Air-dried amniotic membrane powder and application

An amniotic membrane, air-drying technology, applied in powder delivery, medical raw materials derived from mammals, drug combinations, etc., can solve problems such as disease transmission, inconvenience in use, and difficulty in sterilization

Inactive Publication Date: 2021-08-20
3 Cites 0 Cited by

AI-Extracted Technical Summary

Problems solved by technology

[0004] Glycerol preservation of amniotic membrane can fully preserve the histological structure and cytokine content of amniotic membrane, but it is difficult to sterilize, which may lead to disease transmission
[0005] Although freeze-dried amnion can retain growth factor activity better, its activity is still lower than that of fresh amnion, and the mechanical strength of freeze-dried amnion is weak. significant loss
[0006] Amniotic membrane extract is another hot spot in the research and development of new amniotic membrane products. It has a variety of high-level growth factors, but the amni...
View more


The invention discloses air-dried amniotic membrane powder and application thereof, relates to an amniotic membrane product, and particularly relates to a preparation method and application of amniotic membrane powder. The air-dried amniotic membrane powder is prepared by air-drying a fresh sterile and virus-free amniotic membrane at a constant temperature of 36-37 DEG C and grinding the amniotic membrane into the amniotic membrane powder. The air-dried amniotic membrane powder disclosed by the invention is simple and convenient to prepare, stable in property at room temperature and easy to store, and is a convenient and effective amnion product. The AM-FS daub prepared from amniotic membrane powder is prepared by loading amniotic membrane powder by using fibrin glue as a transfer matrix, contains abundant growth factors from amniotic membrane tissues, combines excellent biological properties of the fibrin glue, has an ocular surface tissue damage repairing effect similar to that of a fresh amniotic membrane, and overcomes the defectsthat the fresh amniotic membrane needs to be sutured, the surgical operation difficulty is high, and the operation fails due to easy falling, and the air-dried amniotic membrane powder is suitable for repairing various types of eye surface tissues and skin tissue injuries.

Application Domain

Powder deliverySenses disorder +3

Technology Topic

Tissue damageAnatomy +7


  • Air-dried amniotic membrane powder and application
  • Air-dried amniotic membrane powder and application
  • Air-dried amniotic membrane powder and application


  • Experimental program(2)

