such as, but not limited to, a metal buckle, a direct clamping or knotting manner, and the like.
The main body of the intrauterine device of the present invention is not particularly limited, and may be any intrauterine device in the art. For example, but not limited to, the intrauterine device body includes IUDs having various shapes such as T-shape, I-shape, intrauterine shape, gamma-shape, fancy, O-shape, etc.
The fixing device of the invention as a novel intrauterine device has the advantages that:
1. the magnesium alloy with excellent mechanical property and degradation property is adopted as a fixing device, so that the strength of the intrauterine device in the fixing process is increased;
2. the problems of falling off and downward movement of the intrauterine device are reduced, and the intrauterine device has good biocompatibility and degradation performance;
3. safe and effective, simple and reasonable in structural design, simple to use, lower in cost and the like.
Drawings
FIG. 1a is a schematic view of an intrauterine device fixing device (T-shaped IUD) of the present invention;
FIG. 1b is a schematic view of an intrauterine device fixing device (IUD type I) of the present invention;
FIG. 1c is a schematic view of an intrauterine device fixing device (IUD of uterine cavity type) of the present invention;
FIG. 2 shows the corrosion patterns of the surface of HP-Mg, Mg-0.8Ca and Mg-2Zn after 10 days of immersion in SUF, wherein (a) (b) (c) is HP-Mg material, (d) (e) (f) is Mg-0.8Ca material, (h) (i) (j) is Mg-2Zn material, (a) (d) (g) has pH6.0, (b) (e) (h) has pH7.0, and (c) (f) (i) has pH 7.9;
FIG. 3 is the EDS result corresponding to FIG. 2, with the alphabetical order, j-r corresponding one-to-one to A-I as shown by the boxes in (a) - (I);
FIG. 4 shows the corrosion patterns of the surface of HP-Mg, Mg-0.8Ca and Mg-2Zn after 42 days of immersion in SUF, wherein (a) (b) (c) is HP-Mg material, (d) (e) (f) is Mg-0.8Ca material, (h) (i) (j) is Mg-2Zn material, (a) (d) (g) has pH6.0, (b) (e) (h) has pH7.0, and (c) (f) (i) has pH 7.9;
FIG. 5 is the EDS result corresponding to FIG. 4, with the alphabetical order, j-r corresponding one-to-one to A-I as shown by the boxes in (a) - (I);
FIG. 6 shows the corrosion patterns of the surface of HP-Mg and Mg-2Zn after 120 days of immersion in SUF, wherein (a) (b) (c) is HP-Mg material, (d) (e) (f) is Mg-2Zn material, (a) (d) has pH6.0, (b) (e) has pH7.0, and (c) (f) has pH 7.9;
FIG. 7 is the EDS result corresponding to FIG. 6, g-l corresponding alphabetically to blocks A-F in (a) - (F);
FIG. 8 shows the morphological comparison of HP-Mg (a), Mg-0.8Ca (b) and Mg-2Zn (c) extracts with human endometrial epithelial cells cultured in negative control (d) (normal complete medium) and positive control (e) (10% DMSO addition).
Reference numerals
1. A magnesium alloy fixing cone 2, an operation line 3 and an intrauterine device main body.
Detailed Description
For better understanding of the technical solutions of the present invention, the following description is further provided with reference to the accompanying drawings.
The intrauterine device fixing device comprises a magnesium alloy fixing cone 1, one end of the magnesium alloy fixing cone 1 is fixed by an operation line 2 (polypropylene) with a knot, the intrauterine device also comprises an intrauterine device main body 3, the operation line 2 is connected with the intrauterine device main body 3, and the magnesium alloy fixing cone 1 is fixed in the uterine wall, so that the intrauterine device main body is not easy to move down and fall off.
Example 1
The structure of one embodiment of the invention is as follows: the fixing cone of the intrauterine device is made of Mg-Ca (the mass fraction of Ca is 1%) alloy and is connected with a T-shaped IUD (as shown in figure 1 a) through an operation line, the strength in the fixing process is increased, the Mg-1Ca alloy has no toxic effect on uterine cells, has good cell compatibility, has a certain effect of promoting the proliferation of the uterine cells, and can inhibit the expression of inflammatory factors.
Example 2
The second structure of the embodiment of the invention is as follows: the fixing cone is made of Mg-Zn (Zn mass fraction is 2%) alloy, and is connected with the I-type IUD (shown in figure 1 b) through an operation line, so that the strength in the fixing process is increased, and the Mg-Zn alloy has no toxic effect on uterine cells and has good cell compatibility.
