Preparation method of compound oral patch for oral mucosal epithelial cells

A technology of epithelial cells and oral mucosa, applied in the field of cell biology, can solve problems such as difficult to achieve therapeutic effect, loss of oral mucosa tissue, increased trauma, etc., to improve the efficiency of oral mucosa repair, good cell compatibility, and good antigen-free sexual effect

Inactive Publication Date: 2017-11-10
上海白衣缘生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Oral mucosa, as the first line of defense of the human body's biological barrier, plays a vital role in daily life, but the location of oral mucosa and its surrounding environment lead to a variety of factors that can cause loss of oral mucosal tissue, such as oral and maxillofacial surgery Tumor resection, alveolar surgery before trauma and repair often have large oral mucosal defects, which are often repaired clinically with skin grafts or flap transfers, but skin grafts often cause oral cavity stenosis and affect function due to contracture, and local flaps Metastasis often causes local swelling and poor shape, which is difficult for patients to accept
Moreover, autologous tissue transplantation is a treatment mode of "repairing damage by trauma" at the expense of normal tissue, which not only increases new trauma, but also increases the probability of surgical complications, and the limited source of autologous tissue limits its clinical application
In recent years, with the rise of biomaterials, clinical oral mucosal defect repair has achieved relatively satisfactory therapeutic effects, but for large-area oral mucosal defects, there are still major limitations, and it is difficult to achieve the expected therapeutic effect

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] 1. Preparation of oral patch: The oral patch was prepared using the submucosa of the small intestine. The method is to obtain the material from the submucosa of pig small intestine, and obtain it after decellularization, DNA depletion, virus inactivation, freeze-drying and irradiation sterilization.

[0025] 2. Primary culture of oral mucosal epithelial cells: Obtain oral mucosal tissue in the oral cavity of surgical patients under sterile conditions. Rinse repeatedly with double-antibody saline containing 100ug / ml penicillin and 100ug / ml streptomycin, then rinse with PBS solution, remove submucosal tissue with ophthalmic scissors, cut into small pieces of about 5mm×5mm, and put them in a petri dish Inside, add DKSFM medium containing 0.25% Dispase Ⅱ and submerge the tissue block, digest at 4°C for 16 hours, and separate the superficial epithelium and subepithelial layer with ophthalmic tweezers. The epithelial layer was digested with 0.25% trypsin at 37°C for 15 minut...

Embodiment 2

[0029] 1. Preparation of oral patch: The oral patch was prepared using the submucosa of the small intestine. The method is to obtain the material from the submucosa of pig small intestine, and obtain it after decellularization, DNA depletion, virus inactivation, freeze-drying and irradiation sterilization.

[0030]2. Primary culture of oral mucosal epithelial cells: Obtain oral mucosal tissue in the oral cavity of surgical patients under sterile conditions. Rinse repeatedly with double-antibody saline containing 100ug / ml penicillin and 100ug / ml streptomycin, then rinse with PBS solution, remove submucosal tissue with ophthalmic scissors, cut into small pieces of about 5mm×5mm, and put them in a petri dish Inside, add DKSFM medium containing 0.25% Dispase Ⅱ and submerge the tissue block, digest at 4°C for 17 hours, and separate the superficial epithelium and subepithelial layer with ophthalmic forceps. The epithelial layer was digested with 0.25% trypsin at 37°C for 15 minutes...

Embodiment 3

[0034] 1. Preparation of oral patch: The oral patch was prepared using the submucosa of the small intestine. The method is to obtain the material from the submucosa of pig small intestine, and obtain it after decellularization, DNA depletion, virus inactivation, freeze-drying and irradiation sterilization.

[0035] 2. Primary culture of oral mucosal epithelial cells: Obtain oral mucosal tissue in the oral cavity of surgical patients under sterile conditions. Rinse repeatedly with double-antibody saline containing 100ug / ml penicillin and 100ug / ml streptomycin, then rinse with PBS solution, remove submucosal tissue with ophthalmic scissors, cut into small pieces of about 5mm×5mm, and put them in a petri dish Inside, add DKSFM medium containing 0.25% Dispase Ⅱ and submerge the tissue block, digest at 4°C for 18 hours, and separate the superficial epithelium and subepithelial layer with ophthalmic forceps. The epithelial layer was digested with 0.25% trypsin at 37°C for 15 minute...

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PUM

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Abstract

The invention discloses a method for preparing an oral patch of composite oral mucosal epithelial cells, which comprises the following steps: preparing the oral patch by using the submucosa of the small intestine, cultivating the oral mucosal epithelial cells by a subculture method, and inoculating the oral mucosal epithelial cells into the oral cavity The patched cell culture dishes were cultured at 37°C in a 5% CO2 incubator. After the culture was terminated, the oral patch of composite oral mucosal epithelial cells was obtained. The preparation method of an oral patch of composite oral mucosal epithelial cells provided by the present invention has sufficient material sources, can make oral mucosal epithelial cells proliferate in large quantities in the oral patch, and provides a new method for the repair and reconstruction of large-area defects in the oral mucosa in clinical practice. way.

Description

technical field [0001] The invention relates to a method for preparing an oral patch made of composite oral mucosa epithelial cells, belonging to the field of cell biology technology. Background technique [0002] Oral mucosa, as the first line of defense of the human body's biological barrier, plays a vital role in daily life, but the location of the oral mucosa and its surrounding environment lead to a variety of factors that can cause loss of oral mucosal tissue, such as oral and maxillofacial surgery Tumor resection, trauma, and alveolar surgery before repair often have large oral mucosal defects, which are often repaired clinically by skin grafting or flap transfer, but skin grafting often causes oral cavity narrowing and affects function due to contracture, and local flaps Metastasis often causes local swelling and poor shape, which is difficult for patients to accept. Moreover, autologous tissue transplantation is a treatment mode of "repairing damage by trauma" at...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36A61L27/38A61L27/50C12N5/071
CPCA61L27/3629A61L27/3666A61L27/3687A61L27/3691A61L27/3813A61L27/3869A61L27/50A61L2300/412A61L2430/40C12N5/0632C12N2509/00
Inventor 李康乐汪文涛葛翠兰沈健峰韩韦红
Owner 上海白衣缘生物工程有限公司
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