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Decellularized scaffold material for regeneration of salivary gland and preparation method thereof

A technology of decellularized scaffolds and cell scaffolds, applied in tissue regeneration, medical science, prostheses, etc., can solve problems such as lack of salivary gland regeneration, achieve the effects of reducing chewing, improving quality of life, and improving saliva secretion

Pending Publication Date: 2018-05-29
BEIJING STOMATOLOGY HOSPITAL CAPITAL MEDICAL UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the prior art shows that this decellularized organ scaffold material is widely used in the research of heart, kidney, lung, pancreas and other organs, there is no report on salivary gland regeneration based on it so far.

Method used

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  • Decellularized scaffold material for regeneration of salivary gland and preparation method thereof
  • Decellularized scaffold material for regeneration of salivary gland and preparation method thereof
  • Decellularized scaffold material for regeneration of salivary gland and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1: Submandibular Gland Decellularization Treatment

[0044]

[0045] (1) Take 8-week-old male rats, after intraperitoneal anesthesia with chloral hydrate injection, T-shape neck incision to expose bilateral submandibular glands, remove the capsule, cut the submandibular glands and keep the submandibular gland ducts as much as possible; immerse in 0.01mol / L heparinized PBS (phosphate buffer saline, heparin concentration 25 μg / ml), shaking on a shaker for 15 minutes to remove blood;

[0046] (2) Add 12.5% ​​SDS (sodium dodecyl sulfate) aqueous solution of 1000 times the volume of the sample, put it in a shaker, shake and cultivate at room temperature at 200 rpm for 32 hours, and replace the SDS solution every 8 hours;

[0047] (3) Add sterile deionized water and shake for 4 hours to remove residual SDS;

[0048] (4) Add 1% Triton X-100 (polyethylene glycol octyl phenyl ether), and culture for 2 hours;

[0049] (5) Add 0.02mg / ml of DNAaseI (deoxyribonuclease I)...

Embodiment 2

[0060] Example 2: Isolation of Rat Submandibular Gland Cells

[0061] (1) Take 3-4 week old male rats, after intraperitoneal injection of anesthesia, peel off the capsule after incision of the neck, separate and extract the submandibular gland, and cut the submandibular gland tissue into pieces in the sterile operating table;

[0062] (2) Place the above tissue in the digestion solution containing 3mg / ml type I collagenase and 4mg / ml Dispase (neutral proteolytic enzyme), digest at 37°C for 1 hour, and shake constantly during the period. After the digestion was completed, the cells were collected through a 70 μm cell sieve, centrifuged at 1200 rpm for 5 min, and the supernatant was discarded and resuspended with MEGM medium to form a single cell suspension.

Embodiment 3

[0063] Example 3: Recellularization and three-dimensional culture of decellularized submandibular gland

[0064] (1) Slowly inject the single cell suspension described in Example 2 into the decellularized scaffold material through the submandibular gland duct with a 1ml syringe to complete the recellularization process;

[0065] (2) Place the cell-scaffold complex statically in the MEGM culture medium, and put it into an incubator to cultivate for 12 hours, so as to facilitate cell adhesion;

[0066] (3) Finally, put the cell-scaffold complex into a 50ml rotary three-dimensional organ culture system, add MEGM medium, adjust the rotation speed to 10rpm to ensure that the cell-scaffold complex is in a suspended state, and place it at 37°C, 5% CO 2 in the incubator. The medium was changed every 2 days, and the culture time was 14 days.

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PUM

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Abstract

The invention relates to a decellularized scaffold material for regeneration of salivary gland and a preparation method thereof. The preparation method comprises the following steps: (1) performing decellularized treatment on submaxillary gland of male rats; (2) performing cell separation of the submaxillary gland of the male rats; (3) performing re-cellularization and three-dimensional culture onthe decellularized submandibular gland. The decellularized scaffold material prepared by the method disclosed by the invention has the advantages of organ specificity and no immunological rejection reaction, and wide applicability to the regeneration of the salivary gland such as parotid gland, submandibular gland and submaxillary gland; the decellularized submaxillary gland as a scaffold material is renewedly transplanted into allogeneic SD (Sprague Dawley) rats and can mediate the regeneration of the salivary gland.

Description

technical field [0001] The invention belongs to the field of regenerative medicine, and in particular relates to a decellularized scaffold material for salivary gland regeneration and a preparation method thereof. Background technique [0002] Saliva is an important body fluid in the human body, mainly composed of three pairs of major salivary glands (parotid gland, submandibular gland, and sublingual gland, whose secretion accounts for about 90% of the total saliva) and a large number of minor salivary glands distributed under the oral mucosa Secretion formed. Salivary glands play an important role in maintaining normal physiological homeostasis of the oral cavity and upper gastrointestinal tract. Various reasons such as radiation injury, Sjogren's syndrome, and age-related changes can cause hypofunction of the salivary glands, which can cause a decrease in saliva secretion, resulting in obvious dry mouth, which can cause difficulties in chewing, swallowing, and speaking. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36
CPCA61L27/3633A61L27/3687A61L2430/00
Inventor 高振华王凌霄王松灵单兆臣李钧
Owner BEIJING STOMATOLOGY HOSPITAL CAPITAL MEDICAL UNIV
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