Example 3
The third structure of the embodiment of the invention is as follows: the fixing cone is made of Mg-Ca (5% of Ca by mass) alloy, and is connected with the uterine cavity type IUD through an operation line (as shown in figure 1 c), the strength in the fixing process is increased, and the Mg-Ca alloy has no toxic effect on uterine cells and has good cell compatibility.
Example 4
The fourth structure of the embodiment of the invention is as follows: the fixing cone is made of Mg-Zn-Li (mass fraction of Zn is 1%, mass fraction of Li is 1%) alloy, and is connected with the uterine cavity type IUD through an operation line, so that the strength in the fixing process is increased, and the Mg-Zn-Li alloy has no toxic effect on uterine cells and has good cell compatibility.
Example 5
The fourth structure of the embodiment of the invention is as follows: the fixing cone is made of Mg-Ca-Zn (the mass fraction of Ca is 0.5 percent, the mass fraction of Zn is 0.5 percent) alloy, and is connected with the T-shaped IUD through an operation line, so that the strength in the fixing process is increased, and the Mg-Ca-Zn alloy has no toxic effect on uterine cells and has good cell compatibility.
Example 6 Corrosion Performance of a magnesium alloy fixation awl in simulated uterine cavity fluid
1. Preparation of simulated uterine cavity liquid
Electrolyte solution is selected to simulate uterine cavity solution (NaCl 4.97g/L, KCl 0.224g/L, CaCl)2 0.167g/L，NaHCO3 0.25g/L，Glucose 0.50g/L，NaH2PO4·2H2O0.072 g/L), the pH of SUF was adjusted to 6.0, 7.0 and 7.9 with hydrochloric acid and sodium hydroxide solution.
2. Preparation of materials
The experimental raw materials are pure magnesium with the purity of 99.99 wt%, pure calcium with the purity of 99.98 wt% and pure zinc with the purity of 99.99 wt%, and alloy components of HP-Mg (high-purity magnesium as a reference), Mg-0.8Ca (the mass percentage of Ca is 0.8%) and Mg-2Zn (the mass percentage of Zn is 2%) are selected and smelted. Smelting with SF6And CO2And (5) protecting the mixed gas. Heating the magnesium ingot to 650 ℃ to melt the magnesium ingot, adding the alloying element raw material preheated to 300 ℃, heating to 760 ℃ and 790 ℃, keeping the temperature and stirring for 30min, wherein the mold preheating temperature is 300 ℃, and the casting temperature is 710 ℃ and 740 ℃. Casting magnesium alloy ingotProcessing into cone with diameter of 3.5mm and height of 3.5mm, maintaining at 400 deg.C for 3 hr, and hot extruding. Extrusion ratio 17: 1, extrusion temperature 230 ℃. The experimental samples were then polished with 800 to 2000 particles of SiC paper. Thereafter, the cone was ultrasonically cleaned in anhydrous ethanol for 15 minutes, and then dried at room temperature.
3. Immersion test
The sample soak test was conducted according to ASTM-G31-72 with a solution volume to sample surface area ratio of 0.2mL/mm2The temperature is 37 +/-0.5 ℃. And taking out the sample after soaking for different time, washing the sample by deionized water, drying the sample at room temperature, observing the surface appearance change by a scanning electron microscope, and analyzing the element composition of the surface corrosion product by EDS. The method specifically comprises the following steps:
and taking out the samples at different time points of soaking, namely 10d, 42d and 120d respectively, drying at room temperature, and then placing the samples into a drying oven to be detected. After the soaking, the surface corrosion product was observed for surface corrosion morphology by scanning electron microscopy (S-4800 transmission scanning electron microscopy, HITACHI, Japan) and energy spectrum analysis (Quanta 200FEG, FEI).
4. Results of the experiment
FIGS. 2-3 show the topography of the material surface (FIG. 2) and EDS data (FIG. 3) after 10 days of immersion of HP-Mg, Mg-0.8Ca and Mg-2Zn in the SUF solution. SEM results show that the etch layer formed on the surface after 10 days immersion in the initial pH6.0 SUF was denser than those of the pH7.0 and pH7.9 groups, and the underlying layer under the etch layer was clearly visible under a high power mirror. And corrosion products on the Mg-0.8Ca and Mg-2Zn surfaces are more than on the HP-Mg surfaces. Elemental analysis of corrosion products by EDS showed that the elements contained were mainly C, O, Mg, P, Ca.
