Corneal graft

A graft and cornea technology, applied in the fields of medicine and biomedical engineering, can solve the problems of limited application, large damage to the donor cornea, and difficulty in obtaining a large number of viable seed cells.

Active Publication Date: 2007-07-18
上海组织工程研究与开发中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no doubt that autologous corneal stroma is the best choice for the carrier, but autologous corneal stroma is very precious to patients, while allogeneic corneal donors are very limited
Corneal stromal cells are difficult to obtain, causing great damage to the donor cornea, and it is difficult to obtain a large number of active seed cells, which limits its application

Method used

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preparation example Construction

[0055] The preparation method of the tissue engineered corneal stroma of the present invention is simple and convenient, and a certain amount of seed cells are mixed with pharmaceutically acceptable biodegradable materials.

[0056] The shape of the tissue-engineered corneal stromal graft of the present invention is not particularly limited. Typically, the shape of the graft is the same or substantially the same shape as the mammalian cornea. Taking a cornea-like graft as an example, its diameter is 3-5 mm and its thickness is 500-1000 μm.

[0057] The biodegradable material of the present invention is a solid material, and the seeding density is in ml of cell suspension. The seed cell concentration in the tissue engineered corneal stroma of the present invention is usually about 1×10 5 to 5×10 8 / ml or higher, preferably 1×10 6 to 1×10 8 / ml, more preferably 5×10 6 to 5×10 7 / ml. Usually, the concentration of the seed cell suspension is adjusted with the culture mediu...

Embodiment 1

[0066] Isolation and Culture of Rabbit Skin Fibroblasts

[0067] Pentobarbital sodium (30 mg / kg) was injected intravenously into the rabbit's ear margin. After the anesthesia was stable, the skin on the back of the rabbit was prepared. Routinely sterilize the drape, and take a full-thickness skin slice of about 5cm×3cm under sterile conditions. The subcutaneous adipose tissue of the skin graft was removed, and the dermis was reserved, which was washed in PBS. Then, cut the skin into about 1cm 3 The tissue pieces were placed in a 50ml centrifuge tube, 20ml of dispase II (purchased from Roche) was added, and digested at a constant temperature of 4°C for 16-24 hours. Then the epidermis was gently torn off, and the dermis tissue was placed in 0.2% collagenase (purchased from Gibco) for 2 hours at 37°C with constant temperature and shaking for digestion. The cell suspension was collected, centrifuged at 1500r / min for 5min, and discarded. Supernatant, washed with PBS repeatedly 3...

Embodiment 2

[0071] Preparation of bioscaffold polyglycolic acid (PGA)

[0072] After taking 2 mg of PGA with a diameter of 14 μm and shaping it (Figure 1), soak it in 75% ethanol for 30 minutes for disinfection, rinse it repeatedly with phosphate-buffered saline (PBS) for 3 times, dry it naturally, and place it in a 6-well plate for later use .

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Abstract

A cornea matrix transplant for providing proper carrier used for cornea transplantation is prepared from the pharmacologically acceptable biodegradable material and fibroblasts through mixing to inoculate said fibroblasts onto said biodegradable material.

Description

technical field [0001] The invention relates to the fields of medicine and biomedical engineering, and more specifically relates to using fibroblasts to construct corneal stroma grafts and its preparation method and application. Background technique [0002] Severe corneal disease can cause blindness, causing much inconvenience and suffering. Currently, corneal transplantation is the only reliable and effective treatment for clinical corneal blindness. The development of microsurgical techniques has greatly improved the success rate of corneal transplantation. However, due to the lack of corneal donors, there are still many patients who cannot receive keratoplasty. Moreover, postoperative immune response still exists in allogeneic corneal transplantation, and even donor rejection occurs, resulting in surgical failure. [0003] In the exploration of expanding the sources of corneal donors, researches on heterogeneous corneas and artificial corneal transplantation have been...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61F2/14A61L27/50A61L27/38A61L27/00A61F9/007
Inventor 曹谊林崔磊张艳青刘伟张文杰周广东胡晓洁杨光辉王会勇
Owner 上海组织工程研究与开发中心
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