Example Embodiment

[0029] Example 1: Preparation method of wind dried sheepf powder, including the following steps:
[0030] S1, acquired amniovascular, select HIV, HBV, HCV, syphilis and other infectious disease tests, pregnant women, washed abandoned placenta within 24 hours of post-production, and dispelled off the amniotic membrane.
[0031] S2, wash the blood clots, mucus, mucus, washed with aseptic saline, and then placed in the physiological saline containing 50 ug / ml green-streptomycin, 100 ug / ml neomycin, 215 ug / ml amphotericin B repeated rinsing 45min Finally, it was flushed 3 times with aseptic saline.
[0032] S3, the wind-dried amniotic membrane, the washed amnion is flat, fixed on a support frame and then placed in a 37 ° C constant temperature circulation wind drying tank, stands for 2 h, and sufficiently removes the water-dried amniotic membrane.
[0033] S4, polishing powder, plasticized amniotic tissue after 37 ° C constant temperature drying into the stainless steel centrifuge tube specially equipped with a grinding nitrogen, ground 2 min, ground, 75 Hz, grind 180s, and obtain a diameter <260 um. Powder.
[0034] S5, sterilization, using 15-30 kGY γ ray irradiation, sterilized new wind-dried amniotic powder.
[0035] The amniotic powder obtained in this Example 1 was detected and compared to the expression of TGF-β, NGF R, HGF, EGF, and BFGF in the fresh amniocenter membrane. In addition to the expression of NGF R below the fresh amniotic membrane, the five cytokines is lower than that of the fresh amniotic membrane, and the amount of expression in the initial amniotic powder is higher than that of the fresh amniotic membrane group, and the difference has statistically significant (adj.p.val <0.05 And LOGFC> 0.263), TGFβ has no statistically significant significance in the initial amniotic membrane group (LOGFC <0.263), indicating that the new amniotic powder is preferred to retain the active ingredient in the amniotic tissue. The results are shown in Table 1.
[0036] Table 1: The difference in growth factors in new wind-dried amniotic powder and fresh amniotic membranes;
[0038] Note: There is statistically significant difference in P value (adj.p.val) <0.05, logfc> 0.263
[0039] The above data is selected by the raw data to normalize the Raybiotech software, and the analysis method is analyzed. The method is moderated T-Statistics, the packet is LIMMA, from R / Bioconductor; Filter the results of statistical significance Analysis.
[0040] Although the freeze-dried amniotic membrane can reserve the active cytokine, the content of various cytokines is lower than the fresh amniotic membrane. Therefore, the new wind-dried amniotic powder is directly compared with the fresh amniocentesis. The conclusion is five cytokines. In addition to NGF R, the amount of expression of the remaining cytokines in amniotic membranes is higher than that of the fresh amniotic membrane group, which can confirm the new type of air-dried amniotic powder has a living cytokine content similar to the fresh amniotic tissue.
[0041] The prepared amnimal powder was stored at room temperature, 4 ° C, and 30d, respectively, respectively, detected changes in the growth factor of three time points, respectively. figure 1 Indicated. The expression of EGF and NGF at 30 days decreased, and it was lower than the fresh amniotic membrane group. The growth factor of amniotic powder in the normal temperature storage group was higher than the -20 ° C storage group; the BFGF in the amniotic powder, HGF in various The expiration of the preservation condition and the preservation time group is higher than its expression of the fresh amniotic membrane group, and the expression difference between the growth factor of the amniotic powder in the room temperature storage group and the preservation group is not significant, so that the amaine powder is stable. Save at room temperature.