FIGS. 4-5 show the topography of the material surface (FIG. 4) and EDS data (FIG. 5) after immersion of HP-Mg, Mg-0.8Ca and Mg-2Zn in the SUF solution for 42 days. The results show that more corrosion products are deposited on the surface of the material than at 10 days. Whereas Mg-0.8Ca and Mg-2Zn form more corrosion products than HP-Mg surface material. The corrosion precipitation of HP-Mg surface corrosion products increases significantly with increasing pH. Elemental analysis of corrosion products by EDS showed that the elements contained were mainly C, O, Mg, P, Ca.
FIGS. 6-7 show the corrosion morphology (FIG. 6) and EDS data (FIG. 7) on the material surface after immersion of HP-Mg and Mg-2Zn in SUF solution for 120 d. At the moment, Mg-0.8Ca is corroded too fast, and is degraded into powder after being soaked for 120 days and completely disintegrated, so that the EDS data and the surface morphology of the powder cannot be obtained. SEM results are given for only the other two materials. The Mg-2Zn surface is observed to form a denser corrosion layer through SEM, and compared with 42d, the corrosion product formed on the surface presents a more compact ice flower shape. Whereas loose spherical corrosion products were visible on the HP-Mg surface. Elemental analysis of corrosion products by EDS showed that the elements contained were mainly C, O, Mg, P, Ca.
Example 7 evaluation of compatibility of magnesium alloy fixed cone cells
1. Preparation of materials
The same as in example 6.
2. Preparation of leaching liquor
The sample polished to # 2000 was washed in acetone, ethanol and deionized water for 10 minutes, dried at room temperature, and sterilized with ethylene oxide. Placing the sample in a sterile culture dish, and controlling the ratio of the leaching liquor to the sample surface area to be 1cm2Adding complete culture solution of human endometrial epithelial cells (DMEM-F12 complete culture medium) at a ratio of/mL, placing at 37 deg.C, 95% relative humidity, and 5% CO2Centrifuging twice at 1000rpm for 24 hr to remove alloy precipitate, measuring pH value and element concentration of the leaching solution, sealing, and storing in a refrigerator at 4 deg.C for no more than 1 week.
3. Flow cytometry determination of cell cycle of endometrial stromal cells
1) Cell cycle synchronization: the cells were cultured for 24h in serum-free medium, thereby synchronizing the cell cycle.
2) Culturing cells by using a leaching solution: sucking out serum-free culture medium, adding the prepared leaching solution, placing at 37 deg.C, 95% relative humidity, and 5% CO2The culture box is used for culturing for 24 hours.
3) Cell collection: the cell culture fluid was first collected into a 15mL centrifuge tube for use. And (3) digesting the cells for 2-3mins by 0.25% of pancreatin until the cells can be blown down by a pipette or a gun head, adding the previously collected cell sap, blowing down all adherent cells, and blowing off the cells. And collected again in the centrifuge tube.
4) Centrifuging: centrifuging at 1000g for 3-5mins, and precipitating cells. If the precipitation is insufficient, the Linchenne time can be prolonged properly or the centrifugal force can be increased slightly. The supernatant was freshly aspirated from the cells, and about 50. mu.L of the culture medium was left to prevent aspiration of the cells.
5) Resuspending: approximately 1mL of ice-cooled PBS was added, the cells were resuspended, and transferred to a 1.5mL centrifuge tube. The cells were again pelleted by centrifugation and the supernatant carefully aspirated, leaving approximately 50. mu.l of PBS to avoid aspiration of the cells. Gently flick the bottom of the centrifuge tube to properly disperse the cells and avoid cell clumping.
6) Cell fixation: adding into 1mL ice bath precooled 70% ethanol, lightly blowing and uniformly mixing, and fixing at 4 ℃ for 2h or more than 48 h. The effect is better after the fixation for 12-24 h. Centrifuging at 1000g for 3-5mins, and precipitating cells. If the precipitation is insufficient, the centrifugation time may be appropriately prolonged or the centrifugal force may be slightly increased. The supernatant was carefully removed and 50. mu.L of 70% ethanol remained to avoid cell aspiration. 1mL of ice-cooled PBS was added and the cells were resuspended. The cells were pelleted again by centrifugation and the supernatant carefully aspirated, 50. mu.LPBS could remain to avoid aspiration of the cells. Gently flick the bottom of the centrifuge tube to properly disperse the cells and avoid cell clumping.
