Medical cornea paster and its preparation method

A patch and cornea technology, applied in the field of medicine, can solve the problems of limited source of materials, limited source of corneal limbus, limited effect, etc.

Inactive Publication Date: 2007-09-12
北京科宇联合干细胞生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] 1. Short-term corneal transparency after traditional keratoplasty, if the diagnosis is clear, it cannot be combined with surgery
Its main disadvantage is that only the central ф7mm optically clear area is transplanted, and it is ineffective for the loss of stem cells. Due to the obvious rejection, long-term use of immunosuppressants is required to cooperate with the treatment
[0005] 2. Although there is no immune rejection in autologous corneal limbus transplantation, the source of materials is limited. The main disadvantage is that it increases the risk of disease in healthy eyes. It cannot be transplanted in a 360° circle, and the effect is limited.
[0006] 3. Donor materials for allogeneic limbal transplantation are extremely limited, treatment opportunities are very few, and immune rejection is obvious, long-term medication is required, and the cost is high
Stem cells are the source of cell proliferation and differentiation in self-renewing tissues. Clinically, stem cells solve the problem of organ source caused by organ transplantation. Recently, corneal limbal stem cell transplantation has become a hot spot in the treatment of corneal diseases. The main method is to take autologous or allogeneic Limbal stem cells from the corneal limbus are planted on the amniotic membrane for transplantation to treat corneal diseases. However, due to the limited source of the limbal limbus, the rejection of allogeneic limbal stem cells is obvious after transplantation. Therefore, although it has a significant therapeutic effect, it has not been widely used in clinical practice.

Method used

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  • Medical cornea paster and its preparation method
  • Medical cornea paster and its preparation method
  • Medical cornea paster and its preparation method

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Experimental program
Comparison scheme
Effect test

Embodiment approach

[0021] 1. Raw materials:

[0022] (1) Medium a-MEM was purchased from GIBCO BRL (Grand Island, NY, USA), and its components include essential amino acids, proteins and polysaccharides for maintaining cell growth, product number: 1206692.

[0023] (2) Fetal bovine serum (FBS), purchased from Hyclone (Logan, UT), is an essential nutrient for cell growth, and its products are strictly in accordance with the quality standards of fetal bovine serum.

[0024] (3) Trypsin-EDTA, purchased from GIBCO BRL (Grand Island, NY, USA), is used for digesting cells.

[0025] (4) CD34, CD44, CD45, CD38, CD90, CD71, CD147, and HLA-DR were purchased from BD Company and are used to identify antibodies for markers of mesenchymal stem cells.

[0026] (5) Amniotic membrane: taken from the placenta of cesarean section women, preoperative detection of maternal serum HbsAg, anti-HCV, HCV-RNA, anti-HDV, anti-HEV, anti-HIV-1 / 2, HIV-1-RNA, RPR and CMV-DNA are negative, and the test method adopts the clini...

Embodiment 1

[0048] Ocular Surface Epithelial Recovery Evaluation

[0049] Total

[0050] Table 1: Corneal recovery of rats in different control groups 4 weeks after surgery

Embodiment 2

[0052] Restoration of Vision in Burned Rats

[0053] Through the tracking of the visual reflex state of the experimental rats by the optokinetic tracker, the total distance of motion and the trajectory of the single eye before the burn, the single eye before the burn, and the single eye after the burn are compared (see Fig. 6a, Fig. 6b, Fig. 6c, Fig. 6d, Fig. 6e, Fig. 6f), it can be seen that there is a significant difference in the tracking of the visual status of rats by the optomotor tracker, and the optomotor tracker works normally. Different types of cells were transplanted after the burn, and the data of each group were compared horizontally, and the P values ​​were compared through the optokinetic responses of different gratings (see Table 2, Table 3, Table 4, and Table 5, and the comparison between mesenchymal stem cells and other types of cells was obtained). The transplantation method can significantly improve the visual recovery effect of rats.

[0054] Th...

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Abstract

A medical cornea patch and its preparing method are disclosed, which implants autogenous mesenchyma stem cell to amnion for being carrier to treat cornea disease. The process includes: obtaining bone marrow 10ml from healthy person in clinical practice, diluting by PBS in ratio of 1:1, adding 5ml of Ficoll, centrifuging for 20 minutes, sucking out the nebulous single nuclei cell suspension, washing three times by PBS, implanting to 10% FBS containing culture medium of a-MEM, fetching 2X105 cell of the fourth generation to treated amnion with size of 1.5cmX1.5cm after 10 to 14 days, adding to culture medium to continue culturing, till the cell grow fully of the whole amnion.

Description

Technical field: [0001] The invention belongs to the technical field of medicine, and in particular relates to a patch prepared by planting mesenchymal stem cells on amnion for treating corneal diseases and a preparation method thereof. Background technique: [0002] The treatment of corneal injury is a complicated clinical problem. The cornea is divided into five layers, namely corneal epithelium, anterior boundary layer, corneal stroma layer, posterior boundary layer and corneal endothelial layer. Clinical corneal diseases include physical and chemical burns, mechanical trauma, infection, recurrent pterygium, Steven -Johnson's syndrome, tetanoid, trachoma, etc. Severe conjunctival and corneal diseases are still a major problem for ophthalmologists in the treatment of vision restoration. After corneal burns, a large number of limbal stem cells are lost, the corneal epithelium cannot be repaired normally, the cornea is cloudy, new blood vessels increase and form fibrosis, a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/50A61F2/14A61K48/00A61K9/70A61P27/02
Inventor 马艳玲李凌松
Owner 北京科宇联合干细胞生物技术有限公司
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