Example Embodiment

[0042] Example 2: Ams membrane-fibrin cement containing amniotic membranes (AM-FS: Amniotic MEMBRANE-FIBRIN Sealant), the preparation steps are as follows:
[0043] S1, the amnimeric powder prepared in Example 1 was thoroughly mixed with the catalyst solution in pig fibromy protein glue, and prepared according to mass ratio, and the amniotic powder concentration was 130 mg / ml, 65 mg / ml, 32 mg / ml, 16 mg / ml, 8 mg / ML, 4 mg / mL, 2 mg / ml, 1 mg / ml, 0.5 mg / ml, 0.25 mg / ml, 0 mg / ml of amniotic powder - catalyst mixture.
[0044] S2, sequentially take the above concentration of the amniotic powder-catalyst 50 ul, and the 24-well cell culture plate is added, and evenly placed on the bottom of the culture plate, and the equal volume main glue solution is sequentially added to the above culture holes, gently shock, mix well, formation According to the mass ratio, the amniotic powder concentration is: 65 mg / ml, 32 mg / ml, 16 mg / ml, 8 mg / ml, 4 mg / ml, 2 mg / ml, 1 mg / ml, 0.5mg / ml, 0.25 mg / ml, 0.125mg / ML, 0 mg / ml of AM-FS cement.
[0045] The cement of different concentrations of amniotic powder was selected from the 2nd generation of corneal epithelial cells (CEC: CORNEAL Epithelial Cells) to study in vitro cell experiments in vitro cell experiments, and studied the effects of CEC proliferation. Press 1 × 10 3 The CEC density of a / well was inoculated on a 24-well cell culture plate that has been paved in AM-FS cement, and 24-well cell culture plates were placed in 37 ° C, and the volume fraction was 5%. 2 Culture in the incubator for 72 h. The cells of each well were digested with 0.25% trypsin, and the cell suspension of each well was collected, 1000 r / min, centrifuged for 5 min, and the 96-well cell culture plate was added, and the cultured for 24 h was continued. 10 ul of CCK-8 solution was added to each well, and 2 h was continued to determine the absorbance (a) value at 450 nm. All 4 sub-holes per group were repeated 3 times.
[0046] According to CCK-8 results, the maximum AM-FS cement concentration of 0.25 mg / ml is the best ratio concentration, and the results are seen figure 2.
[0047] Am-fs cement concentration of amniotic powder was 0.25 mg / ml of AM-FS cement in the burn therapeutic experiment in rabbit. Building a rabbit severe eye oint base burn model, a total of 32 eyes; according to different treatments, model animals are randomly divided into 4 groups: AM-FS cement group 8 8 eyes, fresh amniotic membrane graft group 8 8 8 eyes, fibrin glue FS (FS is Fibrin Sealant abbreviation) group 8 8 eyes, 8 8 eyes in an antibiotic control group. The four sets of rabbits rows of rabbit row, compared, comparative cornea, and corneal neovascular displacement, comparative corneal. image 3 , Figure 4 Down:
[0048] At 28 days after treatment, there was no statistically significant difference between the AM-FS cement group corneal turbidity and the fresh amniotic membrane graft (P <0.05), and the two groups of corneal turbidity scores were lower than the antibiotic control group (P <0.05). The lower the corneal turbidity score, the more transparent the cornea, the AM-FS cement has the same effect of reducing the corneal turbidity after alkali burn, and the effect of restoring the corneal transparency is the same as the effect of the fresh amniotic membrane transplant. It is superior to the antibiotic control group.
[0049] After alkali burn increases with time, the AM-FS cement group, fresh amniotic membrane group, FS group, the newly ravron area is less than the antibiotic control group (P <0.05), although fresh amniotic membrane transplantation angle The new blood vessel area was less than the AM-FS cement group (P <0.05) in 7d, 21d, and 28d, but the difference was small (0.34, 0.38, 0.88), and the AM-FS cement was visible to the fresh amniotic membrane. Inhibition of the corneal neovascularization of the alkali burn, and the two methods are similar.
[0050] At the 21st, four groups of rabbits, tissue fixtures fixed eyeballs, compared the degree of corneal tissue new blood vessels and inflammatory response, Figure 5 Indicated. AM-Fs cement can alleviate corneal inflammatory response and neovascularization compared to FS group and antibiotic control group.
[0051] BRIEF DESCRIPTION OF THE DRAWINGS AM-FS gear can effectively promote the repair of rabbits with weight alkali burn. After AM-FS cement treatment, the recovery of corneal epithelial healing and transparency, inhibiting the effect of corneal neovascularization is better than antibiotic and simple fibrinogenic treatment group, which can achieve the same treatment with fresh amniotic membrane.
[0052] The wind-dried amniotic powder of the present invention is simple, stable in room temperature, easy to store, is a convenient and effective amniotic article. In this way, the amniotic powder in the amniotic membrane is small in the amniotic tissue, which can be compounded into a wound repair, a wound repair for minimally invasive intervention or special demand, can also be made with a variety of materials. Molecular polymers and synthetic polymer crosslinking are used in the field of tissue engineering, providing new ideas for the development of new amniotic membrane products.
[0053] The AM-FS cement prepared by amniotic powder is made of fibrin glue as a transfer matrix load amniotic membrane, which contains both the rich growth factor from the amniotic tissue, combining the fine biological properties of fibrin glue, and eye on the eyes The effect of forming damage is similar to the fresh amniot membrane. At the same time, it has overcome the fresh amniotic membrane needs to be stitched. The surgical operation is difficult, and it is easy to fall off, and the AM-FS cement can repeatedly drop the eye watch, form a uniform Soft gel-like substances have certain attraction and plasticity, no need to sew, easy to use, strong eye-catching, suitable for various types of eye-catching tissues and skin tissue damage.



Description & Claims & Application Information

We can also present the details of the Description, Claims and Application information to help users get a comprehensive understanding of the technical details of the patent, such as background art, summary of invention, brief description of drawings, description of embodiments, and other original content. On the other hand, users can also determine the specific scope of protection of the technology through the list of claims; as well as understand the changes in the life cycle of the technology with the presentation of the patent timeline. Login to view more.
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products