7) Preparing an propidium iodide color solution:
1 sample
8) Dyeing: 0.5mL of propidium iodide staining solution was added to each tube of cell sample, and the cell pellet was slowly and thoroughly resuspended and incubated at 37 ℃ for 30mins in the dark. The cells can then be stored at 4 ℃ or in ice baths protected from light. After the staining is finished, the detection is preferably finished within 24h, and the flow detection is preferably finished on the same day.
9) Flow detection and analysis: the red fluorescence was detected with a flow cytometer at the 488nm excitation wavelength, with the light scattering detected. Cellular DNA content analysis and spectral scattering analysis were performed using software.
4. Results of the experiment
FIG. 8 shows the change in cell morphology after 5 days of incubation of endometrial epithelial cells with three materials of leaching solution. From the results, it can be seen that the negative control group had smooth cell edges and uniform growth, and exhibited the morphology of typical epithelial cells. While the positive control cells were fragmented and the cells were disrupted and dead. From the three materials, the growth state of the Mg-0.8Ca leaching solution cultured cells is similar to that of the negative control group, and the Mg-2Zn group has a small amount of cell death.
As can be seen from Table 1, compared with the blank control group, the proportion of G0/G1 phase of HP-Mg, Mg-0.8Ca and Mg-2Zn groups is reduced, the proportion of cells in S phase and G2/M phase is increased, but most of the cells are in G0/G1 phase, which shows that the stroma cells are still normal diploid after being acted with the HP-Mg, Mg-0.8Ca and Mg-2Zn materials, no heteroploids are generated, and the fact that the materials are not tumorigenic and carcinogenic is proved. The increase of the proportion of the cells in the S phase and the G2/M phase indicates that the material can increase the DNA synthesis of the endometrial stroma cells, promote the division and proliferation of the endometrial stroma cells and is beneficial to the growth of the endometrial stroma cells and the repair of endometrium in the menstrual cycle. Particularly, the S phase of Mg-0.8Ca is obviously higher than that of a control group (p is less than 0.05), which indicates that Mg-0.8Ca can obviously promote the proliferation of endometrial stromal cells.
TABLE 1 Effect of HP-Mg, Mg-0.8Ca and Mg-2Zn leach liquors on cell cycle of human endometrial stromal cells cultured for 24h
Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention and are not limited. Although the present invention has been described in detail with reference to the embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (9)
1. An intrauterine device fixing device, characterized in that it comprises a magnesium alloy fixing cone, the magnesium alloy is made of Mg-X alloy or Mg-X-Y alloy;
wherein the Mg-X alloy contains 0-5% of X by mass and not 0% of X by mass, and the X is at least one of calcium, aluminum, strontium, zinc, manganese, lithium, zirconium, silver and rare earth elements; or,
the Mg-X-Y alloy comprises, by mass, 0-5% of X, 0-1% of Y and 0-1% of Y, wherein X and Y are at least one of calcium, aluminum, strontium, zinc, manganese, lithium, zirconium, silver and rare earth elements, and are different elements.
2. The intrauterine device fixture according to claim 1, wherein the Mg-X alloy has X in the amount of 0.5-2% by mass, or the Mg-X-Y alloy has X in the amount of 0.5-2% by mass.
3. The intrauterine device fixing device according to claim 1 or 2, wherein X in the Mg-X alloy is calcium or zinc, or X and Y in the Mg-X-Y alloy are calcium or zinc, respectively, and X and Y are different elements.
4. The intrauterine device fixing device according to any of claims 1 or 2, wherein the magnesium alloy fixing cone is a cone, an anchor, a hook, a spear or a rake.
5. The intrauterine device fixing device according to claim 3, wherein the magnesium alloy fixing cone is a cone, an anchor type, a fishhook type, a spearhead type or a rake head type.
6. The intrauterine device fixing device according to any one of claims 1-3, 5, further comprising a surgical wire, one end of which is fixed to the tail of the magnesium alloy fixing cone.
7. The intrauterine device fixing device according to claim 4, further comprising a surgical wire, one end of which is fixed to the tail of the magnesium alloy fixing cone.
8. An intrauterine device comprising an intrauterine device body, characterized in that it further comprises an intrauterine device fixing device according to claim 6 or 7.
9. The intrauterine device according to claim 8, wherein the head of the intrauterine device body is fixedly connected with the other end of the operation wire.
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